Weinman; Department of Internal Medicine, University of Kansas Medical Center, Kansas City, KS Alcoholic hepatitis is a severe form of alcohol-induced liver inflammation that occurs onl
Trang 1♦ Denotes AASLD Presidential Poster of Distinction
1194
Clinical study of liver regenerative therapy of cirrhosis
using autologous adipose tissue-derived stromal cells
and the characterization of the obtained stromal cells.
Yoshio Sakai 1 , Akihiro Seki 1 , Hajime Sunagozaka 1 , Takeshi
Ter-ashima 1 , Kazunori Kawaguchi 1 , Hatsune Mochida 2 , Alessandro
Nasti 2 , Geraldine B Buffa 1 , Masatoshi Yamato 2 , Kosuke Ishida 4 ,
Masaaki Takamura 2 , Soichiro Usui 2 , Takashi Wada 3 , Masao
Honda 1 , Shuichi Kaneko 4,1 ; 1 Department of Gastroenterology,
Kanazawa University, Kanazawa, Japan; 2 Disease Control and
Homeostasis, Kanazawa University, Kanazawa, Japan; 3
Depart-ment of Nephrology, Kanazawa University, Kanazawa, Japan;
4 System Biology, Kanazawa University, Kanazawa, Japan
Adipose tissue is enriched with mesenchymal stromal/stem cells
We conducted the clinical study for liver regenerative therapy
of cirrhosis by intrahepatic arterial administration of freshly
iso-lated autologous adipose tissue derived stromal/stem
(regener-ative) cells (ADRCs) (UMIN000009122, NCT01062750) We
also characterized the obtained fresh cells (ADRCs) as well as
the cultured expanded stromal cells (ADSCs) [Methods] The
objectives were cirrhosis patients who provided the written
informed consent The patients underwent liposuction of their
subcutaneous adipose tissues in their abdomen or buttock The
obtained adipose tissues were immediately processed using the
adipose-tissue dissociation equipment (Celution®, Cytori
Thera-peutics Inc ) to obtain autologous ADRCs The designated
num-ber of cells (3 3x10^5/Kg (n=2), 6 6x10^5/Kg (n=2)) were
infused through the catheter positioned at the hepatic artery
Safety evaluation was assessed 1 month after the treatment
Surface antigen of ADRCs and cultured cells (ADSCs) were
assessed by FACS Serum cytokine/chemokine concentration
was measured by Bio-Plex® We also analyzed gene
expres-sion of the freshly isolated ADRCs compared to the cultured
ADSCs by DNA microarray [Result] Four liver cirrhosis patients
were enrolled, (HI-01(type C), HI-03 (type C), HI-04 (NASH),
HI-05(type B)) The number of infused ADRCs were 2 2x10^7
~ 4 4x10^7 Among ADRCs and ADSCs, 10 3 ~45 8 %
and 80 998 9% of cells, respectively, expressed the
mesen-chymal stem cell surface marker CD44 No severe adverse
events occurred Three among 4 treated patients improved
serum albumin concentration during 36 months after treatment
Serum HGF, M-CSF, MIF, IL18, and IL-6 were elevated in all 4
patients one day after treatment The surplus ADRCs after
treat-ment were successfully expanded and the spindle-like shape of
mesenchymal stem cell was observed Gene expression profile
analysis using clusters analysis showed the two distinct
clus-ters discerning ADRCs from ADSCs, completely, regardless of
the patient’s etiologies of cirrhosis The freshly isolated ADRCs
were shown to involve inflammatory features, suggesting that
they were related to immunomodulatory biological effects
[Conclusion] Intrahepatic arterial administration of autologous
freshly isolated ADRCs with immunomodulatory biological
effects were confirmed to be safely conducted without serious
adverse events
Disclosures:
Shuichi Kaneko - Grant/Research Support: MDS, Co , Inc, Chugai Pharma , Co ,
Inc, Toray Co , Inc, Daiichi Sankyo , Co , Inc, Dainippon Sumitomo, Co , Inc,
Ajinomoto Co , Inc, Bristol Myers Squibb , Inc, Pfizer , Co , Inc, Astellas , Inc,
Takeda , Co , Inc, Otsuka„ÄÄPharmaceutical, Co , Inc, Eizai Co , Inc, Bayer
Japan, Eli lilly Japan
The following people have nothing to disclose: Yoshio Sakai, Akihiro Seki, Hajime
Sunagozaka, Takeshi Terashima, Kazunori Kawaguchi, Hatsune Mochida,
Ales-sandro Nasti, Geraldine B Buffa, Masatoshi Yamato, Kosuke Ishida, Masaaki
Takamura, Soichiro Usui, Takashi Wada, Masao Honda
1195 ♦
Therapeutic targeting of Hsp90 alleviates liver phage-specific NLRP3 inflammasome activation in alco- holic liver injury
macro-Daniel Bullock, Asmita Choudhury, Pranoti Mandrekar; Medicine, University of Massachusetts Medical Center, Worcester, MA
Background/Aims: Inflammasomes are multimeric protein plexes that respond to PAMPs or DAMPs that serve as a scaf-fold to promote cleavage of pro-caspase-1 and subsequent active IL-1β and IL-18 production Previous studies show activa-tion of NLRP3 inflammasome in alcoholic liver disease (ALD) Further the role of stress-mediated protein heat shock protein
com-90 (hspcom-90) as a chaperone for NLRP3 inflammasome scaffold was identified Studies in our laboratory have demonstrated that chronic alcohol induces hsp90 and its inhibition allevi-ates alcoholic liver injury Here we hypothesize that hsp90 is required for alcohol mediated NLRP3 inflammasome activity
in the liver, specifically in macrophages/Kupffer cells and its therapeutic targeting reduces active IL-1β in the alcoholic liver Methods: C57/BL/6J mice were subjected to 2 weeks and 6 weeks of chronic-single binge or chronic-multiple binge alco-hol feeding respectively Specific hsp90 inhibitor, 17-DMAG was injected intraperitoneally either every alternate day (5mg/kg) or at the end of the alcohol feeding (50mg/kg) Whole livers were harvested and subjected to inflammasome qPCR and IL-1β ELISA Hepatocytes and liver macrophages were iso-lated at the end of the feeding and NLRP3 and IL-1β analyzed For mechanistic studies, bone marrow derived macrophages (BMDM) were differentiated in vitro and stimulated with LPS
± ATP and DMAG (0 5μM-2 0μM) or directly heat shocked and stimulated with ATP, and analyzed for inflammasome mRNA and protein Results: Chronic alcohol mediated induc-tion of NLRP3 expression (p<0 002) was observed in alcohol exposed whole livers and restricted to isolated liver macro-phages/Kupffer cells, but not hepatocytes Inhibition of hsp90, using 17-DMAG significantly inhibited NLRP3 (p<0 01), ASC (p<0 01) and pro-caspase-1 (p<0 002) in liver macrophages exposed to chronic alcohol in vivo Bioactivity of 17-DMAG
in the liver was confirmed by induction of hsp70 expression Further 17-DMAG decreased chronic alcohol-binge induced IL-1β protein in whole livers (p<0 001) In vitro studies reveal that 17-DMAG and heat shock (induction of hsp70 indepen-dent of hsp90 inhibition) reduced active IL-1β (p<0 0002) and caspase-1 cleavage in BMDM, demonstrating that both inhi-bition of hsp90 and induction of hsp70 can prevent robust NLRP3 inflammasome signaling Conclusion: Our results sup-port our hypothesis that hsp90 is crucial in alcohol mediated NLRP3 inflammasome activation which is restricted to liver mac-rophages Overall we demonstrate clinical relevance of hsp90 inhibition in preventing NLRP3 inflammasome and active IL-1β production, and predict protective function of hsp70/hspA1A
in ALD Disclosures:
The following people have nothing to disclose: Daniel Bullock, Asmita hury, Pranoti Mandrekar
Trang 2Mechanisms that Control Cyp4a10 and Cyp4a14
Expression by Nuclear Receptor SHP and Rev-Erbα in
Alcoholic Fatty Liver
Zhihong Yang 1,2 , Yi Huang 1 , Will Bogaert 1 , Li Wang 1,2 ; 1
Uni-versity of Connecticut, Storrs, CT; 2 Veterans Affairs Connecticut
Healthcare System, West Haven, CT
Background & aims: Extensive alcohol consumption results in
alcoholic fatty liver (ALF) disease and alcoholic hepatitis This
study aims at identifying new regulators of ALF Methods: The
ethanol-binge model (NIAAA model) was used In short, two
month-old C57BL/6 (WT) and nuclear receptor Shp-/- mice
were fed with the Lieber-DeCarli diet containing 5% ethanol
ad libitum or pair-fed with an isocaloric control diet for 10
days On day 11, mice were oral-gavaged with a single dose
of 5g of ethanol (EtOH) or maltose dextrin (CTRL) per kg of
body weight After an original period of nine hours, the serum
and liver samples were then collected every six hours for the
following twenty-four hours The samples were subjected to
RNA-seq analysis Other methods include: qPCR, Western blot,
serum and liver lipid analysis, dual-luciferase reporter assay,
AAV8 virus transduction and shRNA knockdown Results: ALF
induced by ethanol-binge in WT mice were diminished in Shp
-/- mice, as determined by oil-red O and H&E staining, and
analysis of serum and liver lipid profile RNA-seq revealed a
group of cycling transcripts that were highly increased in WT
liver fed with ethanol but not in Shp-/- mice, including Cyp4a10
and Cyp4a14 Alcohol also induced Cyp4a10 and Cyp4a14
expression in WT but not Shp-/- MEFs Both Cyp4a10 and
Cyp4a14 expression showed a circadian rhythmic change
in WT mice fed with control liquid diet, and their rhythm was
shifted after the ethanol feeding and by overexpression of
Shp Luciferase reporter assay revealed that Cyp4a10 and
Cyp4a14 promoter activities were inhibited by Rev-Erbα, and
Shp repressed the inhibition of Erbα Knockdown of
Rev-Erbα by AAV8-shRNA increased Cyp4a10 and Cyp4a14
expression, whereas overexpressing Rev-Erbα repressed
Cyp4a10 and Cyp4a14 in mouse liver In addition,
Rev-Erbα agonist SR9009 blocked the induction of Cyp4a10 and
Cyp4a14 by ethanol Conclusions: The activation of Cyp4a10
and Cyp4a14 by ethanol may contribute to ALF in WT mice
The expression of Cyp4a10 and Cyp4a14 is controlled by
circadian clock regulators Rev-Erbα and Shp
Disclosures:
The following people have nothing to disclose: Zhihong Yang, Yi Huang, Will
Bogaert, Li Wang
1197
FOXO3-dependent, M1-selective hepatic macrophage
apoptosis is a normal mechanism that allows the liver to
tolerate alcohol with minimal or no inflammation
Zhuan Li, Jie Zhao, Steven A Weinman; Department of Internal
Medicine, University of Kansas Medical Center, Kansas City, KS
Alcoholic hepatitis is a severe form of alcohol-induced liver
inflammation that occurs only infrequently after heavy drinking
and almost never in the setting of a previously normal liver
This suggests that the liver has mechanisms that normally
pre-vent alcohol-induced inflammation We have previously shown
that alcohol treatment converts the transcription factor FOXO3
into a apoptotic factor and this somehow serves to
pro-tect the liver from alcohol-induced inflammation In this study
we aimed to examine the mechanisms of FOXO3-mediated
liver protection Methods: Wild-type (wt) and FOXO3-/- mice
were administered ethanol either with a single gavage (5g/
kg) or by feeding a Lieber-DeCarli ethanol-containing diet (5% vol/vol) for 10 days In some cases mice received a single dose of LPS (10μg/mouse) via intraperitoneal injection Liver and serum samples were collected and analyzed by histol-
ogy, RT-PCR, TUNEL staining, and ELISA assays Results: Acute
ethanol gavage to wt mice resulted in massive intrahepatic macrophage apoptosis as evidenced by TUNEL positivity and 50% decreases of macrophage marker F4/80 and CD68 mRNA This macrophage loss was entirely FOXO3-dependent and did not occur in FOXO3-/- mice Macrophage apoptosis was type specific Acute alcohol gavage only decreased M1 specific markers but had no effect on M2 markers We next
examined mouse peritoneal macrophages in vitro INF-γ
dif-ferentiated macrophages (M1) from wt mice underwent tosis in response to LPS FOXO3-/- M1 macrophages and IL-4 differentiated (M2) macrophages from either wt or knock out mice did not undergo apoptosis after LPS treatment Finally
apop-we examined the effects of chronic ethanol feeding on mice Ten-day feeding had no effect on hepatic macrophage num-ber or M1/M2 ratio in wt mice but increased the M1/M2 ratio in FOXO3-/- mice After 10 days of alcohol feeding we treated wt and FOXO3-/- mice with LPS The alcohol exposure produced an anti-inflammatory effect in wt mice with reduced LPS-induced production of pro-inflammatory cytokines (such as TNF-a, IL-6 and IL-1b) and no change in ALT In FOXO3-/- mice, the same alcohol exposure produced a pro-inflammatory effect with increased production of TNF-a, IL-6, IL-1b and a 5-fold
increase in ALT in response to the same LPS challenge
Conclu-sions: FOXO3-dependent, M1-specific macrophage apoptosis
is a novel mechanism that allows the intrahepatic macrophage population to change and become more anti-inflammatory in response to alcohol exposure This helps to generate hepatic alcohol tolerance Defects in FOXO3-depdendent apoptosis block this adaptation and sensitize the liver to inflammatory injury in response to alcohol
A Brenner 1 , Bernd Schnabl 1 ; 1 University of California San Diego,
La Jolla, CA; 2 University of Nebraska Medical Center, Omaha, NE; 3 J Craig Venter Institute, Rockville, MD; 4 NGM Biopharma- ceuticals, Inc., South San Francisco, CA; 5 The Salk Institute, La Jolla, CA
Background: Alcoholic liver disease (ALD) is associated with intestinal dysbiosis The aim of this study was to identify a mechanism of how changes in the intestinal microbiota con-tribute to ALD Methods and Results: Metagenomic sequencing
of intestinal contents demonstrated that chronic ethanol ing in mice is associated with an overrepresentation of bacte-rial genomic DNA encoding choloylglycine hydrolase, which deconjugates intestinal bile acids Using a Lieber-DeCarli alco-hol feeding model for 8 weeks, targeted metabolomics con-firmed an increased amount of unconjugated bile acids in the small intestine Unconjugated bile acids are rapidly absorbed
feed-by nonionic diffusion in the jejunum Mediated feed-by a lower FXR activity in enterocytes of the terminal ileum, lower FGF15 protein secretion resulted in increased hepatic Cyp7a1 protein
Trang 3♦ Denotes AASLD Presidential Poster of Distinction
expression and increased bile acid synthesis Total plasma and
hepatic bile acid levels were elevated after chronic ethanol
feeding Depletion of the commensal microbiota with
non-ab-sorbable antibiotics, decreased hepatic Cyp7a1 expression
and reduced alcoholic liver disease in mice, suggesting that
increased bile acid synthesis is dependent on gut bacteria To
restore enterohepatic circulation of bile acids, we used a
phar-macological intervention with the intestine-specific FXR agonist
fexaramine, which is not absorbed after oral gavage and does
not show any systemic FXR stimulation Daily gavage of
fex-aramine reduced hepatic Cyp7a1 protein expression following
chronic ethanol feeding Fexaramine treatment protected mice
from ethanol-induced liver injury, characterized by reduced
plasma ALT, hepatic triglycerides and inflammation As a
sec-ond precision target approach, we counterbalanced the effects
of reduced FGF15 with an adeno-associated virus (AAV)
expressing the human non-tumorigenic FGF19-variant V97 in
mice AAV expressing GFP was used as control A prevention
(AAV was injected 2 weeks prior to ethanol feeding) and
inter-vention trial (AAV was injected 3 weeks after ethanol feeding
was started) was performed In both trials, overexpression of
FGF19 reduced hepatic Cyp7a1 expression and
ethanol-in-duced liver disease Absorption and hepatic metabolism of
ethanol were not affected by either of our approaches
Conclu-sion: Chronic ethanol feeding causes an overexpression of bile
acid deconjugating enzymes in the intestinal microbiota,
result-ing in significant alterations of bile acid profiles
Precision-tar-geted interventions with the intestinal FXR-agonist fexaramine
and the FGF19-variant V97 restored bile acid homeostasis and
reduced ethanol-induced liver disease in mice
Disclosures:
Lei Ling - Employment: NGM Biopharmaceuticals, Inc ; Stock Shareholder: NGM
Biopharmaceuticals, Inc
Stephen J Rossi - Employment: NGM Biopharmaceuticals, Inc; Stock
Share-holder: NGM Biopharmaceuticals, Inc
Alex M DePaoli - Employment: NGM Biopharmaceuticals
Michael Downes - Consulting: metacrine
The following people have nothing to disclose: Phillipp Hartmann, Angela
Hor-vath, Peng Chen, Caroline T Seebauer, Ana Cristina Llorente-Izquierdo, Lirui
Wang, Yazen Alnouti, Derrick E Fouts, Ruth T Yu, Ronald M Evans, David A
Brenner, Bernd Schnabl
1199
Osteopontin ablation drives hematopoietic stem cell
mobilization and increases hepatic iron contributing to
alcoholic liver disease
Fernando Magdaleno 1 , Xiaodong Ge 1 , Holger Fey 2 , Chuck C
Blajszczak 1 , M Isabel Fiel 3 , Costica Aloman 2 , Natalia Nieto 1,2 ;
1 Pathology, University of Illinois at Chicago, Chicago, IL; 2
Med-icine, University of Illinois at Chicago, Chicago, IL; 3 Pathology,
Icahn School of Medicine at Mount Sinai, New York, NY
Background & Aim: Although it has been shown that
hema-topoietic stem cells (HSCs) restore injured parenchyma, the
continuous mobilization of bone marrow-derived HSCs into the
liver could contribute to alcoholic liver disease (ALD) To date,
the mechanisms underlying HSCs mobilization from the bone
marrow (BM) to the liver and its contribution to ALD remain
poorly defined Osteopontin (OPN) is a matricellular protein
and a negative regulator of HSC proliferation that controls
HSC lodgment and retention Thus, the aim of this study was
to investigate the role of OPN in restricting HSCs mobilization
into the liver and its contribution to ALD Methods: We
ana-lyzed young (14-16 wks ) and old (>1 5 yrs ) wild-type (WT)
littermates and global Opn knockout (Opn-/-) mice for the
pres-ence of BM mobilization into the liver In addition, 10 wks old
WT and Opn -/- mice were chronically fed with either the control
or the ethanol Lieber-DeCarli diet for 7 wks BM, blood, spleen and liver were analyzed by polychromatic flow cytometry for HSC progenitors and neutrophils Chemokines, growth factors and cytokines were measured in serum and liver H&E staining and scoring, Prussian blue staining for ferric iron deposits and Naphthol AS-D chloroacetate esterase staining for neutrophils
were performed in liver sections Results: Myeloid progenitor
cells, CD34+ and CD127+ hematopoietic cells were lower in
the BM from young compared to old Opn-/- mice, which played significant hepatosplenomegaly compared to their age-
dis-matched WT littermates Opn-/- mice had significantly elevated
Kitl mRNA levels albeit similar expression of Cxcr4 and Sdf-1α
mRNAs Hepatic chemokine (C-X-C motif) ligand 1 (CXCL1), macrophage inflammatory protein 1 alpha (MIP1α) and chemokine (C-C motif) ligand 5 (CCL5) as well as granulocyte colony-stimulating factor (G-CSF) and macrophage colony-stim-
ulating factor (M-CSF) were increased in Opn-/- mice ing potential engraftment of HSCs from the BM into the liver
suggest-Furthermore, Opn-/- mice had reduced transferrin receptor (TfR) expression accompanied with increased Epo and decreased Hamp mRNA expression, which are central players in iron homeostasis and iron uptake Indeed, Opn-/- mice showed iron deposits in the liver and in the BM but were absent in WT
littermates Ethanol-fed Opn-/- mice showed significant hepatic neutrophil infiltration and hemosiderin compared to WT mice
As a result, ethanol feeding caused greater liver injury in Opn /- compared to WT mice Conclusion: Opn ablation promotes
-HSC mobilization, neutrophil infiltration and iron deposits in the liver; therefore, enhancing the severity of ALD
Disclosures:
The following people have nothing to disclose: Fernando Magdaleno, Xiaodong
Ge, Holger Fey, Chuck C Blajszczak, M Isabel Fiel, Costica Aloman, Natalia Nieto
3 Division of Gastroenterology and Hepatology, Stanford sity School of Medicine, Stanford, CA; 4 Biostatistics, Brown Uni- versity School of Public Health, Providence, RI; 5 Transplantation, California Pacific Medical Center, San Francisco, CA; 6 Medicine, University of Illinois College of Medicine, Chicago, IL; 7 Medicine, University of Tennessee Health Science Center, Memphis, TN
Univer-Background and Aims: Alcoholic liver disease (ALD), the ond leading indication for liver transplantation (LT) worldwide,
sec-is facing a resurgence in the United States (U S ) We tigated the temporal trends and outcomes of ALD-related LT
inves-in the U S Methods: We conducted a retrospective cohort study utilizing the United Network for Organ Sharing registry from 2003 to 2014 to evaluate the frequency of ALD-related
LT Etiology-specific survival following LT, with a focus on the top three indications for LT, was evaluated with Kaplan Meier methods and multivariate Cox proportional hazards models Results: Overall, 63,061 adult patients underwent LT from 2003-2014, including 9,470 ALD (15 02%), 20,782 hepatitis
C virus (HCV) (32 96%), and 8,266 nonalcoholic titis (NASH) (13 11%) From 2003 to 2014, the number of LT secondary to ALD increased by 55%, whereas LT secondary
steatohepa-to HCV increased by 33% Among the steatohepa-top three indications for LT, the lowest 1-year, 3-year and 5-year post-transplant sur-vival was noted among patients with HCV The highest 1-year,
Trang 43-year, and 5-year post-transplant survival was noted among
patients with ALD (1-year = 90 95%; 95% CI, 90 38–91 53;
P < 001 vs HCV; 3-year = 85 37%; 95% CI, 84 66–86 08;
P < 001 vs HCV; 5-year = 81 82%; 95% CI, 81 05–82 61;
P < 001 vs HCV) On multivariate Cox proportional hazards
modeling, patients with ALD had significantly higher
post-trans-plant survival compared to HCV patients (HR, 0 80; 95% CI,
0 76–0 84; P < 001) Conclusion: Among the top indications
for LT, ALD has the highest long-term survival following LT ALD
is rising rapidly as an indication for LT and may, once again,
overtake NASH as the second leading etiology for LT in the
U S
Disclosures:
Satheesh Nair - Advisory Committees or Review Panels: Jansen; Grant/Research
Support: Gilead; Speaking and Teaching: Abbvie, Valeant, BMS, Intercept
Aijaz Ahmed - Consulting: Bristol-Myers Squibb, Gilead Sciences Inc , Roche,
AbbVie, Shire, Janssen pharmaceuticals; Grant/Research Support: Gilead
Sci-ences Inc
The following people have nothing to disclose: George Cholankeril, Ryan B
Perumpail, Menghan Hu, Avin Aggarwal, Andy Liu, Eric R Yoo
1201
Alcoholic hepatitis patients have a defect in
FOXO3-de-pendent myeloid cell apoptosis due to high SIRT
expres-sion
Zhuan Li, Brian Bridges, Jody C Olson, Steven A Weinman;
Department of Internal Medicine, University of Kansas Medical
Center, Kansas City, KS
Alcoholic hepatitis results from a failure to downregulate an
inflammatory response induced by alcohol We have
previ-ously shown that FOXO3 serves as an alcohol protection factor
by virtue of its ability to induce apoptosis of pro-inflammatory
macrophages This FOXO3-dependent apoptosis, which is
selective for M1 over M2 macrophages, requires the
JNK-de-pendent phosphorylation of FOXO3 at S574 and is induced
by LPS It has previously been shown that FOXO3 acetylation
also promotes its apoptotic activity but the relationship between
S574 phosphorylation and acetylation has not been examined
The aim of this study was to determine the relationship between
acetylation and phosphorylation in FOXO3-induced apoptosis
and determine whether defects in this system are present in
patients with acute alcoholic hepatitis Methods: THP-1 cells
were treated with LPS (100 ng/mL), a SIRT activator
(resvera-trol, 100 μM) or a SIRT inhibitor (CAY10591, 50 μM) FOXO3
acetylation and phosphorylation, and cell death were ated by immunoprecipitation, western blot and TUNEL assays Monocytes were isolated from human blood by density gradi-ent centrifugation They were treated with LPS for 24 hours in the absence or presence of a SIRT inhibitor (Sirtinol, 50 μM) and cell death was evaluated by caspase 3/7 activity and cell
evalu-viability Results: LPS treatment of THP-1 cells caused
JNK-de-pendent S574-FOXO3 phosphorylation and apoptosis LPS also induced hyperacetylation of FOXO3 which resulted from decrease in SIRT1 and SIRT7 Prevention of FOXO3 acetyla-tion by either SIRT activation or SIRT1 or SIRT7 overexpression completely blocked p-S574-FOXO3 formation and apoptosis Conversely, increasing FOXO3 acetylation by SIRT inhibition increased p-S574-FOXO3 and apoptosis A mutant FOXO3 lacking four known acetylation sites (K242, 259, 290, 569R) was not able to bind JNK and failed to induce apoptosis Pri-mary human peripheral blood monocytes (PBMs) isolated from healthy controls had low SIRT1 and SIRT7 and readily under-went FOXO3 phosphorylation and apoptosis; PBMs from alco-holic hepatitis patients had high expression of SIRT1 and SIRT7 and failed to phosphorylate FOXO3 and undergo apoptosis in response to LPS Apoptosis of these cells was restored by inhibi-
tion of SIRT activity Conclusions: FOXO3 acetylation regulates
monocyte apoptosis and this plays a role in downregulating the inflammatory response to alcohol High SIRT1 and SIRT7 levels in monocytes from patients with acute alcoholic hepatitis prevent this anti-inflammatory apoptosis response Loss of this FOXO3-dependent apoptosis pathway may thus contribute to the occurrence of alcoholic hepatitis
Ethanol-In-Chunki Kim 1 , Xin Shi 2 , Yoonhee Han 1 , Hong Shen 1,3 , Alvin asuria 1 , Jiayou Wang 1 , Jiashin Wu 1 , Ping Zhang 2 , Min You 1 ;
Jog-1 Pharmaceutical Sciences, Northeast Ohio Medical University, Rootstown, OH; 2 Integrative Medical Sciences, Northeast Ohio Medical University, Rootstown, OH; 3 Liver Diseases, Guangdong Hospital of Traditional Chinese Medicine at Zhuhai, Zhuhai, China
Abnormality of hepatic sirtuin 1 (SIRT1) plays a pivotal role
in the pathogenesis of alcoholic liver disease (ALD) In the present study, we investigated the role of myeloid cell-specific SIRT1 in the development of alcoholic steatohepatitis utilizing
a myeloid cell-specific SIRT1 knockout (mSIRT1KO) mouse model Matched mSIRT1KO mice and littermate control (WT) mice were pair-fed with either an ethanol containing or control diet using the Gao-binge ethanol feeding protocol Myeloid cell-specific SIRT1 deficiency exacerbated alcoholic liver injury
as demonstrated by aggravated inflammation, increased atosis, and elevated serum AST and ALT in mice after the alco-hol challenge Further analysis revealed that hepatic activity
ste-of myeloperoxidase (MPO) and gene expression ste-of Ly6G (a neutrophil marker) were mildly elevated in the ethanol-fed WT mice compared to WT controls However, the increases in MPO activity and Ly6G expression were markedly augmented
in the livers of ethanol-fed mSIRT1KO mice Flow cytometric analysis confirmed that the percentage of circulating and liver neutrophils bearing Gr1+/CD11b+ markers were elevated in the ethanol-fed WT mice compared to pair-fed WT controls Strikingly, myeloid cell-specific SIRT1 deficiency significantly augmented the percentage of neutrophils in the blood and the
Trang 5♦ Denotes AASLD Presidential Poster of Distinction
livers of ethanol-fed mice In line with this, mRNA expression
of the neutrophil-recruiting chemokines, including CCL2
(MCP-1), CCL3 (MIP-1α), CCL4 (MIP-1β), Cxcl2, and Cxcl10, were
robustly increased in the ethanol-fed mSIRT1KO mice, but not
in ethanol-fed WT mice Myeloid cell-specific SIRT1 deficiency
also markedly increased mRNA abundance of the
endothe-lium-specific molecules, including SELE, SELP, and ICAM-1,
whereas ethanol feeding to WT mice slightly increased mRNA
expression of those molecules compared to WT controls
Inter-estingly, gene expression of a macrophage marker F4/80+ in
liver was substantially decreased in both WT and mSIRT1KO
mice after ethanol administration, suggesting that ethanol
feed-ing might induce apoptosis of F4/80+-positive Kupffer cells
in these mice Collectively, our findings suggest that myeloid
cell-specific SIRT1 deficiency exaggerates the ethanol-induced
neutrophilic inflammation, which contributes to the
develop-ment of alcoholic steatohepatitis in mice Hence, myeloid
cell-specific SIRT1 may be a potential target for developing
effective therapy to treat human alcoholic steatohepatitis
Disclosures:
The following people have nothing to disclose: Chunki Kim, Xin Shi, Yoonhee
Han, Hong Shen, Alvin Jogasuria, Jiayou Wang, Jiashin Wu, Ping Zhang, Min
You
1203
Corticosteroids are the only remaining pharmacological
option for severe alcoholic hepatitis: a meta-analysis of
individual data on 1974 patients
Mark R Thursz 2 , Alexandre Louvet 1 , Dong Joon Kim 10 , Julien
Labreuche 9 , Stephen R Atkinson 2 , Sandeep S Sidhu 3 , John G
O’Grady 4 , Robert L Carithers 7 , Marie-Jose Ramond 11 , Charles
L Mendenhall 5 , Willis C Maddrey 6 , Timothy R Morgan 8 , Alain
Duhamel 9 , Philippe Mathurin 1 ; 1 Maladies de l’appareil digestif,
Hôpital Huriez, Lille, France; 2 Imperial College, London, United
Kingdom; 3 Dayanad Medical College, Ludhiana, India; 4 Institute
of Liver Diseases, London, United Kingdom; 5 Veteran Affairs,
Cincinnati, OH; 6 University of Texas, Dallas, TX; 7 University of
Washington, Seattle, WA; 8 Veteran Affairs, Long Beach, CA;
9 Departement de biostatistiques, Lille, France; 10 Hallym Medical
Center, Chuncheon, Korea (the Republic of); 11 Hôpital Beaujon,
Clichy, France
We performed a meta-analysis using individual patient data
from nine randomized controlled trials (RCTs) which evaluated
corticosteroids (CS), pentoxifylline (PTX) or their combination in
patients with severe alcoholic hepatitis (Maddrey discriminant
function (DF) ≥32) Aims: compare 28-day survival according
to treatment and analyze therapeutic response (Lille model)
Methods: We obtained individual data from 1974 patients,
originating from 9 RCTs, and performed 3 different
meta-anal-yses: CS vs placebo or control treatment (CTL) (n=6 trials),
CS vs PTX (n=2) and CS+PTX vs CS+placebo (n=3) Results:
First analysis: A total of 495 (CS) and 469 (CTL) patients were
included We found no heterogeneity between the trials (test
of interaction not significant: p=0 2) After stratification on
the trial, the HR of mortality at 28 days was 0 643 (95%CI:
0 482-0 855, p=0 002) in CS-treated patients as compared
to CTL After adjustment on DF, creatinine, age and
albu-min, the HR remained significant (HR=0 504, 95%CI 0
.367-0 693, p<.367-0 367-0.367-0.367-01) At day 7, there were more responders
(Lille model<0 45) in CS-treated patients as compared to
CTL (67 4 vs 54 1%, p<0 0001) and the OR of response in
CS-treated patients after adjustment on the trial as compared
to CTL was 1 749 (1 279-2 392, p=0 0005) On available
data at day 28, CS-treated patients had a higher
improve-ment in bilirubin from baseline than CTL: -9 7±0 6 vs -6 9±0 7
mg/dl, p=0 002 Second analysis: 335 (PTX) and 333 (CS) patients were included, without heterogeneity between the tri-als (p=0 3) HR of death at 28 days was 0 643 (0 439-0 943, p=0 002) in CS patients as compared to PTX and was still sig-nificant after adjustment on the same 4 variables (see above):
HR 0 655, p=0 04 There were more responders in CS patients
as compared to PTX: 67 3 vs 48 3%, p<0 0001 and OR for response in CS patients as compared to PTX was 2 29 (1 57-
3 35, p<0 0001) Third analysis: 441 (CS+PTX) and 445 (CS) patients were included, without heterogeneity between trials (p=0 96) After stratification on the trial, the HR of mortality at
28 days was not significant for the combination of CS+PTX as compared to CS alone: HR 0 9, 95%CI 0 63-1 29, p=0 58 The proportion of responders was the same in the two groups:
60 4 vs 64 8% respectively, p=0 25 Thus, OR for response
in CS+PTX patients was not significant (0 835, 95%CI 0
.61-1 61-15, p=0 3) Conclusion: Corticosteroids improve 28-day
sur-vival in patients with severe alcoholic hepatitis with a higher response rate as compared to PTX and CTL This treatment benefit is sustained until the end of therapeutic period The combination of CS and PTX does not add any additional effect Disclosures:
Mark R Thursz - Advisory Committees or Review Panels: Gilead, BMS, Abbott Laboratories, CN-Bio, Altimmune
Timothy R Morgan - Grant/Research Support: Merck, Abbvie, Genentech, ead, Bristol Myers Squibb
Gil-Philippe Mathurin - Board Membership: MSD, Janssen-Cilag, BMS, Gilead, Abvie, Verlyx; Consulting: Roche, Bayer
The following people have nothing to disclose: Alexandre Louvet, Dong Joon Kim, Julien Labreuche, Stephen R Atkinson, Sandeep S Sidhu, John G O’Grady, Robert L Carithers, Marie-Jose Ramond, Charles L Mendenhall, Willis C Mad- drey, Alain Duhamel
1204
Assessment of nutritional status through bioelectrical impedance is a key determinant of outcome in alcoholic hepatitis: a prospective study
Laurent Spahr 1 , Laurence Genton 2 , Laura Rubbia-Brandt 3 , sandra Oropesa 1 , Emiliano G Giostra 1 ; 1 Gastroenterology, University Hospital of Geneva, Geneva, Switzerland; 2 Nutrition, University Hospitals of Geneva, Geneva, Switzerland; 3 Clinical pathology, University Hospitals of Geneva, Geneva, Switzerland
Cas-Alcoholic hepatitis (AH) is a severe complication of alcoholic liver disease with an elevated mortality rate on the short term
in spite of corticosteroids A concomitant poor nutritional status may impact prognosis Aim : We explored the specific role of malnutrition in a prospective cohort of patients with regards to clinical outcome at 6 months Methods : a group of 45 patients admitted to hospital with AH (age 52 9 yrs ; MELD score 19 9 + 5 3 ; Child-Pugh 10 4 + 1 3 ; BMI 26 1 kg/m2 ; C-reactive protein (CRP) 18 8 + 14 9 mg/l ; 62% treated with steroids) were evaluated at baseline using arm muscle circumference (AMC, anthropometric measurement) and phase angle value
by bioelectrical impedance (BIA) This analysis is based on the conductivity properties of body tissues, is performed at bedside, and is a sensitive and non invasive mean to determine nutritional status Clinical condition and alcohol relapse were carefuly monitored Results : Prevalence of malnutrition was 60% (by AMC, < 10th percentile) and 74% (by BIA), statisti-cally higher (p<0 05) as compared to a group of excessive drinkers without AH (n=20) During follow-up, the mortality was 23% Alcohol relapse occurred in 28% of patients Multivari-ate Cox regression analysis showed that phase angle by BIA was an independent predictor of poor outcome at 6 months (p=0 05, 95%CI [0 26-0 99] Other variables such as AMC (p=0 07), age (p=0 83), BMI (p=0 34), MELD score (p=0 35),
Trang 6livers of ethanol-fed mice In line with this, mRNA expression
of the neutrophil-recruiting chemokines, including CCL2
(MCP-1), CCL3 (MIP-1α), CCL4 (MIP-1β), Cxcl2, and Cxcl10, were
robustly increased in the ethanol-fed mSIRT1KO mice, but not
in ethanol-fed WT mice Myeloid cell-specific SIRT1 deficiency
also markedly increased mRNA abundance of the
endothe-lium-specific molecules, including SELE, SELP, and ICAM-1,
whereas ethanol feeding to WT mice slightly increased mRNA
expression of those molecules compared to WT controls
Inter-estingly, gene expression of a macrophage marker F4/80+ in
liver was substantially decreased in both WT and mSIRT1KO
mice after ethanol administration, suggesting that ethanol
feed-ing might induce apoptosis of F4/80+-positive Kupffer cells
in these mice Collectively, our findings suggest that myeloid
cell-specific SIRT1 deficiency exaggerates the ethanol-induced
neutrophilic inflammation, which contributes to the
develop-ment of alcoholic steatohepatitis in mice Hence, myeloid
cell-specific SIRT1 may be a potential target for developing
effective therapy to treat human alcoholic steatohepatitis
Disclosures:
The following people have nothing to disclose: Chunki Kim, Xin Shi, Yoonhee
Han, Hong Shen, Alvin Jogasuria, Jiayou Wang, Jiashin Wu, Ping Zhang, Min
You
1203
Corticosteroids are the only remaining pharmacological
option for severe alcoholic hepatitis: a meta-analysis of
individual data on 1974 patients
Mark R Thursz 2 , Alexandre Louvet 1 , Dong Joon Kim 10 , Julien
Labreuche 9 , Stephen R Atkinson 2 , Sandeep S Sidhu 3 , John G
O’Grady 4 , Robert L Carithers 7 , Marie-Jose Ramond 11 , Charles
L Mendenhall 5 , Willis C Maddrey 6 , Timothy R Morgan 8 , Alain
Duhamel 9 , Philippe Mathurin 1 ; 1 Maladies de l’appareil digestif,
Hôpital Huriez, Lille, France; 2 Imperial College, London, United
Kingdom; 3 Dayanad Medical College, Ludhiana, India; 4 Institute
of Liver Diseases, London, United Kingdom; 5 Veteran Affairs,
Cincinnati, OH; 6 University of Texas, Dallas, TX; 7 University of
Washington, Seattle, WA; 8 Veteran Affairs, Long Beach, CA;
9 Departement de biostatistiques, Lille, France; 10 Hallym Medical
Center, Chuncheon, Korea (the Republic of); 11 Hôpital Beaujon,
Clichy, France
We performed a meta-analysis using individual patient data
from nine randomized controlled trials (RCTs) which evaluated
corticosteroids (CS), pentoxifylline (PTX) or their combination in
patients with severe alcoholic hepatitis (Maddrey discriminant
function (DF) ≥32) Aims: compare 28-day survival according
to treatment and analyze therapeutic response (Lille model)
Methods: We obtained individual data from 1974 patients,
originating from 9 RCTs, and performed 3 different
meta-anal-yses: CS vs placebo or control treatment (CTL) (n=6 trials),
CS vs PTX (n=2) and CS+PTX vs CS+placebo (n=3) Results:
First analysis: A total of 495 (CS) and 469 (CTL) patients were
included We found no heterogeneity between the trials (test
of interaction not significant: p=0 2) After stratification on
the trial, the HR of mortality at 28 days was 0 643 (95%CI:
0 482-0 855, p=0 002) in CS-treated patients as compared
to CTL After adjustment on DF, creatinine, age and
albu-min, the HR remained significant (HR=0 504, 95%CI 0
.367-0 693, p<.367-0 367-0.367-0.367-01) At day 7, there were more responders
(Lille model<0 45) in CS-treated patients as compared to
CTL (67 4 vs 54 1%, p<0 0001) and the OR of response in
CS-treated patients after adjustment on the trial as compared
to CTL was 1 749 (1 279-2 392, p=0 0005) On available
data at day 28, CS-treated patients had a higher
improve-ment in bilirubin from baseline than CTL: -9 7±0 6 vs -6 9±0 7
mg/dl, p=0 002 Second analysis: 335 (PTX) and 333 (CS) patients were included, without heterogeneity between the tri-als (p=0 3) HR of death at 28 days was 0 643 (0 439-0 943, p=0 002) in CS patients as compared to PTX and was still sig-nificant after adjustment on the same 4 variables (see above):
HR 0 655, p=0 04 There were more responders in CS patients
as compared to PTX: 67 3 vs 48 3%, p<0 0001 and OR for response in CS patients as compared to PTX was 2 29 (1 57-
3 35, p<0 0001) Third analysis: 441 (CS+PTX) and 445 (CS) patients were included, without heterogeneity between trials (p=0 96) After stratification on the trial, the HR of mortality at
28 days was not significant for the combination of CS+PTX as compared to CS alone: HR 0 9, 95%CI 0 63-1 29, p=0 58 The proportion of responders was the same in the two groups:
60 4 vs 64 8% respectively, p=0 25 Thus, OR for response
in CS+PTX patients was not significant (0 835, 95%CI 0
.61-1 61-15, p=0 3) Conclusion: Corticosteroids improve 28-day
sur-vival in patients with severe alcoholic hepatitis with a higher response rate as compared to PTX and CTL This treatment benefit is sustained until the end of therapeutic period The combination of CS and PTX does not add any additional effect Disclosures:
Mark R Thursz - Advisory Committees or Review Panels: Gilead, BMS, Abbott Laboratories, CN-Bio, Altimmune
Timothy R Morgan - Grant/Research Support: Merck, Abbvie, Genentech, ead, Bristol Myers Squibb
Gil-Philippe Mathurin - Board Membership: MSD, Janssen-Cilag, BMS, Gilead, Abvie, Verlyx; Consulting: Roche, Bayer
The following people have nothing to disclose: Alexandre Louvet, Dong Joon Kim, Julien Labreuche, Stephen R Atkinson, Sandeep S Sidhu, John G O’Grady, Robert L Carithers, Marie-Jose Ramond, Charles L Mendenhall, Willis C Mad- drey, Alain Duhamel
1204
Assessment of nutritional status through bioelectrical impedance is a key determinant of outcome in alcoholic hepatitis: a prospective study
Laurent Spahr 1 , Laurence Genton 2 , Laura Rubbia-Brandt 3 , sandra Oropesa 1 , Emiliano G Giostra 1 ; 1 Gastroenterology, University Hospital of Geneva, Geneva, Switzerland; 2 Nutrition, University Hospitals of Geneva, Geneva, Switzerland; 3 Clinical pathology, University Hospitals of Geneva, Geneva, Switzerland
Cas-Alcoholic hepatitis (AH) is a severe complication of alcoholic liver disease with an elevated mortality rate on the short term
in spite of corticosteroids A concomitant poor nutritional status may impact prognosis Aim : We explored the specific role of malnutrition in a prospective cohort of patients with regards to clinical outcome at 6 months Methods : a group of 45 patients admitted to hospital with AH (age 52 9 yrs ; MELD score 19 9 + 5 3 ; Child-Pugh 10 4 + 1 3 ; BMI 26 1 kg/m2 ; C-reactive protein (CRP) 18 8 + 14 9 mg/l ; 62% treated with steroids) were evaluated at baseline using arm muscle circumference (AMC, anthropometric measurement) and phase angle value
by bioelectrical impedance (BIA) This analysis is based on the conductivity properties of body tissues, is performed at bedside, and is a sensitive and non invasive mean to determine nutritional status Clinical condition and alcohol relapse were carefuly monitored Results : Prevalence of malnutrition was 60% (by AMC, < 10th percentile) and 74% (by BIA), statisti-cally higher (p<0 05) as compared to a group of excessive drinkers without AH (n=20) During follow-up, the mortality was 23% Alcohol relapse occurred in 28% of patients Multivari-ate Cox regression analysis showed that phase angle by BIA was an independent predictor of poor outcome at 6 months (p=0 05, 95%CI [0 26-0 99] Other variables such as AMC (p=0 07), age (p=0 83), BMI (p=0 34), MELD score (p=0 35),
Trang 7♦ Denotes AASLD Presidential Poster of Distinction
baseline CRP (p=0 48), and alcohol relapse (p=0 42) were
not related to survival Conclusion : in patients with AH,
mal-nutrition assessed through bioimpedance phase angle is highly
prevalent and has a negative impact on 6-month survival Thus,
a detailed evaluation of malnutrition should be part of alcoholic
liver disease work-up and aggressive nutritional support be
provided in affected patients
Disclosures:
The following people have nothing to disclose: Laurent Spahr, Laurence Genton,
Laura Rubbia-Brandt, Cassandra Oropesa, Emiliano G Giostra
1205
Alcohol Misuse is Associated with Intestinal Mucosal
Dysbiosis and Stool Metabolomic Alterations In Human
Cirrhosis
Jasmohan S Bajaj 1 , Naga Betrapally 2 , Nita Salzman 3 , Douglas
M Heuman 1 , Melanie White 1 , Andrew Fagan 1 , Edith A Gavis 1 ,
Masoumeh Sikaroodi 2 , Oliver Fiehn 4 , Patrick M Gillevet 2 ; 1 VCU
and McGuire VAMC, Richmond, VA; 2 George Mason University,
Manassas, VA; 3 Medical College of Wisconsin, Milwaukee, WI;
4 University of California, Davis, CA
An altered intestinal barrier is associated cirrhosis, especially
alcoholic cirrhosis Intestinal mucosal microbiota structural and
functional alterations in human alcoholic cirrhosis are unclear
Aim: Define the effect of ongoing alcohol misuse on intestinal
mucosal microbiota composition and function, and
antimicro-bial peptides (AMP) in cirrhosis Methods: Compensated
cir-rhotic outpts [active drinkers, Alc & abstinent, NAlc] & healthy
controls (HC) underwent serum collection for endotoxin &
prepped colonoscopy with biopsies from ileum & colon
(ascend-ing, descend(ascend-ing, sigmoid) Stool was collected for microbiota
& metabolomics using GC/MS before the prep Microbiota
was analyzed with multi-tagged sequencing, specifically for
autochthonous bacteria (Lachnospiraceae, Ruminococcaceae,
Clostridiales XIV) Ileal AMPs studied were lysozyme, secretory
phosphlipaseA, Human defensins 4/5, Regenerating
islet-de-rived protein 3 α(HIP/PAP & PSP) & β defensin All values were
compared between groups Results: 56 age-matched subjects
(15 Alc, 13 controls & 28 Nalc) were included Alc and NAlc
pts had similar MELD scores (12 vs 13) All Alc pts had an
AUDIT score >10 and last drink was day prior to sampling
Alc pts had the highest endotoxemia (2 4 vs 1 3 NAlc 0 5
HC EU/ml, p=0 02) On UNIFRAC, microbiota of Alc pts was
significantly different (p<=1 0e-02-0 01) compared to NAlc &
HC in stool, TI, ascending, descending & sigmoid colon HC
& Nalc were different in TI, sigmoid & stool only
Autochtho-nous taxa% was significantly lower in Alc(Table) None of the
AMPs were different between groups (p>0 05) On GC/MS
metabolites focused on bio-energetics (citrate, malate), amino
acids (threonine, ornithine, serine) & pyrimidine intermediates
(ribosine, orotate, hexonate) were significantly lower in Alc
compared to NAlc (0 08-0 48 times lower, p<0 001 for all) &
more than HC (0 04-0 59 times lower, p=6 43E-07-0 01 for
all ) Conclusions: Actively drinking cirrhotis have higher
endo-toxemia likely related to ileo-colonic mucosal dysbiosis This is
accompanied by significant derangements in stool
metabolom-ics towards greater energy and protein consumption likely by
the dysbiotic microbiota compared to abstinent cirrhotics and
healthy controls
*Significantly lower in active drinkers compared to abstinent and Controls #, Significantly lower in cirrhosis compared to controlsDisclosures:
Jasmohan S Bajaj - Consulting: Valeant, Norgine, Intercept, and Abbott; Grant/ Research Support: His institution has received grant funding from Salix and Grifols
Douglas M Heuman - Consulting: Bayer, Grifols, Genzyme; Grant/Research Support: Exilixis, Novartis, Bayer, Bristol Myers Squibb, Scynexis, Ocera, Mann- kind, Salix, Globeimmune, Roche, SciClone, Wyeth, Otsuka, Ikaria, UCB, Cel- gene, Centocor, Millenium, Osiris, AbbVie, Gilead; Speaking and Teaching: Otsuka, Astellas; Stock Shareholder: General Electric
Patrick M Gillevet - Stock Shareholder: BioSpherex LLC, Metabiomics Corp The following people have nothing to disclose: Naga Betrapally, Nita Salzman, Melanie White, Andrew Fagan, Edith A Gavis, Masoumeh Sikaroodi, Oliver Fiehn
1206
Hepatic Overexpression of Splicing Regulator Slu7 liorates Alcoholic Steatosis in Mice via Modification of SIRT1 Alternative Splicing
Ame-Jiayou Wang 1 , Chunki Kim 1 , James P Hardwick 2 , Hong Shen 1,3 , Alvin Jogasuria 1 , Yoonhee Han 1 , Jiashin Wu 1 , Min You 1 ; 1 Pharma- ceutical Sciences, Northeast Ohio Medical University, Rootstown, OH; 2 Integrative Medical Sciences, Northeast Ohio Medical Uni- versity, Rootstown, OH; 3 Liver Diseases, Guangdong Hospital of Traditional Chinese Medicine at Zhuhai, Zhuhai, China
Aberrant hepatic pre-mRNA alternative splicing plays a icant role in liver diseases, but little is known about its involve-ment in development and progression of alcoholic fatty liver disease (AFLD) We have recently identified a new target of ethanol action in liver, namely, pre-mRNA splicing regulator Slu7 In the present study, using adenovirus (Ad)-mediated alteration of hepatic Slu7 expression, we investigated func-tional and causal role of Slu7 in the development of AFLD and explored underlying mechanisms We examined the effects
signif-of Ad-Slu7 and Ad-Slu7shRNA in a mouse model signif-of AFLD created with the Gao-binge (Chronic+binge) ethanol feeding protocol As expected, the ethanol-fed mice robustly elevated levels of hepatic triglyceride and cholesterol in comparison with the pair-fed mice injected with Ad-GFP or Ad-siRNAcon-trol Remarkably, overexpression of Slu7 by Ad-Slu7 injection largely blocked the elevations of triglyceride and cholesterol in the livers of ethanol-fed mice Conversely, knocking down Slu7
by Ad-Slu7shRNA injection did not negate the ethanol effects in mouse liver Mechanically, overexpression of Slu7 mRNA with Ad-Slu7 injection markedly increased the expression of sirtuin
1 (SIRT1) full-length gene and repressed the SIRT1 mRNA native splicing in ethanol-fed mice Conversely, reduction of Slu7 with Ad-Slu7shRNA injection exacerbated the ethanol-me-diated inhibition on full-length SIRT1 gene expression, and stimulated the SIRT1 alternative splicing in respond to ethanol challenge Altogether, our findings suggest that overexpression
alter-of Slu7 ameliorates the ethanol-induced steatosis via tion of SIRT1 mRNA alternative splicing in mice Hence, modu-lation of hepatic Slu7-SIRT1 splicing axis may be beneficial for the prevention and treatment of human AFLD
modifica-Disclosures:
The following people have nothing to disclose: Jiayou Wang, Chunki Kim, James
P Hardwick, Hong Shen, Alvin Jogasuria, Yoonhee Han, Jiashin Wu, Min You
Trang 8Cell Death Biomarkers in Patients with Alcoholic Liver
Disease
Benjamin Woolbright 1 , Winston Dunn 1 , Jody C Olson 1 , Ernst
Malle 2 , Hartmut Jaeschke 1 ; 1 Kansas University Medical Center,
Kansas City, KS; 2 Institute of Molecular Biology, Medical University
Of Graz, Graz, Austria
Chronic alcohol consumption results in progressive destruction
of the liver leading to alcoholic cirrhosis (AC) Alcoholic
hep-atitis (AH) is an acute syndrome that occurs within the greater
spectrum of alcoholic liver disease, with high mortality rates
and limited therapeutic options Neutrophils (PMN) are thought
to contribute to liver injury in both the murine model and in
human patients; however, their role in human patients may be
more complex as the presence of PMN in AH livers is linked
to patient survival We initiated this study to define potential
mechanisms of PMN mediated hepatocyte cell death in AC
and AH patients with the hypothesis that PMN mediated ROS
production leads to increased hepatocellular apoptosis in
human patients Healthy controls (n=10), abstinent patients
with clinically confirmed AC (n=26), and patients with a
clin-ical diagnosis of AH with a recent history of drinking (n=23)
were admitted to the study Blood and plasma were acquired
for analysis of PMN activity, keratin-18 levels (K18), caspase-3
activity, glutamate dehydrogenase (GDH) activity and
cyto-kine levels PMN recruitment was also measured in a
sepa-rate cohort of biochemically matched liver biopsies We first
assessed cell death by measuring necrosis by K18 and
apop-tosis by cleaved K18 (cK18), with the M65 or M30 ELISA
respectively, in plasma of these patients M65 and M30 levels
were elevated in AC and AH patients above controls M65
levels were significantly elevated in AH patients above AC
patients, and these levels were further elevated in non-surviving
patients This occurred despite a negligible increase in plasma
caspase-3 activity or GDH release In spite of this, M30
lev-els were highly diagnostic of AH in the greater population of
overall cirrhotics (AUC by ROC analysis = 0 87, p<0 05), and
M65 levels were prognostic of survival in the AH population
(AUC by ROC analysis = 0 8, p<0 05) We next attempted to
determine if these increases could be linked to PMN activity
CD11b levels and baseline ROS production were measured
by flow cytometry and were elevated in AH neutrophils This
was corroborated by significant increases in circulating IL-8
in the AH population Surprisingly though, PMN recruitment
was similar between AC and AH patients Deposition of ROS
measured by hypochlorous acid modified protein or
chlorotyro-sine staining indicated no difference between the AC and AH
populations As such, it appears that while PMN are active in
blood, there is limited evidence for PMN mediated cell death
in liver In summary, PMN mediated ROS may not be a major
mediator of hepatocyte cell death, although measurements of
K18 may be clinically useful in AH patients
Disclosures:
The following people have nothing to disclose: Benjamin Woolbright, Winston
Dunn, Jody C Olson, Ernst Malle, Hartmut Jaeschke
1208
Prediction And Impact Of Alcohol Abstinence After An Episode Of Alcoholic Hepatitis: A Long-Term Follow-Up Study
Jose Altamirano 1 , Hugo Lopez-Pelayo 2 , Javier Michelena 1 , Lluisa Ortega 2 , Pere Gines 3 , Juan Caballeria 3 , Antoni Gual 2 , Ramon Bataller 4 , Anna Lligoña 2 ; 1 Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; 2 Grup Recerca Addiccions Clinic (GRAC-GRE) Psychiatry Department, Neurosci- ences Institute, Hospital Clínic, Red de Trastornos Adictivos (RTA), Barcelona, Spain; 3 Liver Unit, Hospital Clinic, Institut d’Investiga- cions Biomèdiques August Pi i Sunyer (IDIBAPS), Institut d’Inves- tigacions Biomèdiques August Pi i Sunyer (IDIBAPS), CIBER de Enfermedades Hepáticas y Digestivas (CIBERehd), Barcelona, Spain; 4 Division of Gastroenterology and Hepatology Depart- ments of Medicine and Nutrition University of North Carolina at Chapel Hill, Chapel Hill, NC
Background & Aims: Alcoholic hepatitis (AH) is the most severe form of alcoholic liver disease Most studies are focused on short-term prognosis, while factors associated with long-term survival after an episode of AH are largely unknown We hypothesize that alcohol consumption after an episode of AH heavily impacts patient’s outcome The aims of our study were: 1) to determine impact of complete abstinence from alcohol
on long-term survival and 2) to identify prognostic factors at admission capable of predicting complete abstinence during
long-term follow-up in patients with AH Patients & Methods:
142 patients with biopsy-proven AH admitted at the tal Clinic of Barcelona from 1999 to 2012 were included Demographic, psychiatric and biochemical variables at admis-sion and drinking status during follow-up of all patients were obtained Abstinence was evaluated on the basis of patient’s self-reporting, family member interviews, and determination of the alcohol content in the urine in those attending the hospi-tal addictions unit For the purpose of this study any amount
Hospi-of regular alcohol use after index hospitalization for AH was considered recidivism Univariate and multivariate (Cox and logistic regression) analyses and classification and regression
trees (CART) analysis were used for statistical analysis Results:
Overall mortality was 38% with an median follow-up of 55 months During follow-up complete abstinence was reported in 39% and was associated with long-term survival (p=0 03) On multivariate analysis and after adjustment for baseline prog-nostic scoring systems (MELD and ABIC scores) complete absti-nence was independently associated with survival (p<0 05) When analyzing time frames we did not found significant dif-ferences on survival between abstinent and recidivist patients
at 3, 6 and 12 months of follow-up (p>0 05, for all cases) However, differences in survival became significant after 18 months of follow-up (p=0 045) Age and prior alcoholism treat-ments were independently associated with complete abstinence (p<0 001 and p=0 02, respectively) during follow-up CART analysis generated a simple and practical algorithm based-on the combination of prior alcoholism treatments and age that stratify 2 subgroups of patients with high (65%) and low (26-29%) rates of complete abstinence after an episode of AH
Conclusions: Complete abstinence after an episode of AH itively impacts long-term survival The combination of 2 vari-ables easily assessed at admission might be clinically useful to predict long-term abstinence after an episode of AH Strategies aimed at promoting alcohol abstinence in these patients are mandatory
pos-Disclosures:
Hugo Lopez-Pelayo - Grant/Research Support: Lundbeck, Otsuka Pere Gines - Advisory Committees or Review Panels: Ferring, Ikaria, Promethera, Novartis, Salix; Grant/Research Support: Sequana Medical, Grifols
Trang 9♦ Denotes AASLD Presidential Poster of Distinction
Ramon Bataller - Advisory Committees or Review Panels: Valyx; Speaking and
Teaching: Echosens
The following people have nothing to disclose: Jose Altamirano, Javier
Michel-ena, Lluisa Ortega, Juan Caballeria, Antoni Gual, Anna Lligoña
1209
Outcomes following index hospitalization with
decom-pensated alcohol-related cirrhosis versus severe
alco-holic hepatitis; a retrospective cohort study.
Dev Katarey 1,2 , Jonathan R Potts 1,2 , John D Chetwood 2 , Sumita
Verma 1,2 ; 1 Brighton and Sussex Medical School, Brighton, United
Kingdom; 2 Gastroenterology and Hepatology, Brighton and
Sus-sex University Hospitals Trust, Brighton, United Kingdom
Background: We previously described outcomes in severe
alcoholic hepatitis (SAH), identifying abstinence after
hospi-tal discharge as the only independent predictor of long-term
survival The outcome of patients presenting with other forms
of decompensated alcohol-related liver disease (ALD) is
rela-tively unknown Aim: To assess predictors of inpatient and
long-term survival in patients admitted with decompensated alcohol
related cirrhosis (dARC) as the first presentation of ALD
com-pared to SAH Methods: A retrospective case review was
per-formed for all index hospitalizations with ALD (Apr 2005-Dec
2011) Patients with dARC (Child-Pugh score ≥7 and bilirubin
<80mmol/L) were compared to patients with SAH
(discrimi-nant function ≥32 or biopsy proven) Predictors of inpatient
and long-term survival were determined using binary logistic
regression and Kaplan-Meier method, respectively Results:
177 patients were studied with dARC (n=73, 41 2%) and SAH
(n=104, 58 8%) Those with dARC were older (median age
57-yrs vs 49, P<0 001) There were no significant differences
in admission rates of ascites (overall 80 8%), hepatic
encepha-lopathy (HE; 14 7%), acute kidney injury (AKI; 5 1%), infection
(22 6%) and variceal hemorrhage (6 8%) Inpatient mortality
was similar in both groups (19 2% vs 22 1%, P=0 709) but
length of stay shorter in dARC (6 days vs 11 5, P=0 001) In
dARC the independent determinants of inpatient mortality were
hospital-acquired infection (HAI; OR 20 2 [95% CI 3 5-118],
P=0 001) and HE during hospitalisation (OR 9 3 [1 8-47],
P=0 007) Overall median survival was similar in both groups
(26 months in dARC vs 23 in SAH, P=0 872) with similar
abstinence rates at last follow-up (41 1% vs 34 6%, P=0 431)
In SAH abstinence at last follow-up was the only predictor of
survival (3-yr survival 75 2% in abstainers vs 33 2% in
con-tinued drinkers, P=0 012) However, in dARC abstinence did
not predict long-term survival (median survival 36 months in
abstainers vs 38 in continued drinkers, P=0 949) [Fig 1]
Con-clusions: Inpatient mortality in dALD is predicted by HAI and
HE but not AKI Long-term survival in dARC is not significantly
improved with abstinence in contrast to SAH which suggests a
reduced disease reversibility thereby mandating earlier liver
transplant assessment
Fig 1: Kaplan-Meier survival analysis for dARC and SAH stratified
by abstinence (log-rank P<0 001)
Disclosures:
Sumita Verma - Grant/Research Support: Gilead, Brighton and Hove
Commis-sioners, Dunhill Medical Trust, National Institute of Health Research, Janssen,
Introduction Ceramides are bioactive sphingolipids and are increased in the livers of mice and humans with alcoholic liver disease We previously demonstrated that alcohol-fed mice lacking the major hepatic lipid droplet (LD) protein, Perilipin 2
(PLIN2), are protected from steatosis and have reduced total
hepatic ceramide levels It is not known if ceramides are ent in the LD fraction itself or if alcohol regulates LD ceramide
pres-content Therefore, we investigated the in vivo effect of chronic
alcohol on LD ceramide content in the presence and absence
of PLIN2 Methods Wild-type (WT) and PLIN2 KO mice were
fed a Lieber-DeCarli control (CTRL) or ethanol (ETOH) diet for
4 weeks Livers were dissected for histology, LD isolation, and ceramide analysis Homogenized liver lysate was centrifuged Post-nuclear supernatant was loaded into a sucrose step gradi-ent and ultra-centrifuged The LD fraction was collected, con-firmed by PLIN2 or Perilipin 3 immunoblotting, and sent for mass spectrometric analysis Tissue-specific ceramide synthases (CerS) protein expression was quantified by immunoblotting in CTRL and ETOH WT mice and VL17-A cells incubated in con-
trol or 100mM ethanol media for 48hours Statistical analysis:
t-test, or analysis of variance with post-hoc Newman Keuls
mul-tiple comparison test P≤0 05 is considered significant Results
We again demonstrated that both CTRL and ETOH PLIN2 KO mice are protected from hepatic steatosis Long-chain cera-mides were present in the LD fractions of mice fed both diets Alcohol significantly increased several long-chain ceramide species in LDs of WT mice: C14 (p=0 01), C20 (p=0 001), C20:1 (p=0 002), C22 (p=0 003), C22:1 (p=0 001), C24 (p=0 03), C24:1 (p=0 03), and C26:1 (p=0 045) Addition-ally, there was a non-statistically significant increase in C16, C18, and C18:1 in WT ETOH mice compared with CTRL In the absence of PLIN2, alcohol only increased C22:1 (p=0 02) CerS2 protein expression is similar between control and etha-nol cells while CerS5 protein is reduced in response to ethanol incubation (p=0 03) In mice, CerS2 and CerS5 protein levels are similar in ETOH and CTRL mice In both cells and mice, CerS6 protein is upregulated by 50% in response to ethanol
(p≤0 01) Conclusion Our data demonstrate that ceramides
are present in the LD fraction and that long-chain ceramides are specifically upregulated in the LD fraction in response to alcohol by a PLIN2 dependent mechanism Ceramide synthase
6 may play a key role in alcohol-mediated hepatic ceramide accumulation Future studies will investigate the mechanistic relationship between PLIN2 and ceramide synthetic enzymes
in alcoholic steatosis Disclosures:
The following people have nothing to disclose: Amanke Oranu, Annie Lin, Bianca Williams, Jason Correnti, James Beck, Rotonya M Carr
Trang 10Differential regulation of human
UDP-glucuronosyltrans-ferases (UGTs) during chronic ethanol exposure in a
humanized transgenic UGT1A mouse model
Steffen Landerer, Sandra Kalthoff, Anja Winkler, Christian P
Stras-sburg; University Hospital Bonn, Bonn, Germany
Aims: The inappropriate intake of alcohol leads to alcoholic
liver disease (ALD) usually characterized by the development
of fatty liver, alcoholic hepatitis and fibrosis/cirrhosis, and as
a consequence an increased risk of hepatocarcinogenesis
Eth-anol metabolites and adducts, among them acetic aldehyde,
generated by alcohol dehydrogenase (ADH) and cytochrome
P450 2E1 (CYP2E1)-mediated production of 1-hydroxyethyl
radicals, lead to hypoxic liver injury and the formation of
highly reactive oxygen species (ROS), which are strong
induc-ers of processes driving fibrogenesis Aim of this study was
to examine the regulation of UDP-glucuronosyltransferases 1A
(UGT1As) capable of detoxifying a variety of reactive
metab-olites during chronic ethanol exposure in order to elucidate
the role of high frequency UGT1A polymorphisms (SNPs) in
early stages of ALD Methods: Humanized transgenic (htg)
UGT1A-wild type (WT) and a htgUGT1A-SNP mice,
(contain-ing a haplotype of 10 common UGT1A SNPs) were exposed
to 5% ethanol for 7 months Hepatic RNA was isolated and
UGT1A mRNA expression quantified by TaqMan-PCR Results:
In male htgUGT1A-WT mice, ethanol treatment led to a
sig-nificant induction of hepatic UGT1A1 (12 fold), UGT1A3
(6 fold), UGT1A4 (7 fold), UGT1A6 (9 fold), and UGT1A9
mRNA (7 fold) In contrast, ethanol induced upregulation of
hepatic UGT1A1 and UGT1A9 mRNA was reduced or absent
in htgUGT1A-SNP mice Although UGT1A3, UGT1A4 and
UGT1A6 mRNA expression was highly upregulated (9-43 fold)
in htgUGT1A-SNP mice, absolute expression levels remained
below those of htgUGT1A-WT mice Conclusions: Our study
provides first evidence of human UGT1A gene regulation by
chronic ethanol exposure Characterized by a considerable
activation of UGT1A gene expression in WT mice in contrast to
a differential regulation observed in the presence of SNPs In
htgUGT1A SNP mice absolute expression levels were
consider-ably decreased In conclusion, a common UGT1A haplotype,
observed in 9% of the white population, significantly impairs
the physiological responsiveness towards the oxidative stress
inducer ethanol and therefore represents a potential risk factor
for the development of hepatic fibrosis in ALD
Disclosures:
Christian P Strassburg - Advisory Committees or Review Panels: Novartis, Roche;
Speaking and Teaching: Novartis, Merz, MSD, Falk Pharma, BMS, Abbvie
The following people have nothing to disclose: Steffen Landerer, Sandra Kalthoff,
Anja Winkler
1212
Alcoholic Hepatitis and Disease Severity Are Associated
With Distinct Shifts in Fecal Microbial Ecology
Puneet Puri 1 , Kalyani Daita 1 , Faridoddin Mirshahi 1 , Suthat
Liang-punsakul 2 , Naga P Chalasani 2 , David W Crabb 2 , Vijay Shah 3 ,
Patrick S Kamath 3 , Gregory J Gores 3 , Barry P Katz 2 , Masoumeh
Sikaroodi 4 , Svetlana Radaeva 5 , Patrick M Gillevet 4 , Arun J
Sanyal 1 ; 1 Virginia Commonwealth University, Richmond, VA;
2 Indiana University School of Medicine, Indianapolis, IN; 3 Mayo
Clinic, Rochester, MN; 4 George Mason University, Manassas, VA;
5 National Institute on Alcoholism and Alcohol Abuse, Bethesda,
MD
BACKGROUND: Intestinal dysbiosis is linked to alcoholic
hep-atitis (AH) However, changes in fecal microbiome in heavy
drinking alcoholics without liver disease and with AH are
unknown AIMS: (1) To characterize microbial ecology in
sub-jects with AH and compare it to heavy drinking controls (HDC), and (2) To relate changes in microbial ecology with disease
severity METHODS: Subjects with heavy drinking (alcohol >60
g/d in men and >40 g/d in women, for >6 months) and no evidence of liver disease (HDC), or bilirubin > 2mg/dL and AST>50 (AH) were studied MELD score defined severity of AH: moderate (≤ 20, MAH) or severe (>20, SAH) Fecal 16s rRNA sequencing was performed UNIFRAC principal compo-nent, pairwise, multiple group comparisons, and regression analyses were performed The impact of microbial shifts was determined by linear discriminant analysis (LDA) Effect Size
(LEfSe) RESULTS: Of 54 enrolled subjects (59% males, mean
age 46±12 years), 20 were HDC and 34 AH (10 MAH, 24 SAH) Bacteroidetes and Firmicutes were most abundant phyla
in both HDC and AH Fecal microbiome changes: AH had high abundance of Fusobacteria (p=0 004), Firmicutes Nega-tivicutes and Selenomonadales (both p=0 01), while Firmicutes Clostridia and Clostridiales were diminished (both p=0 0001) Impact of microbial shifts: The LDA scores >3 indicate large effect size of microbial shifts Accordingly, HDC was enriched with members of Firmicutes phylum (Clostridia, Clostridiales, Lachnospiraceae, and Ruminococcaceae) In contrast, MAH was enriched with Enterococcaceae and SAH with Fusobac-teriales, Enterobacteriaceae, and Actinomyces Relation with disease severity: Compared to HDC, increased Fusobacte-ria (p=0 001) and Firmicutes Negativicutes (p=0 01), and decreased Firmicutes Clostridiales (p=0 0003) were exclusive
to SAH Fimicutes Baclilli correlated with Discriminant Function (DF) (r2=0 43, p=0 0004) and MELD score (r2=0 07, p=0 04)
In contrast, Firmicutes Clostridia was inversely related to MELD score (r2=0 33, p<0 0001), Child score (r2=0 22, p=0 0007) and DF (r2=0 22, p=0 02) The strong relationship in HDC of Bacteroidetes and Actinobacteria (ρ= -0 84), and Actinobac-teria and Firmicutes (ρ=0 73) was not observed in MAH and SAH The inverse relation of Proteobacteria and Firmicutes (ρ=
-0 62, p=0 001) characterized SAH CONCLUSIONS: (1) AH
is associated with enrichment of Fusobacteria (2) Clostridia may be protective against the development of severe AH (3) Perturbations in microbial ecology are associated with severe
AH These data offer promise for disease biomarkers with implications for potential therapeutics
Trang 11♦ Denotes AASLD Presidential Poster of Distinction
1213
The Role of Protein Carbonylation in Progression of
Alcoholic Liver Disease in Male GSTA4-4 -/- Mice
Colin T Shearn 1 , Martin J Ronis 2 , Kelly E Mercer 3 , Casey
Pul-liam 2 , Dennis R Petersen 1 ; 1 Department of Pharmaceutical
Sci-ences, University of Colorado Anschutz Medical Campus, Aurora,
CO; 2 Department of Pharmacology & Therapeutics, Louisiana State
University Health Sciences Center, New Orleans, CO; 3 University
of Arkansas for Medical Sciences, Little Rock, AR
Alcoholic liver injury (ALD) is considered to result from a
multi-ple hit mechanism in which the first hit, reversible development
of fatty liver, is followed in susceptible individuals by several
other hits, thought to include various forms of oxidative stress
and endotoxemia, ultimately resulting in progression of
pathol-ogy to inflammation, fibrosis and hepatocellular carcinoma In
the current study, we examined the potential role of reactive
short chain aldehyde (SCA) products of lipid peroxidation in
progression of ALD by examining the development of ALD in
male wild type SV/J mice (WT) and SV/J mice lacking the
enzyme glutathione-S-transferase (GST)A4-4 (GST -/-) which is
responsible for detoxification of SCAs such as
4-hydroxynon-enal (4-HNE) Mice, 13 wks old, were fed chow (N = 10-12)
or high fat Lieber DeCarli liquid diets containing 28% calories
as ethanol (EtOH)(N = 18-20) for 120 d At the end of the
study, half of each group were also acutely challenged with
saline or an EtOH binge (3 g/kg given at a concentration of
30% v/v, i g ) and all mice were sacrificed 12 h later Both
WT and GST-/- EtOH-treated mice developed steatosis and
had elevated serum ALT values In the EtOH-treated groups,
expression of inflammatory markers TNFa and IFNg mRNA
and T cell markers CD4 and FOXP3 were elevated in the
GST-/- mice compared to WT (P<0 05) In addition, the
prolifera-tion marker Ki67, matrix remodeling genes MMP9, MMP13
and the fibrosis marker collagen 1A mRNA and collagen
stain-ing were all higher in EtOH GST-/- compared to EtOH WT
mice (P<0 05) mRNA encoding the B cell marker B220 and
mRNA for CD138 a marker of fully differentiated antibody
pro-ducing plasma cells were elevated by EtOH treatment (P<0 05)
only in the GST-/- mice Acute binge EtOH increased necrosis
and reduced expression of IL-6 in both groups (P<0 05) but
the effect was greater in EtOH GST-/- than EtOH WT mice
(P<0 05) Overall carbonylation as assessed by
immunohisto-chemistry was increased in both WT and GST-/- mice treated
with EtOH but was more prevalent in the GST-/- mice
Com-prehensive LC-MS/MS analysis of carbonylation revealed a
total of 1022 proteins of which 189 were unique to the
GST-/- group Using a MASCOT cutoff score of 24 and excluding
peptides with C-terminal lysine modifications, the site of
alde-hyde modification was identified on an additional 33 peptides
from both models These data suggest long term adaptation to
EtOH in WT mice not observed in GST-/- mice SCA play a
role in inflammatory responses due to ETOH and that the effects
of EtOH on B cell differentiation and autoimmune responses
may be secondary to formation of carbonyl adducts
Disclosures:
The following people have nothing to disclose: Colin T Shearn, Martin J Ronis,
Kelly E Mercer, Casey Pulliam, Dennis R Petersen
1214
PNPLA3 G/G genotype doubles the mortality risk of patients with portal hypertension due to fatty liver dis- ease
Mattias Mandorfer, Albert Stättermayer, Bernhard Scheiner, Rafael Paternostro, Nastja Matko, Dorothea Plaschka, Constantin Vier- ziger, Laura Zechmeister, Wolfgang Sieghart, Michael H Trauner, Thomas Reiberger, Harald Hofer, Markus Peck-Radosavljevic, Peter Ferenci, Arnulf Ferlitsch; Dept of Internal Medicine III, Div
of Gastroenterology & Hepatology, Medical University of Vienna, Vienna, Austria
Background and aim The rs738409 C>G p I148M variant in
the patatin-like phospholipase domain containing 3 (PNPLA3) gene promotes triglyceride accumulation in hepatocytes and liver fibrosis, with more consistent effects among patients with fatty liver disease when compared to viral hepatitis However, its prognostic impact in patients who have already developed advanced liver disease is less clear Thus, we aimed to inves-tigate the influence of PNPLA3 on transplant-free survival (TFS)
in a thoroughly characterized cohort of patients with advanced liver disease and portal hypertension (hepatic venous pres-sure gradient [HVPG]≥6mmHg) due to viral hepatitis or fatty
liver disease Methods We performed a retrospective analysis
in prospectively characterized patients who underwent HVPG measurement at the Medical University of Vienna through
2013 The effect of PNPLA3 G/G genotype on TFS was tigated using Cox models adjusted for age, HVPG and MELD
inves-Results Three hundred and seventy-two patients had portal hypertension due to viral hepatitis (n=231 [62%]) or fatty liver disease (n=141 [38%]; alcoholic: 104[28%]/non-alcoholic: 37[10%]) The median HVPG and MELD were 16 (interquartile range [IQR]:10) mmHg and 9 32 (IQR:4 97) points, respec-tively Fifty-six (15%) patients had the PNPLA3 G/G genotype, while 163 (44%) and 153 (41%) had the C/C and the C/G genotype, respectively In the overall cohort, we observed a trend toward reduced TFS among patients with the PNPLA3 G/G genotype (hazard ratio [HR]:1 53; 95% confidence
interval [95%CI]:0 95-2 46; P=0 082), after adjusting for age (per year; HR:1 03; 95%CI:1 01-1 05; P=0 001), HVPG (per mmHg; HR:1 06; 95%CI:1 02-1 09; P=0 002) and MELD (per point; HR:1 01; 95%CI:0 95-1 08; P=0 675) While PNPLA3
G/G genotype was not associated with TFS in the subgroup
of patients with viral hepatitis (adjusted HR:0 97; 95%CI:0
.41-2 41-27; P=0 937), we observed a strong association between
PNPLA3 G/G genotype and TFS in patients with fatty liver
disease (adjusted HR:2 04; 95%CI:1 11-3 76; P=0 022)
Interestingly, even in patients with fatty liver disease who had already developed clinically significant portal hypertension (CSPH; HVPG≥10mmHg), PNPLA3 G/G genotype reduced
TFS (adjusted HR:2 09; 95%CI:1 1-4; P=0 024) Conclusions
The PNPLA3 G/G genotype doubles the mortality risk of patients with portal hypertension due to fatty liver disease, even after the development of CSPH Further studies investigating the underlying pathophysiological mechanism that accelerates pro-gression at this advanced stage of the disease are warranted Disclosures:
Mattias Mandorfer - Consulting: Janssen; Speaking and Teaching: AbbVie, ead, Janssen, Boehringer Ingelheim, Bristol-Myers Squibb, Roche
Gil-Bernhard Scheiner - Speaking and Teaching: Gilead Wolfgang Sieghart - Grant/Research Support: Bayer Schering Pharma, Bayer Schering Pharma, Bayer Schering Pharma, Bayer Schering Pharma; Speaking and Teaching: Bayer Schering Pharma, Bayer Schering Pharma, Bayer Schering Pharma, Bayer Schering Pharma
Michael H Trauner - Consulting: Albireo, Falk, Phenex, Gilead, Novartis, MSD; Grant/Research Support: Falk, Albireo, Intercept
Thomas Reiberger - Consulting: Xtuit; Grant/Research Support: Roche, Gilead, MSD, Phenex, Philipps; Speaking and Teaching: Roche, Gilead, MSD
Trang 12Harald Hofer - Advisory Committees or Review Panels: Gilead, Abbvie; Speaking
and Teaching: Janssen, BMS, Gilead, Abbvie
Markus Peck-Radosavljevic - Advisory Committees or Review Panels: Bayer,
Inter-cept, Gilead, Janssen, BMS, AbbVie; Consulting: Bayer, Shionogi,
Boehring-er-Ingelheim, ONO Pharma, Eli Lilly, AbbVie; Grant/Research Support: Bayer,
Gilead, MSD, AbbVie; Speaking and Teaching: Bayer, Gilead, MSD, AbbVie
Peter Ferenci - Advisory Committees or Review Panels: Idenix, Gilead, MSD,
Janssen, Salix, AbbVie, BMS, Wilson Therapeutics; Patent Held/Filed: Madaus
Rottapharm; Speaking and Teaching: Gilead, Roche
The following people have nothing to disclose: Albert Stättermayer, Rafael
Pater-nostro, Nastja Matko, Dorothea Plaschka, Constantin Vierziger, Laura
Zechmeis-ter, Arnulf Ferlitsch
1215
Hepatic Cell Death Caused by Fungi as well as Bacteria
Is Mediated by ROS Induction of Nuclear
Transglutami-nase in Hepatic Cells
Soichi Kojima 1 , Ronak Shrestha 2,1 , Xian-Yang Qin 1 , Susumu
Kajiwara 2 ; 1 Micro-Signaling Regulation Technology Unit, RIKEN,
Wako, Japan; 2 Department of Life Science, Tokyo Institute of
Tech-nology, Yokohama, Japan
Background: The liver acts as the first barrier to the spread of
fungi and bacteria present in intestine However, in ASH and
NASH patients, these fungi and bacteria are known to invade
gastrointestinal mucosa to reach the liver and cause severe
fun-gal and bacterial infections, causing hepatic injury Previously,
we reported an important role of induced nuclear
transgluta-minase (TG)2 activity in ethanol- and free fatty acid-induced
liver injury via crosslinking and silencing Sp1 transcription
factor thus decreasing expression of c-Met essential for
sur-vival of hepatic cells Aim: In this paper, by investigating the
cellular activity of TG2 in human hepatic cell line (HC cells)
following co-incubation with Candida species and E coli, we
explored the hypothesis that these fungi and bacteria might
induce the nuclear activity of TG2 in hepatic cells leading to
hepatic cell death Methods: Nuclear TG2 activity and ROS
were visually monitored using 5-(biotinamido)pentylamine
(5BAPA) and CM-H2DCFDA ROS species were determined
using ESR Results: Co-incubation of HC cells with pathogenic
fungi (C albicans and glabrata) dose- and time-dependently
enhanced nuclear TG2 activity 7-fold within 24 h and caused
hepatic cell death accompanying caspase-3 within 48 h In
TG2-/- primary hepatocytes the opportunistic Candida species
failed to show the similar effect Similar cell death was induced
by E coli BL21, but not by Sacchromyces cerevisiae Because
induction of nuclear TG2 was reproduced in the co-cultures
using a transwell separated by dialysis membrane, but not by
fungal or bacterial conditioned media (CM), we suspected a
small molecular weight substance(s) with a short half life time
and explored the possibility of ROS as a mediator, and found
that the same phenomenon was reproduced by H2O2, while
N-acetyl cysteine (NAC), an inhibitor of ROS, blocked it
Induc-tion of nuclear TG2 was accompanied by emergence of ROS
in the same nucleus and OH radical was detected in freshly
harvested CM by ESR The NADPH oxidase gene mutant of C
glabrata (named Cgnox1) failed to induce similar phenomena
Conclusion: This paper shows, for the first time, an association
of ROS-producing fungi like C albicans and glabrata as well
as E Coli with enhanced nuclear TG2 activity in hepatic cells
leading to apoptosis, illustrating the impact of ROS-generating
pathogens in inducing and/or exacerbating nuclear
TG2-re-lated liver injuries, which may explain a molecular mechanism
of hepatic injury seen in ASH/NASH patients We are now
underway of animal experiments to strengthen this conclusion
Disclosures:
The following people have nothing to disclose: Soichi Kojima, Ronak Shrestha,
Xian-Yang Qin, Susumu Kajiwara
Purpose: Alcoholic hepatitis (AH) is a clinical entity with high short-term mortality Acute kidney injury (AKI) often complicates the clinical course and negatively impacts outcome in AH pts Data on AKI in AH pts are limited We performed this study in well characterized AH pts to evaluate a) prevalence and pre-dictors of of AKI and b) its impact on short-term (90 d) mortal-
ity Methods: Individual patient data obtained from two centers
on well characterized hospitalized AH pts for demographics, MELD labs, inpatient AKI (defined by AKIN criteria), bacterial infections, length of hospital and ICU stay, and 90 d mortality
Results: On pooled data (n=482), 117 from Center 1 (median age 47 yrs , 66% m, 80% white, 52% cirrhosis, 68% systemic inflammatory response syndrome at admission [SIRS]) and 365 from Center 2 (median age 45 yrs , 98% m, 100% Asians, 85% cirrhosis, 65% SIRS), 163 (34%) patients developed inpatient AKI, with similar prevalence at center 1 (35%) vs 2 (33%) Proportion of AKIN stages I-III were 61, 15, 24% for center 1 and 23, 29, 48% for center 2, respectively (P<0 001) Patients developing AKI had more frequent encephalopathy (34 vs 12%), infection (44 vs 30%) and SIRS (82 vs 57%) with higher median MELD (24 vs 20) and ABIC scores (8 2 vs
7 5), P<0 001 for all Patients with (n=311) compared to out (n=164) SIRS had rate of AKI (42% vs 18%, p<0 001), but similar comparing infected (144) vs non-infected (167) SIRS patients (46 vs 38%, P=0 18) Interestingly, among SIRS criteria, just tachycardia was more common in pts with vs without AKI (72 vs 53%, p<0 0001) On COX regression analysis, AKI was predicted by SIRS [3 13 (1 92-5 10)] and only heart rate SIRS criterion [2 34 (1 52-3 60)] AKI pts vs without AKI had poorer 90 d survival (49% vs 29%, Log Rank P<0 001) with 59, 41, and 31% survival AKIN I-III respec-tively, P=0 31 AKI pts with (n=24) vs without (n=139) dialysis had worse survival (25 vs 55%, P<0 001) AKI pts used more hospital resources with longer median hospital stay (14 vs 9 d), dialysis (15 vs 2 %), and ICU stay (55 vs 22%), P<0 001 for all On cox-regression analysis after controlling for disease severity and center, inpatient AKI increased 90d mortality by about 1 5 fold, other predictors being male gender and cirrho-
with-sis Conclusion: AKI occurs frequently during hospitalization
in AH pts and heavily impacts survival Presence of SIRS at admission, especially tachycardia, is associated with onset of inpatient AKI Patients requiring dialysis have a bad outcome Maneuvers aimed at preventing SIRS associated AKI in AH are urgently needed to improve the outcome of this severe clinical condition
Disclosures:
Ramon Bataller - Advisory Committees or Review Panels: Valyx; Speaking and Teaching: Echosens
The following people have nothing to disclose: Sujan Ravi, Rakhi Maiwall, Shiv
K Sarin, Jose Altamirano, Ashwani Singal
Trang 13♦ Denotes AASLD Presidential Poster of Distinction
1217
Adipose-specific lipin1 overexpression protects mice
against alcohol-induced liver injury
Wenliang Zhang, Wei Zhong, Qian Sun, Xinguo Sun, Zhanxiang
Zhou; Center for transnational biomedical research, University of
North Carolina Greensboro, Kannapolis, NC
Background: Excessive fatty acid release from the white
adi-pose tissue (WAT) contributes to the development of alcoholic
liver disease (ALD) Lipin1 is a co-regulator of DNA-bound
tran-scription factors and a phosphatidic acid (PA) phosphatase
(PAP) enzyme that dephosphorylates PA to form diacylglycerol
(DAG), but it was dramatically reduced by alcohol in the WAT
This study aimed at determining the role of adipose lipin1 in
alcohol-induced lipodystrophy and the development of ALD
Methods: Transgenic mice overexpressing lipin1 in adipose
tissue (aP2-LPIN1 Tg) were produced Wild type (WT) and
aP2-LPIN1 mice were fed a Lieber-DeCarli alcohol or isocaloric
maltose dextrin control liquid diet for 8 weeks to induce ALD
Blood parameters, hepatic steatosis, ER stress, apoptosis, and
lipid metabolism were measured after 8-week alcohol feeding
Results: Alcohol feeding to WT mice resulted in liver damage
as indicated by elevated plasma alanine aminotransferase
(ALT, 237 68 ± 142 20 U/L) and aspartate
aminotransfer-ase (AST, 259 86 ± 142 44 U/L), which was significantly
alleviated by overexpression of lipin1 in adipose tissue (ALT,
44 62 ± 26 05 U/L; AST, 85 85 ± 26 76 U/L) Alcohol
feed-ing significantly reduced epididymal WAT (eWAT) mass in
association with increased lipolysis and inhibited lipogenesis
in the eWAT of the WT mice, which was attenuated in the
aP2-LPIN1 Tg mice Adipose-specific lipin1 overexpression also
attenuated alcohol-reduced plasma leptin levels Alcohol
feed-ing induced hepatic lipid accumulation and down-regulation of
beta-oxidation genes in the liver of WT mice Overexpression
of lipin1 in adipose tissue significantly alleviated
alcohol-in-duced lipid accumulation and abolished the down-regulation of
beta-oxidation genes in the liver Alcohol feeding also resulted
in significant hepatic ER stress and apoptosis in the WT mice,
which was significantly attenuated in the aP2-LPIN1 Tg mice
Conclusions: The results suggest that overexpression of lipin1
in adipose tissue alleviates alcohol-induced liver injury through
restoration of WAT lipid storage function and lipin secretion
Key words: alcoholic liver disease, lipin1, liver injury, adipose
tissue
Disclosures:
The following people have nothing to disclose: Wenliang Zhang, Wei Zhong,
Qian Sun, Xinguo Sun, Zhanxiang Zhou
1218
Alcohol-induced activation of the unfolded protein
response in Paneth cells induces small intestinal
inflam-masome activation, pro-apoptotic signaling and IL-17
production
Benedek Gyongyosi, Patrick P Lowe, Arvin Iracheta-Vellve,
Abhishek Satishchandran, Aditya Ambade, Gyongyi Szabo;
Med-icine, UMass Medical School, Worcester, MA
Purpose: Alcohol-induced intestinal barrier dysfunction and
microbiome changes contribute to the development of alcoholic
liver disease Previous studies found alcohol-induced changes
in tight junction proteins both in the proximal and distal small
bowel However, it is yet to be explored how alcohol affects
the intestine’s own innate defense mechanisms, particularly the
secretory Paneth cells which are highly susceptible to
endoplas-mic reticulum (ER) stress In this study, we hypothesized that
chronic alcohol results in inflammatory changes in the small
intestine Methods: Mice received a Lieber-DeCarli ethanol or isocaloric diet for 10 days followed by an oral gavage of 5g/
kg ethanol or sugar solution, respectively Gut tion and ER stress inhibition was achieved by antibiotics or 4-phenylbutiric acid (4-PBA) administration, respectively Small intestinal unfolded protein response (UPR), pro-apoptotic and pro-inflammatory signaling were assessed by immunoblotting Paneth cell-related antimicrobial gene expressions were mea-sured by quantitative polymerase chain reaction Paneth cell numbers were assessed by periodic acid-Schiff staining and lysozyme immunohistochemistry Interleukin (IL)-17 release was measured in ethanol-stimulated isolated crypts Results: Chronic ethanol feeding resulted in selective induction of the C/EBP homologous protein (CHOP) arm of UPR signaling in the prox-imal but not in the distal small intestines (SI) of alcohol-fed mice, localizing to the Paneth cells of the crypts of Lieberkühn There was no activation in the other arms of UPR (ATF6 cleav-age, Xbp1 splicing) This was accompanied by an increase
decontamina-in Paneth cell numbers, antimicrobial gene expression and IL-17 protein expression in the proximal and not in the distal
SI Binge alcohol exposure after 10 days alcohol feeding ther increased pro-apoptotic signaling with phosphorylation of eukaryotic translation initiation factor 2A (eIF2a) and induction
fur-of CHOP-related downstream targets, GADD34, Bax and Bim Alcohol-induced ER stress also activated the inflammasome, as indicated by caspase-1 and IL-18 cleavage Alcohol-induced inflammasome activation and ER stress induction were inhib-ited by antibiotics or 4-PBA administration In vitro, ethanol induced IL-17 release from isolated crypts that was mimicked
by thapsigargin and prevented by 4-PBA, indicating an ER stress-mediated mechanism Conclusions: Alcohol intake upreg-ulates small intestinal innate defense mechanisms by increasing Paneth cell numbers, antimicrobial gene expression, and IL-17 release We show that binge alcohol-induced small intestinal inflammasome activation occurs via microbiome-induced ER stress
Uni-The following people have nothing to disclose: Benedek Gyongyosi, Patrick P Lowe, Arvin Iracheta-Vellve, Abhishek Satishchandran, Aditya Ambade
1219
Hepatic SIRT1 is Dispensable for Adiponectin-Mediated Amelioration of Bile Acid Metabolism Abnormalities and Liver Dysfunction in Ethanol-Fed Mice
Chunki Kim 1 , Yoonhee Han 1 , Alvin Jogasuria 1 , Jiayou Wang 1 , Xudong Hu 1,3 , Hong Shen 1,2 , Jiashin Wu 1 , Min You 1 ; 1 Pharma- ceutical Sciences, Northeast Ohio Medical University, Rootstown, OH; 2 Liver Diseases, Guangdong Hospital of Traditional Chinese Medicine at Zhuhai, Zhuhai, China; 3 Biology, Shanghai University
of Traditional Chinese Medicine, Shanghai, China
Adiponectin, an adipocyte-derived circulating protein, elicit multiple beneficial effects on the liver injury induced by eth-anol in mice We have previous shown that sirtuin 1 (SIRT1)
is a downstream target of adiponectin signaling in cultured hepatocytes and in mouse liver In the present study, we sought
to determine the requirement of hepatocyte-specific SIRT1 in mediating the adiponectin effects in alcoholic fatty liver injury, utilizing hepatocyte-specific SIRT1 knockout (SIRT1LKO) mice
Trang 14and their littermate control (WT) mice Eight groups of matched
SIRT1LKO and WT mice were fed either an ethanol-containing
or control diet using a chronic-binge ethanol feeding
proto-col Four groups of SIRT1LKO and WT mice fed either with
or without ethanol were injected with full length mammalian
adiponectin (flAcrp, 0 5 μg/g body weight/day) during the
feeding period As expected, ethanol administration to WT
mice led to development of steatosis, inflammation and liver
injury, and alcoholic fatty liver was exaggerated in the
etha-nol-fed SIRT1LKO mice Surprisingly, administration of flAcrp
effectively alleviated alcoholic fatty liver injury as revealed
by ameliorated neutrophilic inflammation, reduced hepatic
triglycerides and cholesterol levels and diminished serum
AST and ALT levels in both ethanol-fed WT and ethanol-fed
SIRT1LKO mice Mechanistically, administration of flAcrp
nor-malized the ethanol administration-induced elevations of both
hepatic pool and blood levels of bile acids in both WT and
SIRT1LKO mice In concordance with the normalized bile acid
metabolism, administration of flAcrp attenuated hepatic
oxida-tive stress, ameliorated hepatic lipin 1 gene expression and its
alternative splicing, which in turn, led to increased fatty acid
oxidation, reduced lipogenesis, limited accumulation of toxic
bile, diminished inflammation, and ameliorated
steatohepati-tis in the livers of both WT and SIRT1LKO mice after ethanol
challenge Taken together, these data demonstrate that
hepato-cyte-specific SIRT1 is dispensable for the adiponectin-mediated
amelioration of bile acid metabolism abnormalities and liver
injury in mice following ethanol administration Our novel
find-ings shed light on the mechanism underlying the protective
action of adiponectin against development and progression of
alcoholic liver disease
Disclosures:
The following people have nothing to disclose: Chunki Kim, Yoonhee Han, Alvin
Jogasuria, Jiayou Wang, Xudong Hu, Hong Shen, Jiashin Wu, Min You
1220
The chemical chaperone 4-phenylbutyric acid prevents
ethanol-induced liver injury in diabetic KK-Ay mice
Maiko Suzuki, Kazuyoshi Kon, Kenichi Ikejima, Akira Uchiyama,
Kumiko Arai, Tomonori Aoyama, Shunhei Yamashina, Sumio
Watanabe; Department of Gastroenterology, Juntendo University
School of Medicine, Tokyo, Japan
Background: Diabetes mellitus is a potential exacerbating
fac-tor in alcoholic liver injury Ethanol (EtOH) exposure promotes
endoplasmic reticulum (ER) stress in several organs; however,
the role of ER stress on EtOH-induced liver injury has not been
fully understood The chemical chaperone 4-phenylbutyric
acid (PBA) reduces ER stress Our Aim in this study was to
investigate the effect of PBA on EtOH-induced liver injury using
diabetic KK-Ay mice Methods: Male KK-Ay mice at 8
weeks-old were fed Lieber-DeCarli diet containing 5% EtOH for 10
days Some mice were given PBA (120 mg/kg BW, i p , daily)
during the feeding period Mice were sacrificed at day 11,
and some mice were sacrificed at 9 h after a single gavage of
EtOH (4 g/kg BW) Control mice were given a single gavage
of isocaloric dextrin after a pair-fed control diet Oxidative
stress and apoptosis were assessed by immunohistological
staining for 4-hydroxy-2-nonenal (4-HNE) and caspase-cleaved
cytokeratin 18, respectively Serum ALT levels were measured
colorimetrically Hepatic expression of mRNA for spliced x-box
binding protein-1 (sXBP-1), heme oxygenase (HO)-1, tumor
necrosis factor (TNF)-α, and interleukin (IL)-6 were quantified
by RT-PCR Results: While chronic EtOH feeding alone did
not elevate serum ALT levels, hepatic sXBP-1 mRNA was
sig-nificantly elevated (2 0±0 4-fold vs controls, p<0 05) After
binge, serum ALT was significantly elevated to 107±19 IU/L compared with controls (23±1 IU/L, p<0 05), and massive fat accumulation with inflammation were observed Significant increase of 4-HNE was observed in 21 4±3 4% of hepatocytes compared with control (0 2±0 0%), and expression of mRNA for HO-1, TNFα and IL-6 was also significantly increased, val-ues reaching 14 0±0 9, 2 7±0 2 and 9 8±1 6-fold vs controls (p<0 05), respectively The percentage of apoptotic cells sig-nificantly increased (3 9±1 6%, p<0 05) Treatment with PBA during chronic EtOH exposure completely inhibited expression
of sXBP-1 mRNA to control levels (1 1±0 1-fold vs controls) PBA significantly prevented elevation of serum ALT after binge (62±2 IU/L, p<0 05), and attenuated fat and inflammation PBA also reduced the percentage of 4-HNE positive cells (12 1±2 5%, p<0 05), and reduced the expression of mRNA for HO-1, TNFα, and IL-6 (8 1±1 9, 1 7±0 3 and 6 0±1 3-fold, p<0 05, respectively) Finally, the treatment with PBA signifi-
cantly decreased apoptotic cells (2 0±0 5%, p<0 05)
Con-clusions: Our findings indicated that chronic-binge EtOH-fed KK-Ay mouse is a suitable model for alcoholic liver injury with diabetes mellitus Prevention of ER stress by chemical chaper-one is a potential therapy for alcoholic liver injury
Disclosures:
The following people have nothing to disclose: Maiko Suzuki, Kazuyoshi Kon, Kenichi Ikejima, Akira Uchiyama, Kumiko Arai, Tomonori Aoyama, Shunhei Yamashina, Sumio Watanabe
Background The liver is a primary target of alcohol toxicity, but chronic alcohol abuse can damage several other organs, including the lung In fact, previous studies have shown that chronic ethanol exposure increases the incidence, severity, and mortality of sepsis-induced acute lung injury (ALI) Furthermore, recent work by this group has indicated that liver injury caused
by alcohol may contribute to lung pathology To date, most animal models investigating the role of alcohol in ALI require administration of an exogenous inflammatory stimulus (e g , LPS); whether or not alcohol exposure is sufficient to induce pulmonary changes is unclear The ‘NIAAA’ model of acute-on-chronic alcohol exposure is a recently developed model that appears to better recapitulate alcoholic (steato)hepatitis The impacts of this liver model on lung pathology have not been explored; the goal of this study was to therefore char-acterize simultaneously the effects of acute-on-chronic alcohol
on the liver and lung Methods 10 W male C57Bl6/J mice
were exposed to alcohol on the NIAAA protocol: 10 days liquid diet, followed by a bolus gavage (5 g/kg) Animals were sacrificed 9 or 24 hours after the gavage, and liver and lung tissue, plasma, and bronchoalveolar lavage fluid (BALF) were collected for analysis Injury was determined biochemi-cally and histologically Inflammatory cytokine and chemokine
production were also quantitated Results As expected,
acute-on-chronic alcohol feeding caused significant steatohepatitis, characterized by increased plasma AST/ALT, as well as fat and neutrophil accumulation Interestingly, acute-on-chronic
Trang 15♦ Denotes AASLD Presidential Poster of Distinction
alcohol exposure also caused transient lung injury Similar to
liver, this injury was characterized by a predominantly
neutro-philic inflammatory response in lung tissue and BALF Indices
of the pulmonary inflammasome were also elevated by
acute-on-chronic alcohol in the lung, analogous to previous findings
in the liver Conclusions Taken together, these data indicate
that the newly developed NIAAA model of alcoholic (steato)
hepatitis also develops concomitant lung damage These results
indicate, for the first time, that alcohol exposure is sufficient
to induce pulmonary inflammatory injury and emphasize the
parallel (and potentially interdependent) damage between the
liver and the lung after alcohol exposure This work was
sup-ported, in part by NIH grants R01AA021978 (GEA, PI) and
R01AA013353 (JR, PI)
Disclosures:
Jesse Roman - Consulting: Boehringher Ingelheim; Grant/Research Support:
Gil-ead, Actelion, Fibrogen, Novartis, Promedior
The following people have nothing to disclose: Lauren G Poole, Juliane I Beier,
Edilson Torres-Gonzalez, Anwar Anwar-Mohamed, Nikole L Warner, Christine
E Dolin, Calvin T Nguyen-Ho, Jeffrey D Ritzenthaler, Gavin E Arteel
1222
Proteome Dynamics reveals that High Density
Lipo-protein Dysfunction in Nonalcoholic Steatohepatitis
is related to Increased Degradation of HDL proteins
involved in Reverse Cholesterol Transport.
Takhar kasumov 2 , Kwangwon Lee 2 , Abdullah Osme 2 , Jaividhya
Dasarathy 3 , Ling Li 4 , Belinda Willard 4 , Iryna Kalinina 4 , Srinivasan
Dasarathy 4 , Jonathan D Smith 1 , Arthur J McCullough 4 ; 1
Gas-troenterology, Cleveland Clinic, Cleveland, OH; 2 Pharmaceutical
Sciences, Northeast Ohio Medical University, Rootstown, OH;
3 Family Practice, Metrohealth Medical Center, Cleveland, OH;
4 Cleveland Clinic, Cleveland, OH
Objectives Nonalcoholic steatohepatitis (NASH) is associated
with endothelial dysfunction and an increased rate of
cardio-vascular disease (CVD) related mortality High density
lipopro-tein (HDL) protects against CVD through reverse cholesterol
transport in addition to its anti-oxidant and anti-inflammatory
functions We recently demonstrated that HDL from patients
with NASH has both reduced cholesterol efflux and
anti-oxi-dant activity and it is pro-inflammatory Here we use a heavy
water (2H2O) based metabolic labeling approach to test the
hypothesis that altered HDL proteome dynamics is involved
in HDL dysfunction in patients with NASH Methods HDL from
patients NASH and paired healthy controls (n=8/group) was
isolated by an immonoaffinity method HDL’s anti-oxidant
and inflammatory activities were quantified The HDL
pro-teome composition was analyzed by a label-free proteomics
approach A metabolic 2H2O-labeling technique was applied
to quantify turnover rates of HDL proteins Results Patients with
NASH had a higher BMI, HOMA-IR, plasma AST, ALT and
tri-glycerides As compared to controls, HDL from NASH patients
was proinflammatory with increased myeloperoxidase (MPO)
levels and accelerated oxidative activity The HDL from NASH
patients was significantly enriched with proteins involved in the
acute phase response (ceruloplasmin [Cp], serum amyloid
amy-lase, hemopexin, and the complement factor B) MPO activity
was positively associated with the pro-inflammatory index of
HDL (p=0 004) and nitration of apoAI (p=0 003) In contrast,
cholesterol efflux capacity of HDL was inversely correlated with
the MPO activity (p=0 01) and nitration of apoAI (p=0 01)
These changes were associated with the increased
degrada-tion of ApoAI and ApoAII, two major proteins of HDL involved
in cholesterol transport, but with a decreased catabolic rate
of Cp, an acute-phase HDL protein with pro-oxidant property
Alterations in ApoAI metabolism were directly correlated with
the oxidase activity and levels of Cp, suggesting that Cp could
be involved in accelerated ApoAI catabolism and HDL
dysfunc-tion in NASH Conclusions HDL dysfuncdysfunc-tion in NASH is related
to the altered dynamics of HDL proteins, and Cp may play a unique role in the increased degradation of apoAI, apoAII, apoAIV and increased production of acute phase reaction pro-teins
Disclosures:
The following people have nothing to disclose: Takhar kasumov, Kwangwon Lee, Abdullah Osme, Jaividhya Dasarathy, Ling Li, Belinda Willard, Iryna Kalinina, Srinivasan Dasarathy, Jonathan D Smith, Arthur J McCullough
1223
Impaired Adipocyte Autophagy Promotes White pocyte Browning And Protects Against Alcohol-Induced Liver Injury
Adi-Yuan Li, Hong-Min Ni, Xiaojuan Chao, Wen-Xing Ding; cology, Toxicology and Therapeutics, The University of Kansas Medical Center, Kansas City, KS
Pharma-Alcoholic liver disease (ALD) is a health problem worldwide that claims two million lives per year Accumulating evidence indi-cates that chronic alcohol consumption increases adipocyte lip-olysis resulting in decreased adipose tissue mass and increased hepatic reverse transport of free fatty acid (FFA), which contrib-utes to liver steatosis and liver injury Whether and how adi-pocyte autophagy contributes to alcohol-induced lipolysis and liver injury are not known In the present study, we generated
adipocyte-specific Atg5 knockout (KO) mice and treated these
mice together with their matched wild type mice with plus-binge alcohol model (Gao-binge model) Mass and his-tology of epididymal and retroperitoneal, and subcutaneous inguinal white adipose tissue (WAT) as well as interscapular brown adipose tissue (BAT) were compared between wild-type (WT) and KO mice Liver steatosis, inflammation and injury were evaluated, and serum lipids were also measured to study lipid turnover We found that Gao-binge alcohol increased adipocyte lipolysis and decreased fat mass in WT mice, which
chronic-were inhibited in adipocyte-specific Atg5 KO mice ingly, adipocyte-specific Atg5 KO mice had increased number
Intrigu-of multilocular lipid droplets (LDs) and mitochondria in taneous white adipocytes WAT (sWAT), suggesting that lack
subcu-of Atg5 promotes the “browning “subcu-of WAT likely via decreased mitophagy More importantly, the “browning” of WAT was further enhanced after alcohol administration As a result, adi-
pocyte-specific Atg5 KO mice had decreased serum levels of
FFA, alanine aminotransferase (ALT) and aspartate ferase (AST), and decreased hepatic triglyceride and neutro-phil infiltration compared to WT mice after Gao-binge alcohol administration In conclusion, our data indicate that inhibition
aminotrans-of adipocyte autophagy attenuates Gao-binge alcohol-induced liver injury, which might be due to increased level of “brown-ing” of WAT adipocytes via decreased autophagic removal
of mitochondria, and decreased adipocyte lipolysis via less autophagic removal of LDs
Disclosures:
The following people have nothing to disclose: Yuan Li, Hong-Min Ni, Xiaojuan Chao, Wen-Xing Ding
Trang 16Establishment of a cellular model to study
ethanol-in-duced lipophagy
Lin Wang 1,2 , Jun Zhou 1,3 , Xiaoyun Chen 1 , Bilon Khambu 1 ,
Xiao-Ming Yin 1 ; 1 Pathology and Lab Med, Indiana University Sch of
Med, Indianapolis, IN; 2 College of Animal Science and
Veteri-nary Medicine, Shandong Agricultural University, Tai’An, China;
3 Department of Minimal Invasive Surgery, Xiangya 2nd Hospital,
Changsha, China
Ethanol-induced activation of lipophagy in hepatocytes plays
an important cytoprotection against liver injury, but its
mech-anism remains to be fully determined In the present study,
ethanol-induced lipophagy was established in an
immortal-ized mouse hepatocyte line, AML-12 cells, which provided
novel information on this process This cell line was able to
metabolize ethanol and ethanol treatment elevated lipid
con-tent in these cells Cells were treated with ethanol at different
concentrations (40-160 mM) for different times (12-48 hr) We
were able to detect ethanol-induced elevation of lipid content
and the toxicity By using a series of chemical and genetic
modulators that inhibited or promoted autophagy, we could
show that ethanol-induced lipid elevation was affected by the
autophagy pathway To determine how autophagy regulated
the lipid content, we determined whether a key adaptor
mole-cule p62/SQSTM1 was involved We found that both the
auto-phagosomes marker LC3 and p62 could colocalize with lipid
droplets (LDs) in ethanol-treated AML-12 cells The lipid
accu-mulation and colocalization of autophagosomes with LDs were
significantly alleviated by p62 knockdown, indicating that p62
was an important adaptor for ethanol-induced lipophagy P62
usually binds to the ubiquitin moiety of ubiquitinated proteins
Indeed, we found that the ubiquitin signals were increased,
and they were colocalized with lipid droplets and p62 in
etha-nol-treated AML-12 cells Moreover, results of three-color
stain-ing of p62-perilipin-LDs and perilipin-Ub-LDs suggested that
colocalization of p62 with perilipin and ubiquitination of
per-ilipin occurred in this process Finally, perper-ilipin knocking-down
significantly altered ethanol-induced lipophagy, suggesting that
it could participate in this process by serving as a potential
rec-ognition target Conclusion: This study thus provides a
poten-tial mechanistic explanation to how ethanol-induced lipophagy
might occur, which may facilitate the understanding of cell’s
defense against ethanol-induced injury
Disclosures:
The following people have nothing to disclose: Lin Wang, Jun Zhou, Xiaoyun
Chen, Bilon Khambu, Xiao-Ming Yin
1225
Alcohol Exposure Differentially Regulates Immune
Responses of Liver Endothelial Cells and Hepatic Stellate
Cells to E coli Infection
Silvia Giugliano 1 , Michael Kriss 1 , Lucy M Golden-Mason 1 , Cara
Wilson 2 , Hugo R Rosen 1,3 ; 1 Division of Gastroenterology &
Hepa-tology, University of Colorado Anschutz Medical Campus, Aurora,
CO; 2 Division of Infectious Diseases, University of Colorado
Anschutz Medical Campus, Aurora, CO; 3 Denver VA Medical
Center, Denver, CO
Background: Alcohol abuse is one of the leading causes of
cirrhotic liver disease A significant percentage of cirrhotic
patients succumb to bacterial infections with infection-attributed
mortality of 30%-50% Chronic alcohol exposure increases
intestinal permeability, changing the bacterial microflora
However, the exact molecular mechanism of bacterial
trans-location and how changes in the intestinal microbiome
con-tribute to liver disease progression remains largely unknown Liver sinusoidal endothelial cells (LSECs) play an essential role
in the reticuloendothelial clearance system Abnormalities in LSEC function have been reported during experimental sepsis; however, the precise mechanisms of increased susceptibility to
bacterial infections in cirrhosis are unclear Methods: Immune
responses in primary LSECs and hepatic stellate cell line (HSCs, LX-2) were analyzed by qRT-PCR after stimulating the cells with gram-negative bacteria (10x108/ml) and/or EtOH (100 nM) Cytokine and chemokine production after bacterial exposure
was detected by ELISA Results: We demonstrate that primary
LSECs cultured with gram-negative enteric bacteria (P.copri,
B theta and E coli; 10x108/ml) for 6 hours acquire a flammatory phenotype We observed differential up-regulation
pro-in-of IL1β, IL6, CCL2 and CXCL8 Of note, the strongest immune
activation was observed after exposure of LSECs to E coli
When we exposed LX-2 cells to the same gram-negative ria we observed differential up-regulation of chemokines such
bacte-as CCL2, CXCL1 and MMP9 that are involved in monocyte
and neutrophil recruitment Also in this case, E coli infection
induced the strongest gene up-regulation To investigate how alcohol exposure would affect LSEC response to bacteria, we pre-treated LSECs for 48 hours with EtOH (100 nM) before
exposure to E.coli EtOH pre-treatment significantly reduced LSEC pro-inflammatory response to E.coli In contrast, when
LX-2 cells were pre-treated with EtOH we observed significant up-regulation of pro-inflammatory genes (IL6 and CXCL10) Furthermore, conditioned media (CM) from LSECs exposed to
E coli/EtOH when added to LX-2 cells for 6 hours induced
up-regulation of IL6 and CXCL10 Conclusions: We
demon-strate that EtOH treatment suppresses LSEC immune responses
to E coli infection and induces a pro-inflammatory state in HSCs Interestingly, CM from EtOH/E coli treated LSECs elicits
the release of soluble mediators that promote HSC activation Taken together our data suggest a model whereby LSEC act as
a central player in alcoholic liver disease where they induce immune tolerance to alcohol but also cross-talk with HSCs towards pro-inflammatory responses
Disclosures:
The following people have nothing to disclose: Silvia Giugliano, Michael Kriss, Lucy M Golden-Mason, Cara Wilson, Hugo R Rosen
1226
HBV replication increases cholesterols via upregulation
of SREBP-2/HMGCR and down-regulation of CYP-7α in alcoholic fatty liver mice
Yaqi Wang 1 , Hongwu Wang 1 , Ting Wu 1 , Danqing Hu 1 , Xingxing Weng 1 , Pei-Jer Chen 2 , Xiaoping Luo 1 , Qin Ning 1 ; 1 Tongji Hospi- tal, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; 2 Hepatitis Research Center, National Taiwan University and Hospital, Taipei, Taiwan
Backgrounds: As a high prevalence of drinking alcohol in the general population, a significant portion of chronic hepatitis
B (CHB) patients are believed to have concomitant alcoholic liver disease Cholesterol homeostasis plays an important role
in maintaining the normal physiological function of body Dysregulated cholesterol homeostasis is a characteristic of many diseases, including atherosclerosis, metabolic disorders, and numerous cancers However, the impacts on Cholesterol metabolism of alcoholic steatosis combined with HBV replica-
tion remain unknown Methods: Six-weeks old male FVB/Ncrl
mice were hydrodynamically injected of pGEM-4Z1 3HBV tor or control vector and fed with ethanol diet or control diet
vec-in random, serum ALT, AST, triglyceride, cholesterol, HBeAg and liver histology, liver cholesterol level and cholesterol
Trang 17♦ Denotes AASLD Presidential Poster of Distinction
metabolism related molecules (HMGCR,SREBP-2,CYP-7α)were
detected after 6 weeks diet treatment Results: In this model,
the characteristic of inhepatic HBV replication and alcohol fatty
liver were present HBeAg could be persistently detected in the
serum and HBcAg was positively expressed in the liver After 6
weeks ethanol diet, oil red O staining of the mice liver showed
small lipid droplets within the hepatocytes Body weight was
decreased (p<0 05), serum ALT(p<0 05) and AST (p<0 05)
were elevated significanlty And a significant increase of serum
triglyceride (p<0 05) and cholesterol level (p<0 05) in ethanol
diet feed micecompared with control diet ones Interestly, liver
cholesterol level was significantly increased in ethanol diet feed
mice with HBV replication compared with other groups These
mice also showed a significantly elevated SREBP-2/HMGCR,
an important pathway of cholesterol biosynthesis CYP-7, a key
enzyme participating in the metabolism pathway of cholesterol,
was accordingly significant decrease in these mice when
com-pared with no viral replication mice Conclusions: In this study,
we successfully constructed a murine model of alcoholic fatty
liver with HBV replication, Our data suggested a HBV
replica-tion increased hepatic cholesterol, which was evidenced due
to the up-regulation of cholesterol biosynthesis via SREBP-2/
HMGCR and down-regulation of CYP-7α
Disclosures:
Pei-Jer Chen - Advisory Committees or Review Panels: BMS, GSK, BMS, GSK,
Medigene; Consulting: Medigen, Pharmaessentia; Grant/Research Support:
Vaz-genetics; Independent Contractor: J & J; Speaking and Teaching: Roche, Roche
Qin Ning - Advisory Committees or Review Panels: ROCHE, NOVARTIS, BMS,
MSD, GSK; Consulting: ROCHE, NOVARTIS, BMS, MSD, GSK; Grant/Research
Support: ROCHE, NOVARTIS, BMS; Speaking and Teaching: ROCHE,
NOVAR-TIS, BMS, MSD, GSK
The following people have nothing to disclose: Yaqi Wang, Hongwu Wang, Ting
Wu, Danqing Hu, Xingxing Weng, Xiaoping Luo
1227
Fecal microbiota manipulation prevents dysbiosis and
alcohol-induced liver lesions in mice
Laura WRZOSEK 1 , Gladys Ferrere 1 , Williams Turpin 2 , Gabriel
Perlemuter 1,3 , Anne-Marie Cassard-Doulcier 1 ; 1 INSERM U996,
DHU Hepatinov, Université Paris-Sud, Université Paris-Saclay,
CLAMART, France; 2 Division of Gastroenterology, Zane Cohen
Centre for Digestive Diseases, Mount Sinai Hospital, Toronto, ON,
M5T 3L9, Canada; Department of Medicine, University of Toronto,
ON, M5S 1A8, Canada, Toronto, ON, Canada; 3 AP-HP,
Hepato-gastroenterology and Nutrition, Hôpital Antoine-Béclère, Clamart,
France, CLAMART, France
Background and aims: Alcoholic liver disease (ALD) is a
lead-ing cause of liver failure and mortality In humans, severe
alcoholic hepatitis is associated with key changes of
intesti-nal microbiota (IM), which influences individual sensitivity to
develop advanced ALD We used the different susceptibility
to ALD observed in two distinct animal facilities to test the
effi-ciency of two complementary strategies (fecal microbiota
trans-plantation and prebiotic treatment) to reverse dysbiosis and
prevent ALD Methods: Mice were fed alcohol in two distinct
animal facilities within a Lieber DeCarli diet Fecal microbiota
transplantation was performed with fresh feces from
alcohol-re-sistant donor mice to alcohol-sensitive receiver mice three times
a week Another group of mice received pectin during the
entire alcohol consumption period Results: Ethanol induced
steatosis and liver inflammation, which were associated with
disruption of gut homeostasis, in alcohol-sensitive, but not
alco-hol-resistant mice IM analysis showed that the proportion of
Bacteroides was specifically lower in alcohol-sensitive mice
Principal coordinate analysis showed that the IM of sensitive
and resistant mice clustered differently We targeted IM using
two different strategies to prevent alcohol-induced liver lesions: (1) pectin treatment which induced major modifications of the
IM, (2) fecal microbiota transplantation which resulted in an IM very close to that of resistant donor mice in the sensitive recipi-ent mice Both methods prevented steatosis, liver inflammation,
and restored gut homeostasis Conclusions: Manipulation of
IM can prevent alcohol-induced liver injury The IM should be considered as a new therapeutic target in ALD
Disclosures:
Gabriel Perlemuter - Advisory Committees or Review Panels: Biocodex; Board Membership: Servier; Grant/Research Support: Servier; Speaking and Teaching: Gilead, Bayer, Pileje
The following people have nothing to disclose: Laura WRZOSEK, Gladys Ferrere, Williams Turpin, Anne-Marie Cassard-Doulcier
Meng-Background & Aim: The Centers for Disease Control reported that in 2014, the overall number of alcohol-related (AR) deaths (excluding motor vehicle accidents and homicide) had reached
an all-time high and had surpassed opioid-related deaths Our aim is to investigate how AR disease has impacted our inpatient
healthcare system Methods: Using the National Inpatient
Sam-ple (NIS), we analyzed overall and annual trends in AR pitalizations and deaths from 2004 to 2013 We categorized
hos-AR hospitalizations into 1) alcohol-induced psychoses (PY) 2) alcohol dependence syndrome (DS) (acute intoxication, poly-neuropathy, cardiomyopathy, or gastritis secondary to alcohol) 3) non-dependent alcoholic syndrome (ND) and 4) alcoholic liver disease (ALD) (fatty liver disease, cirrhosis, liver damage
or acute hepatitis secondary to alcohol) Results: Overall from
2004-2013, there were 4,523,781 alcohol-related izations ALD represented the lowest proportion of AR hospi-talizations (14 1%) but the highest proportion of all AR deaths (total 96,421 deaths; ALD 37 9%, DS 28 8%, ND 22 1%,
hospital-PY, 11 2%, p < 0 05) Deaths accounted for 1 3% of AR pitalizations and were highest in ALD (5 7%) hospitalizations (Table) Although the number of ALD hospitalizations increased
hos-4 0% annually, the number ALD deaths had decreased 3 2% annually Compared to other AR cohorts, ALD had the high-est proportion of patients between the age of 45-64 (63 4%; p<0 05) 70 1% of ALD hospitalizations were male, the lowest observed (p > 0 05) proportion among AR cohorts In our sub-analysis, alcoholic hepatitis (AH) accounted for 25 3% of ALD hospitalizations with similar gender and age-specific dis-tribution, but with a lower proportion of deaths (non-AH ALD
6 5%, AH, 3 3%, p < 0 05) Conclusion: Although the number
of ALD-related hospital deaths have been declining, the portion of ALD in AR deaths remain significantly high Further efforts in addressing the effects of ALD in AR-hospitalizations may reduce mortality burden in the inpatient setting
Trang 18Aijaz Ahmed - Consulting: Bristol-Myers Squibb, Gilead Sciences Inc , Roche,
AbbVie, Shire, Janssen pharmaceuticals; Grant/Research Support: Gilead
Sci-ences Inc
The following people have nothing to disclose: George Cholankeril, Ryan B
Perumpail, Eric R Yoo, Menghan Hu, Jeffrey Nadelson
1229
Acute Alcoholic Hepatitis has Comparable Survival
fol-lowing Liver Transplantation to Other Causes of Acute
Liver Failure
George Cholankeril 1,6 , Ryan B Perumpail 2 , Menghan Hu 3 , Eric
R Yoo 4 , Andy Liu 5 , Naga P Chalasani 7 , Aijaz Ahmed 2 ; 1 Roger
Williams Medical Center, Providence, RI; 2 Division of
Gastroen-terology and Hepatology, Stanford University School of Medicine,
Stanford, CA; 3 Biostatistics, Brown University School of Public
Health, Providence, RI; 4 University of Illinois College of Medicine,
Chicago, IL; 5 Department of Medicine, California Pacific
Medi-cal Center, San Francisco, CA; 6 Department of Medicine, Boston
University School of Medicine, Boston, MA; 7 Division of
Gastroen-terology and Hepatology, Indiana University School of Medicine,
Indianapolis, IN
Background: Although liver transplantation (LT) is the
defin-itive treatment for patients with liver failure, patients with
acute alcoholic hepatitis are typically not considered
suit-able candidates due to clinical and social reasons We aim
to investigate long-term post-LT survival in the United States
(U S ) among patients with acute alcoholic hepatitis compared
to patients with other causes of acute liver failure Methods:
We conducted a cohort study using U S national data from
the United Network for Organ Sharing registry from
2005-2014 (MELD era) to evaluate long-term survival following LT
for acute alcoholic hepatitis compared to acute liver failure
including non-acetaminophen drug induced liver injury (DILI)
and acute hepatitis B Survival following LT was evaluated
with Kaplan Meier methods Results: Overall, 97 adult patients
underwent LT for acute alcoholic hepatitis, compared to 2,668
adult patients who underwent LT for acute liver failure When
compared to acute liver failure patients, acute alcoholic
hepa-titis patients were older (50 9±11 5 vs 45 8±13 8, p<0 01),
male (76 0% vs 45 9%, p<0 01), more Caucasian (78 4% vs
60 4%, p<0 01), and more likely to have undergone
hemodi-alysis in the week preceding LT (24 0% vs 16 5%, p<0 03)
Compared to liver transplants performed for acute liver failure
due to non-acetaminophen DILI (n=365) or hepatitis B (n=236)
liver transplants, acute alcoholic hepatitis demonstrated
compa-rable 1-year post-LT survival - acute alcoholic hepatitis, 82 5%;
non-acetaminophen DILI, 78 9%; and acute hepatitis B, 82 2%;
p=0 48 (Figure) Conclusions: This limited U S experience
suggests that LT performed in selected individuals with acute
alcoholic hepatitis offers an acceptable 1-year post-LT survival
Further studies are needed to better understand the utility of LT
in patients with acute alcoholic hepatitis
The following people have nothing to disclose: George Cholankeril, Ryan B Perumpail, Menghan Hu, Eric R Yoo, Andy Liu
2 NIHR Center for Liver Research and Biomedical Research Unit, University of Birmingham, Birmingham, United Kingdom; 3 Mount Sinai Medical Centre, New York, NY; 4 Department of Liver Med- icine, University Hospitals Bristol NHS Foundation Trust, Bristol, United Kingdom; 5 NIHR Nottingham Digestive Diseases Biomedi- cal Research Unit, Nottingham, United Kingdom
Alcohol and obesity synergise to increase the risk of lated mortality We examined the effect of adiposity in alco-holic hepatitis (AH) and potential underlying mechanisms Body composition in chronic liver disease was assessed by computed tomography to measure volume of adipose tissue (AT), muscle and ascites Body weight was adjusted in light
liver-re-of volume liver-re-of ascites, and patients categorised as underweight (corrected BMI (cBMI) <18 5 kg/m2), normal (18-5-24 9 kg/
m2), overweight (25-29 9 kg/m2) or obese (≥30 kg/m2) Two hundred and thirty-three patients with AH from the UK and USA provided a cohort to analyse the clinical effects of adiposity in AH Inflammatory and metabolic profiling was undertaken by proteome analysis of serum samples Analysis
of body composition showed cBMI correlated with volume of subcutaneous AT and to a lesser extent intra-abdominal AT (r2= 0 62, p<0 001 and r2=0 11 p=0 024 respectively), but did not correlate with volume of muscle (r2=0 03, p=0 215) Thus, cBMI is a reliable estimate of adiposity Overweight and obesity were common amongst patients with AH (29% and 19% respectively) Mulitvariate analysis confirmed that obesity (HR 2 22, 95%CI 1 1-4 3, p=0 022) and underweight (HR
2 38, 1 0-5 6, p=0 049) were independently associated with mortality at 3 months after admission (figure 1) Obesity was associated with greater incidence of renal failure (Chi-squared p=0 032) Proteome analysis of serum demonstrated several
Trang 19♦ Denotes AASLD Presidential Poster of Distinction
metabolic and inflammatory factors differentially expressed in
underweight and obese AH in particular adipokines,
chemo-kines and growth factors Increased adiposity is common in
patients with AH and is associated with excess morbidity and
mortality These outcomes may be mediated by differing
inflam-matory and metabolic phenotypes
Adjusted mortality at three months after admission
Disclosures:
The following people have nothing to disclose: Richard Parker, Jonathan Nahas,
Balraj Dhesi, Ashish Sinha, Antonella Ghezzi, Gene Y Im, Anne McCune,
Guruprasad P Aithal, Andrew Holt
1231
Immune regulation via the gut–liver axis plays a pivotal
role in the pathogenesis of alcohol-induced liver disease
Shunsuke Shiba, Nobuhiro Nakamoto, Ryo Aoki, Keisuke Ojiro,
Po-sung Chu, Nobuhito Taniki, Akihiro Yamaguchi, Takanori
Kanai; Department of Internal Medicine, Keio University School of
Medicine, Tokyo, Japan
Background: Excessive alcohol intake leads to the progression
of many disease ranging from gastrointestinal diseases such
as liver cirrhosis and pancreatitis to metabolic diseases In
addition to the direct effects of alcohol and its metabolites,
increased intestinal permeability and activation of hepatic
macrophages by gut microbiota-derived endotoxins through
the portal vein play a role in the pathogenesis of
alcohol-in-duced liver injury In the current study, we aimed to clarify the
immunological mechanism how alcohol-induced liver disease
progress via the gut–liver axis Method: Twenty-six patients
with alcoholic dependence admitted for the purpose of alcohol
abstinence (all male, median age: 56) were enrolled in this
study with written informed consent Blood and fecal samples
collected on admission and at 4 weeks after alcohol
cessa-tion were sequentially analyzed and compared with healthy
controls Peripheral blood mononuclear cells separated from
blood samples were analyzed for phenotype and function
using flow cytometry, MACS-sorted CD14+ monocytes were
stimulated in vitro using various toll-like receptor (TLR) ligands,
and cytokine production was evaluated using cytometric bead
assays The composition of intestinal microbiota was evaluated
using comprehensive metagenomics Results: The productivity
of inflammatory cytokines such as TNF-α and IL-6 from
periph-eral CD14+ monocytes of alcoholic patients on admission was
significantly lower than that of healthy controls The frequency
of peripheral CD14+ monocytes and the productivity of matory cytokines were significantly restored after cessation
inflam-of alcohol for 4 weeks compared with each individual’s tial value (TNF-α: pre 79 0 vs post 306 5 pg/nl, p<0 05) Cytokine production restoration was not influenced by age, ALDH1/ALDH2 gene polymorphism, amount of alcohol con-sumed, or serum transaminase level on admission Restoration was significantly poor in patients with advanced liver fibrosis even after 4 weeks of alcohol cessation Decreased diversity
ini-of intestinal microbiota due to long-term alcohol consumption
improved in most patients after alcohol cessation Conclusion:
The results suggest that peripheral monocytes in patients with alcohol dependence show diminished response to TLR4 stimula-
tion in vitro, however, tolerance to TLR ligands is reversible with
short-term alcohol cessation Further study to investigate the effects of targeting the gut-liver axis in humans may provide an undefined potential in the management of alcohol dependence.
Disclosures:
The following people have nothing to disclose: Shunsuke Shiba, Nobuhiro moto, Ryo Aoki, Keisuke Ojiro, Po-sung Chu, Nobuhito Taniki, Akihiro Yamagu- chi, Takanori Kanai
Naka-1232
Therapeutic Potential Of Liver X Receptor (LXR) In holic Liver Disease (ALD)
Alco-Monideepa Sengupta, Arindam Chatterjee, Kristine Griffett, Colin
A Flaveny, Thomas P Burris; Pharmacology and Physiology, Saint Louis University, Saint Louis, MO
The goal of our research is to determine if synthetic LXR ligands can be used to treat hepatic steatosis induced by chronic alco- hol abuse ALD is a significantly growing public health concern that is a major cause of morbidity and mortality in adults In spite of the current therapies, disease relapse frequently occurs, which raises an urgent need for the development of more effica-cious and novel therapeutics that would reverse the progression
of this disease Targeting LXR in the liver using an inverse nist has been shown to suppress hepatic lipogenesis, alter liver injury enzyme levels in the plasma and improve liver morphol-ogy in Non Alcoholic Fatty Liver Disease (NAFLD) models Due
ago-to the similarities in the pathophysiology of NAFLD and ALD,
we hypothesize that by repressing the normal function of the receptor by using synthetic inverse agonist, SR9238, we can effectively suppress or reverse the progression of ALD In order
to test the hypothesis, we performed a study where, a standard chronic ethanol diet was used where eight to ten-week-old male mice (C57BL/6J) were placed on the Lieber-DeCarli diet (Bio-Serv, Frenchtown, NJ) with 4 5% (v/v) ethanol for a total of 8 weeks Non-ethanol control-fed mice were pair-fed an equal amount of calories as their alcohol-fed counterparts The feed-ing protocol followed the guidelines established by NIAAA Two weeks into the ethanol diet, mice were administered the liver specific LXR inverse agonist SR9238 at 30mg/kg or vehi-cle control, via intraperitoneal injection, for four weeks In this experiment our results suggest that LXR inhibition significantly reduced ALD progression In another experiment, we assessed the effect of SR9238 on a “two-hit model” C57BL/6J mice were fed a high cholesterol, fructose and trans-fat diet in combi-nation with chronic ethanol treatment, in presence and absence
of SR9238 In the SR9238 treated group, we observed a nificant decrease in key liver injury enzymes including Ala-nine Transaminase(ALT), Aspartate Transaminase(AST), and Alkaline Phosphatase(ALPAMP) and a significant reduction
sig-in hepatic pro-sig-inflammatory and apoptotic markers sig-includsig-ing TNFα and TGFβ These results suggest that inhibiting LXR activ-ity may have a hepatic-protective effect in models of diet and
Trang 20alcohol induced steatosis Therefore, we predict that SR9238
would be able to attenuate liver injury by targeting the key
markers involved in the pathogenesis of ALD and subsequently
alleviate hepatic steatosis This study will also help us gain a
better understanding of the mechanisms/targets and will
fur-ther enable us to manipulate or enhance LXR function in order
to achieve novel therapeutics to treat ALD.
Disclosures:
The following people have nothing to disclose: Monideepa Sengupta, Arindam
Chatterjee, Kristine Griffett, Colin A Flaveny, Thomas P Burris
1233
Alterations of the gut microbiome in the
acute-on-chronic model of alcoholic hepatitis correlate with liver
damage, steatosis and inflammation
Patrick P Lowe 1 , Benedek Gyongyosi 1 , Abhishek
Satishchan-dran 1 , Arvin Iracheta-Vellve 1 , Aditya Ambade 1 , Doyle V Ward 2 ,
Gyongyi Szabo 1 ; 1 Department of Medicine, University of
Massa-chusetts Medical School, Worcester, MA; 2 Center for Microbiome
Research, University of Massachusetts Medical School, Worcester,
MA
Purpose: Increasingly, intestinal microbiome changes are
recognized for their profound influence on various disease
processes Several studies reveal that in alcoholic liver
ease, microbiome shifts may play a pathogenic role in
dis-ease progression In a field that relies heavily on models to
simulate human alcoholic disease, we sought to analyze the
microbiome changes in the recently published acute-on-chronic
(NIAAA) model of alcoholic hepatitis (AH) Methods: 6-8
week-old C57BL/6 female mice received 10d of 5% ethanol (EtOH)
in Lieber DeCarli liquid diet followed by EtOH gavage (5g
EtOH/kg body weight) or pair-fed diet (PF) followed by sugar
gavage Some EtOH and PF mice received a p.o cocktail of
antibiotics to decontaminate gut microbes Cecal stool DNA
was extracted and Illumina 16S sequencing was completed
using QIIME and LEfSe analysis Results: Mice treated with
antibiotics showed protection from key measures of AH
includ-ing hepatic cytokine increases, steatosis and neutrophil
infiltra-tion highlighting the importance of gut microbiota From stool
of non-antibiotic treated mice, we measured α diversity using
PD_whole_tree, chao1, observed_otus and shannon Each
quantifies diversity within a sample and compares that
sity between groups We observed no differences in α
diver-sity between PF and EtOH mice, indicating that after 10d of
alcohol, there is no suppression of α diversity Next we
exam-ined β diversity, a metric that assesses community dissimilarity,
using bray-curtis, unweighted UniFrac and weighted UniFrac
By these measures, we observed that there were no significant
community differences between PF and EtOH Finally, LEfSe
examines taxonomic changes between groups and revealed
enrichment in EtOH mice in phylum Actinobacteria, driven
by enrichment in genus Olsenella Phylum Tenericutes was
reduced in relative abundance in EtOH mice Most dramatic
was the reduction observed in phylum Verrucomicrobia driven
largely by genus Akkermansia in EtOH mice Conclusions: Our
results suggest that gut microbes affect the liver following EtOH
consumption and that alcohol alters bacterial content in the
acute-on-chronic feeding model α and β metrics did not differ
in our cohort, suggesting that the diversity within and between
EtOH and PF mice was not significantly changed However,
we do observe dynamic changes in specific taxonomies
pres-ent within each group These changes concur with published
evidence of taxonomic shifts at later time points of EtOH
admin-istration Importantly, our data suggest that the 10d
acute-on-chronic model of AH does induce microbiome changes that
impact liver inflammation and such dynamic changes may impact human AH
Uni-The following people have nothing to disclose: Patrick P Lowe, Benedek osi, Abhishek Satishchandran, Arvin Iracheta-Vellve, Aditya Ambade, Doyle V Ward
4 Pathology, University of Illinois Chicago, Chicago, IL; 5 enterology and Hepatology, Cleveland Clinic, Cleveland, OH;
Gastro-6 Pathobiology, Cleveland Clinic, Cleveland, OH; 7 Medicine, versity of Louisville, Louisville, KY; 8 Internal Medicine, University of Southwestern Texas Medical School, Dallas, TX
Uni-Purpose: Acute alcoholic hepatitis (AH) leads to high short term mortality (20-50%) In AH a disrupted gut-blood barrier can promote translocation of gut bacteria and LPS leading to mac-rophage activation Presently, there are no reliable biomarkers that predict mortality in patients with acute AH Method: 89 patients with the clinical diagnosis of AH were enrolled in four
US academic medical centers between Aug, 2013-Oct, 2015 There were 32 females and 53 males, ages 26 to 66 years Inclusion criteria were AST>80, AST/ALT ratio > 1 5, exces-sive alcohol use and no other known causes of liver disease MELD≤19 was defined as moderate AH and MELD >20 as severe AH 12 healthy subjects served as controls Results: Indi-cators of gut microbial translocation, including plasma endo-toxin (LPS), bacterial 16S DNA, soluble CD14 (sCD14) and lipoprotein binding protein (LBP), were significantly increased
in AH compared to controls Spearman correlation showed a significant correlation between MELD score, LPS and LBP levels; but not between MELD and 16S DNA or sCD14 We found a significant increase in chemokine and inflammatory cytokine production in AH patients Whole blood samples revealed a significant increase in baseline levels of TNFα, IL-1ʒ, MCP1 and
IL-8 in AH compared to healthy controls Furthermore, ex vivo
LPS stimulation elicited significantly higher secretion of TNFα, IL-1ʒ, MCP1 and IL-8 in whole blood from AH patients com-pared to controls Circulating osteopontin (OPN, a cytokine involved in neutrophil recruitment and liver regeneration) and HMGB1 (a sterile danger signal and neutrophil activator) were significantly increased in AH compared to healthy controls CD163 and CD206, scavenger receptors expressed on mac-rophages, are shed into circulation as soluble (s) CD163 and CD206 following inflammation and macrophage activation
We observed a significant increase in the levels of sCD163 and sCD206 in AH patients compared to controls Spearman correlation showed a significant positive correlation between MELD scores and OPN with sCD163 and sCD206 (but not
Trang 21♦ Denotes AASLD Presidential Poster of Distinction
HMGB1) levels in AH patients Overall mortality was 17 97%
in AH (moderate:2 27% and severe:32 61%) Serum
endo-toxin, sCD14 and OPN were the highest in non-surviving AH
patients compared to survivors Of all markers studied,
macro-phage activation markers sCD163 and sCD206 levels were
closely associated with mortality in AH Conclusion: Our study
suggests that sCD163 and sCD206 are potential markers to
differentiate severity of the disease, and together with HMGB-1
and OPN, to predict survival in AH, thus demonstrating the
importance of inflammatory macrophage activation in mortality
during AH
Disclosures:
Mack C Mitchell - Board Membership: Amygdyla Neuroscience; Stock
Share-holder: BMS, Merck, Novartis, Medtronic, Johnson & Johnson
Gyongyi Szabo - Advisory Committees or Review Panels: Alcohol Research and
Health (NIAAA), Bile Acid Council, GALAXY Project, Nature Reviews in Gastro
& Hepatology, NIH ExRNA Program, Prevent Cancer Foundation, Yale
Uni-versity Liver Center, Trek Therapeutics, UniUni-versity of Colorado Alcohol Center,
University of Southern California Liver Center, University of Pittsburgh, MSTP
EAB, Cytatx, Glympse Bio, Janssen Research & Development; Board
Member-ship: ACER, Hepatology; Consulting: Novartis, Orbimed, Roviant, Salix, Tobira,
Verlyx; Grant/Research Support: NIH-NIAAA, BMS, Gilead, Genfit, Genentech,
University of Florida, Intercept, Tobira, Takeda, Vertex
The following people have nothing to disclose: Banishree Saha, Adeyinka C
Adejumo, Karen Kodys, Patrick P Lowe, Donna Catalano, Donna Giansiracusa,
Trang Vo, Aimee Kroll, Bruce Barton, Natalia Nieto, Arthur J McCullough, Laura
E Nagy, Craig McClain
1235
Ethanol-Induced Alterations in Pancreatic and Gastric
Hormone Secretion Promotes Hepatic Steatosis
Karuna Rasineni 1,2 , Carol A Casey 1,2 , Kusum K Kharbanda 1,2 ;
1 Internal Medicine, University of Nebraska Medical Center,
Omaha, NE; 2 Research Service, VA Nebraska-Western
IowajHe-alth Care System, Omaha, NE
Purpose: Fatty liver (steatosis), the earliest and most
com-mon response of the liver to excessive ethanol consumption,
is associated with increased availability of circulating fatty
acids from adipose tissue Previous studies have revealed that
chronic ethanol exposure promotes lipolysis in adipocytes by
decreasing plasma insulin levels and disrupting
insulin-depen-dent signal transduction We further showed that circulating
levels of ghrelin, a gastric hormone that blocks insulin secretion
from pancreatic b-cells levels, is significantly elevated in
etha-nol-fed rats This led us to hypothesize that the alcohol-induced
increase in serum ghrelin levels contribute to impaired insulin
secretion from pancreatic β-cells Consequently, the reduced
insulin levels contribute to enhanced lipolysis in adipose tissue
thereby mobilizing fatty acids to the liver to promote hepatic
steatosis Methods: We used a combination of in vivo and in
vitro approaches to test our hypothesis For in vitro studies, we
utilized an insulinoma- derived cell line, INS-1E, which secretes
insulin and can metabolize ethanol These cells were treated
with 50mM ethanol and ghrelin for 48h We also used primary
rat hepatocytes that were treated with ghrelin in the presence
and absence of oleic acid for 24h to determine whether ghrelin
directly affects hepatic triglyceride accumulation For the in
vivo studies, male Wistar rats pair-fed the Lieber-DeCarli liquid
diets for 6 weeks after which a subset of rats in each group
were injected with either ghrelin receptor antagonist
([D-Lys-3] GHRP-6 or saline Results: INS-1E cells secreted 50% less
insulin following ghrelin treatment Ethanol decreased insulin
secretion by 15-20% whereas an additive effect was seen upon
combined exposure Treating hepatocytes with ghrelin did not
alter the triglyceride content However, ghrelin treatment in
the presence of oleic acid induced a 3-fold increase in the
triglyceride content In vivo treatment with the ghrelin receptor
antagonist abrogated ethanol-induced steatosis and
normal-ized serum insulin and free fatty acid levels to control levels
Conclusion: Alcohol-induced increased serum ghrelin levels contribute to impaired insulin secretion from pancreatic β cells The reduced circulating insulin levels promote adipose lipolysis causing increased fatty acid mobilization to the liver thereby exacerbating hepatic steatosis We further observed that direct ghrelin treatment promotes fat accumulation in isolated hepato-cytes These exciting data indicate that inhibition of ghrelin secretion may be a useful therapeutic option for attenuating alcohol-induced steatosis
carci-Eishiro Mizukoshi, Hidetoshi Nakagawa, Toshikatsu Tamai, suya Yamashita, Kuniaki Arai, Noriho Iida, Takeshi Terashima, Shuichi Kaneko; Kanazawa Univ, Kanazawa, Japan
Tat-Background: In previous studies, we identified alpha-fetoprotein (AFP)-derived, HLA-A24-restricted cytotoxic T lymphocyte (CTL) epitopes and some of them were attractive targets for T-cell-based immunotherapy for hepatocellular carcinoma (HCC) (Int J Cancer 118:1194-1204, 2006, Hepatology 53:1206-
1216, 2011) We also performed a phase I trial of 2 kinds of AFP-derived peptides (AFP357 and AFP403) in advanced HCC patients In this study, we examined the immune responses
in the vaccinated patients and analyzed the relationships
between the immune responses and anti-tumor effects
Meth-ods: Fifteen HLA-A24-positive patients with advanced HCC were analyzed The frequency of AFP-specific T cells induced
by vaccines was monitored by interferon-gamma linked immunospot assay The frequencies of regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs) were examined by multicolor fluorescence-activated cell sorting anal-ysis and analyzed the association with the anti-tumor effects
enzyme-In addition, we analyzed AFP-specific T cell receptors (TCRs) induced by vaccines using a newly established rapid TCR clon-
ing system‘hTEC10’ (Nat Med 19:1542-1546, 2013) Results:
Regarding the antitumor effect, 1, 8 and 6 patients showed complete response (CR), stable disease (SD) and progressive disease (PD), respectively In the patients with SD, one patient showed SD for 2 years (long SD) The frequencies of AFP-spe-cific T cells evaluated by ELISPOT assays were increased in
5 of 15 patients In the relationship between immunological and anti-tumor responses, 4 of 5 (80%) patients with AFP-spe-cific immune responses after vaccination showed CR or SD Specially, the patient with the most increased AFP-specific CTLs after vaccination showed CR The frequencies of Tregs and MDSCs before vaccinations were not associated with the anti-tumor effects However, the frequency of MDSCs after the treatment was more decreased in the patients with CR or SD than in the patients with PD Fourteen kinds of TCRs with various avidities were obtained The number of TCR repertoires is more frequent in the patients with CR or long SD The TCRs derived from the vaccinated patients with good clinical responses (CR
or long SD) had higher functions including the TCR avidities to
the peptide, cytotoxic activity, cytokine production Conclusion:
AFP-derived peptides have the possibility to provide clinical benefits in HCC patients The TCR repertoire induced by the peptide vaccines related to the clinical outcome, and efficient TCRs could be obtained from vaccinated patients with good clinical responses
Trang 22Shuichi Kaneko - Grant/Research Support: MDS, Co , Inc, Chugai Pharma , Co ,
Inc, Toray Co , Inc, Daiichi Sankyo , Co , Inc, Dainippon Sumitomo, Co , Inc,
Ajinomoto Co , Inc, Bristol Myers Squibb , Inc, Pfizer , Co , Inc, Astellas , Inc,
Takeda , Co , Inc, Otsuka„ÄÄPharmaceutical, Co , Inc, Eizai Co , Inc, Bayer
Japan, Eli lilly Japan
The following people have nothing to disclose: Eishiro Mizukoshi, Hidetoshi
Nakagawa, Toshikatsu Tamai, Tatsuya Yamashita, Kuniaki Arai, Noriho Iida,
Takeshi Terashima
1237 ♦
Phase II Studies with Refametinib or Refametinib Plus
Sorafenib in Patients with Mutant RAS Hepatocellular
Carcinoma (HCC)
Josep M Llovet 1 , Philippe Merle 2 , Karl Heinz Weiss 3 , Thomas
Yau 4 , Paul Ross 5 , Vincenzo Mazzaferro 6 , Jean-Frédéric Blanc 7 ,
Yuk T Ma 8 , Chia Jui Yen 9 , Judit Kocsis 10 , Su Pin Choo 11 ,
Wat-tana Sukeepaisarnjaroen 12 , René Gérolami 13 , Jean-Francois
Dufour 14 , Edward J Gane 15 , Baek-Yeol Ryoo 16 , Markus
Peck-Ra-dosavljevic 17 , Thong Dao 18 , Winnie Yeo 19 , Wisut Lamlertthon 20 ,
Satawat Thongsawat 21 , Michael Teufel 22 , Heiko Krissel 23 , Ho
Yeong Lim 24 ; 1 Icahn School of Medicine at Mount Sinai, New
York, NY; 2 Hôpital Hôtel Dieu, Lyon, France; 3 Liver Cancer Center
Heidelberg, Heidelberg, Germany; 4 Queen Mary Hospital, Hong
Kong, Hong Kong; 5 Guy’s & St Thomas’ NHS Foundation Trust,
London, United Kingdom; 6 The Fondazione IRCCS Istituto
Nazi-onale Tumori (National Cancer Institute) and University of Milan,
Milan, Italy; 7 Hôpital haut-Lévêque, Bordeaux, France; 8 University
Hospitals Birmingham NHS Foundation Trust, Birmingham, United
Kingdom; 9 National Cheng Kung University Hospital, Tainan,
Tai-wan; 10 Debrecen University Clinical Center, Debrecen, Hungary;
11 National Cancer Center, Singapore, Singapore; 12 Srinagarind
Hospital, Khon Kaen, Thailand; 13 Aix Marseille Université,
Mar-seille, France; 14 University Clinic for Visceral Surgery and
Medi-cine, Inselspital Bern, Bern, Switzerland; 15 New Zealand Liver &
Transplant Unit, Auckland, New Zealand; 16 Asan Medical Center,
Seoul, Korea (the Republic of); 17 Medical University of Vienna,
Vienna, Austria; 18 Caen University Hospital, Caen, France; 19
Chi-nese University of Hong Kong, Hong Kong, Hong Kong; 20
Chulab-horn Hospital, Bangkok, Thailand; 21 Maharaj Nakorn Chiang Mai
Hospital, Chiang Mai, Thailand; 22 Bayer HealthCare
Pharmaceu-ticals, Whippany, NJ; 23 Bayer AG, Berlin, Germany; 24 Samsung
Medical Center, Seoul, Korea (the Republic of)
Background: A phase II study of refametinib (a selective MEK
inhibitor) plus sorafenib in Asian patients (pts) with HCC
showed that pts with mutant RAS exhibited a robust objective
response compared to pts with wild-type RAS (Lim et al Clin
Cancer Res 20:5976, 2014) We therefore conducted two
single-arm, open-label phase II studies with refametinib alone
or in combination with sorafenib in HCC pts with mutant RAS
Pts were selected prospectively for RAS mutations using cell-free
circulating DNA (ctDNA) Methods: Pts with unresectable HCC,
Child-Pugh Class A, performance status 0-1, and no prior
sys-temic anticancer therapy for HCC (except prior sorafenib in
mono study) were eligible Plasma samples were centrally
tested using the BEAMing technology based on ctDNA to select
for KRAS/NRAS mutation Mutational status of RAS mutant pts
was analyzed by Next Generation Sequencing from ctDNA
Pts in the monotherapy trial were to be treated with
refame-tinib 50mg bid, while pts in the combination trial were to be
treated with refametinib 50mg bid and sorafenib 400 mg bid
Both trials used a 2-Stage design requiring at least 5 out of 15
responders according to mRECIST assessed by central review
for continuance to second stage Results: A total of 498 pts in
the monotherapy and 820 pts in the combination study were
enrolled with 1308 plasma samples being tested In the
mono-therapy trial the prevalence of RAS mutation was 6 5% A total
of 16 pts were treated No patient achieved a confirmed CR or
PR using mRECIST Median progression free survival (PFS) was
58 days Median time to radiological progression (TTP) was 84 days, and median overall survival (OS) was 177 days Most common TEAEs in ≥25% of pts included edema limbs, fatigue, nausea, vomiting, CPK increased, diarrhea, rash acneiform, aspartate aminotransferase increased, hypertension and rash
maculo-papular In the combination study the RAS mutation rate
was 3 3% A total of 16 pts were assigned to treatment One achieved a confirmed PR Median PFS was 46 days Median TTP was 84 days, and median OS was 427 days TEAEs in
≥25% were hypertension, fatigue, diarrhea, AST increased, CPK increased, rash acneiform, rash maculo-papular, mucositis oral, vomiting, anorexia, hypoalbuminemia and platelet count decreased Somatic aberrations of RAS mutant pts as detected
in ctDNA will be shown Conclusions: Prospective testing for
RAS family mutation using ctDNA was feasible However,
nei-ther refametinib mononei-therapy nor refametinib-sorafenib bination therapy showed sufficient efficacy to warrant further development in such a highly selected patient population Disclosures:
com-Josep M Llovet - Consulting: Bayer Pharmaceuticals, Bristol Myer Squibb, ringer-Ingelheim, Eli Lilly Pharmaceuticals, Celsion, Biocompatibles, Novartis, GlaxoSmithKline, Blueprint Medicines; Grant/Research Support: Bayer Pharma- ceuticals, Bristol Myers Squibb, Boehringer-Ingelheim
Boeh-Karl Heinz Weiss - Advisory Committees or Review Panels: Bayer Health Care, BMS; Consulting: Wilson therapeutics, GMP-O, Univar; Grant/Research Sup- port: Novartis; Speaking and Teaching: Bayer Health Care, Norgine, Orphan Europe
Thomas Yau - Consulting: Bayer, Pfizer, Novartis, Bristol-Myers Squibb Paul Ross - Advisory Committees or Review Panels: Bayer, Baxalta, Amgen; Grant/Research Support: Sanofi ; Speaking and Teaching: Bayer, Merck, Roche, Celgene, Sirtex
Jean-Frédéric Blanc - Advisory Committees or Review Panels: Lilly / Imclone, Merck Serono, Sanofi, Novartis; Speaking and Teaching: Bayer, BMS, Roche, Abbvie
Yuk T Ma - Speaking and Teaching: Bayer Jean-Francois Dufour - Advisory Committees or Review Panels: Bayer, BMS, Gil- ead, AbbVie, Novartis, Sillagen, Genfit, Intercept
Edward J Gane - Advisory Committees or Review Panels: AbbVie, Janssen, Gilead Sciences, Achillion, Merck; Speaking and Teaching: AbbVie, Gilead Sciences, Merck, Alnylam
Markus Peck-Radosavljevic - Advisory Committees or Review Panels: Bayer, cept, Gilead, Janssen, BMS, AbbVie; Consulting: Bayer, Shionogi, Boehring- er-Ingelheim, ONO Pharma, Eli Lilly, AbbVie; Grant/Research Support: Bayer, Gilead, MSD, AbbVie; Speaking and Teaching: Bayer, Gilead, MSD, AbbVie Thong Dao - Board Membership: Gilead; Grant/Research Support: Abbvie; Speaking and Teaching: Gilead, Janssen
Inter-Michael Teufel - Employment: Bayer Heiko Krissel - Employment: Bayer AG; Stock Shareholder: Bayer AG The following people have nothing to disclose: Philippe Merle, Vincenzo Maz- zaferro, Chia Jui Yen, Judit Kocsis, Su Pin Choo, Wattana Sukeepaisarnjaroen, René Gérolami, Baek-Yeol Ryoo, Winnie Yeo, Wisut Lamlertthon, Satawat Thong- sawat, Ho Yeong Lim
Background Stemness in cancer is currently of great interest as
it can be used to predict prognosis of hepatocellular carcinoma (HCC) We recently proposed an HCC classification system defined by the stem cell markers epithelial cell adhesion mole-cule (EpCAM) and α-fetoprotein (AFP) to identify HCC subtypes closely related to certain liver lineages with distinct prognosis
Trang 23♦ Denotes AASLD Presidential Poster of Distinction
(Yamashita et al, Gastroenterology 2009) Here, we
evalu-ated the utility of determining serum Dickkopf-1 (DKK-1) levels,
encoded by DKK1, a gene activated by Wnt signaling and
co-regulated with EPCAM, for the diagnosis of HCC with stem
cell features Material and Methods Patients diagnosed with
HCC at the Liver Center, Kanazawa University Hospital, Japan
from 2005 to 2012 were enrolled We measured serum DKK-1
levels using the human DKK-1 ELISA kit (Uscn Life Science Inc )
Hepatic stem cell-like (HpSC-) and mature hepatocyte-like (MH-)
HCCs were defined as previously described (Yamashita et al,
Cancer Research 2008) Clinicopathological characteristics
were determined and analyzed statistically in relation to serum
DKK-1 concentrations In addition, molecular function of DKK-1
for HCC was also evaluated using HCC xenograft model
Results The study included 357 HCC patients, 60 and 205
cases of whom had hepatitis B (HBV) or hepatitis C (HCV)
infec-tions, respectively Mean serum DKK-1 levels were 209 3 pg/
ml (range, 43 0–5556 3 pg/ml), and 54 4% of HCC patients
showed elevated DKK-1 levels (DKK-1high HCC) Serum DKK-1
levels did not correlate with those of AFP or des-γ-carboxy
pro-thrombin (DCP) HpSC-HCCs showed poor prognosis with high
serum DKK-1 levels compared with MH-HCCs who received
surgery, and DKK-1high HCCs showed a significantly high
fre-quency of portal vein invasion (p < 0 001) Among
Barce-lona Clinic Liver Cancer (BCLC) stage C patients treated with
sorafenib or hepatic arterial infusion chemotherapy using
inter-feron-α/5-FU/cisplatin, DKK-1high HCCs showed a significantly
poor prognosis compared with DKK-1low HCCs (median overall
survival 10 6 vs 13 2 months: p=0 031, and 3 4 vs 26 7
months: p=0 0005, respectively) The anti-DKK1 antibody
sig-nificantly suppressed the growth of HCC in mouse xenograft
model Conclusions Serum DKK-1 is elevated in HCC with stem
cell features The poor response of DKK-1high HCCs to sorafenib
or cytotoxic reagents warrants the needs for the development
of a novel treatment strategy against this deadly HCC subtype
Disclosures:
Shuichi Kaneko - Grant/Research Support: MDS, Co , Inc, Chugai Pharma , Co ,
Inc, Toray Co , Inc, Daiichi Sankyo , Co , Inc, Dainippon Sumitomo, Co , Inc,
Ajinomoto Co , Inc, Bristol Myers Squibb , Inc, Pfizer , Co , Inc, Astellas , Inc,
Takeda , Co , Inc, Otsuka„ÄÄPharmaceutical, Co , Inc, Eizai Co , Inc, Bayer
Japan, Eli lilly Japan
The following people have nothing to disclose: Hajime Sunagozaka, Taro
Yamashita, Tetsuro Shimakami, Kazuya Kitamura, Kuniaki Arai, Yoshio Sakai,
Tatsuya Yamashita, Eishiro Mizukoshi, Masao Honda
1239 ♦
Deregulation of the immune-related transcriptome in
Resected human Biliary Tract Cancers
Michele Ghidini 1,4 , Luciano Cascione 2 , Andrea Lampis 5 ,
Rosan-tony Pandolfo 1 , Pietro Carotenuto 1 , Francesco Trevisani 5 , Jens
Hahne 5 , Domenico Zito 5 , Vincenza Guzzardo 6 , Alessandro Zerbi 4 ,
Guido Torzilli 4 , Massimo Roncalli 4 , Lorenza Rimassa 4 , Armando
Santoro 4 , Matteo Fassan 6 , Nicola Valeri 5,3 , Chiara Braconi 1,3 ;
1 Division of Cancer Therapeutics, The Institute of Cancer Research,
Sutton, United Kingdom; 2 Institute of Oncology Research,
Bell-inzona, Switzerland; 3 The Royal Marsden NHS Trust, London,
United Kingdom; 4 Humanitas Cancer Center, Milano, Italy; 5
Divi-sion of Molecular Pathology, The Institute of Cancer Research,
Sutton, United Kingdom; 6 Internal Medicine, University of Padua,
Padua, Italy
Background: Biliary Tract Cancers (BTC) are inflammatory
can-cers with derangement of cytokines and recruitment of immune
cells Here we investigate if a transcriptomic immune profile
(IP) is activated in BTC and represents a feature of biological
behaviour Methods: RNA was extracted by tumour tissues (TT)
and matched adjacent tissues (AT) An IP of 700+
immune-re-lated transcripts was performed in TT and AT of 24 BTCs by nCounter assay CD80 expression was assessed by immunohis-tochemistry (IHC) Cox regression analysis and Kaplan Meier methods were used to correlate with Relapse Free Survival
(RFS) and derive a prognostic gene signature Results: 132
transcripts were aberrantly expressed in TT vs AT Ingenuity Pathway Analysis showed that leucocyte migration was the top function annotation De-regulation of PDZ Binding Kinase (PBK)
in TT appeared to differentiate cases with worse prognosis (nCounter p:0 08; Taqman p:0 07) We derived a list of genes whose expression was associated with RFS We observed that risk of recurrence was associated with a greater number of genes deregulated in AT in comparison to TT We shortlisted genes that maintained statistical significance at multivariate analysis (gene expression, tumour site, adjuvant chemotherapy (AC) and R0/1 resection) We observed correlation between high expression of cytotoxic T-lymphocyte antigen-4 (CTLA-4)
in the AT and RFS (p:0 0004) Cases with low CTLA4 had reduced expression of CD80, while cases with high CTLA4 had increased CD80 expression IHC expression of CD80 varied in
TT and AT No association was seen between AT CD80 sion and RFS However, CD80 expression seemed to differen-tiate prognosis in patients who did not receive AC (p:0 01), suggesting that activation of this pathway may promote relapse and may be affected from adjuvant treatment Number of CD8+ and CD4+ cells did not correlate with RFS We also derived a immuno-gene signature that could significantly dif-ferentiate relapsed cases and was an independent prognostic
expres-factor (HR 29 43, p:0 001) Conclusions: We showed that IP
is deregulated in the AT of resected BTC and correlates with relapse suggesting that deregulation of immune infiltrate in the normal tissue creates a favourable soil for cancer cell growthDisclosures:
Lorenza Rimassa - Board Membership: Eli Lilly, Merck Serono; Speaking and Teaching: Bayer, Amgen
The following people have nothing to disclose: Michele Ghidini, Luciano one, Andrea Lampis, Rosantony Pandolfo, Pietro Carotenuto, Francesco Trevis- ani, Jens Hahne, Domenico Zito, Vincenza Guzzardo, Alessandro Zerbi, Guido Torzilli, Massimo Roncalli, Armando Santoro, Matteo Fassan, Nicola Valeri, Chiara Braconi
Chan-Purpose: Evaluation of Fibroblast growth factor 19 (FGF19)
as biomarker for trial enrichment with FGFR4 inhibitors and assessment of BLU9931 (FGFR4 inhibitor) efficacy in HCC
patient derived xenograft (PDX) models Methods: A cohort of
143 fresh-frozen HCC samples was analyzed for gene sion and copy number (CN) alterations of FGF19/FGFR4 path-way components In order to explore FGF19 as biomarker, 30 HCC samples [FGF19 amplification (n=10); FGF19 overex-pression (n=10), FGF19 normal levels (n=10)] were assessed
expres-by: 1) fluorescence in situ hybridization (FISH) of 11q13 locus,
2) custom NanoString nCounter codeset for gene expression (120 genes) and CN (38 genes), and 3) FGF19 immunohis-
Trang 24tochemistry (IHC) Results were validated in an independent
cohort of 48 formalin-fixed paraffin-embedded HCC samples
For the in vivo testing of BLU9931, we analyzed 20 PDXs with
IHC and NanoString assay and selected 2 HCC PDX models
representative of high/low levels of FGF19 expression Results:
Overexpression of FGF19 (Fold change>4) was identified in
23% of HCC cases (33/143) and focal high level
amplifi-cation of FGF19 (CN≥4) in 8% (11/143) FGF 19
expres-sion in tumors with FGF19 amplification was 114 median fold
change FGF19 amplification was effectively detected by both
FISH assay (mean CN≥4; gene-to-chromosome ratio≥2) and
NanoString copy number assay (CN≥4) with high sensitivity
(100%) and accuracy (97%) NanoString gene expression
assay detected FGF19 overexpression with an accuracy of
83% Positive FGF19 staining correlated with FGF19
amplifi-cation and overexpression with an accuracy of 87% and 70%,
respectively We validate the correlation between FGF19
expression and immunostaining both in an independent HCC
cohort (R2=0 61, p=0 0003), and in PDX models (R2=0 78,
p<0 0001) In vivo, BLU9931 significantly inhibited tumor
growth vs sorafenib in a high-FGF19 PDX model (mean tumor
volume 573 ± 114 vs 1175 ± 149mm3, p=0 005) a feature
not observed in low-FGF19 PDX models Conclusions: FGF19
immunostaining accurately identifies patients with FGF19
over-expression, which represents around 25% of HCC population
These results provide the rationale for trial enrichment in the
first-in-human study testing selective FGFR4 inhibitors in patients
with advanced HCC
Disclosures:
Klaus Hoeflich - Employment: Blueprint Medicines; Stock Shareholder: Blueprint
Medicines
Chandra Miduturu - Employment: Blueprint Medicines, Inc
Qiongfang Cao - Employment: Blueprint Medicines; Stock Shareholder: Blueprint
Stephen Miller - Employment: Blueprint Medicines; Management Position:
Blue-print Medicines; Stock Shareholder: BlueBlue-print Medicines
Christoph Lengauer - Employment: Blueprint Medicines; Stock Shareholder:
Blue-print Medicines
Josep M Llovet - Consulting: Bayer Pharmaceuticals, Bristol Myer Squibb,
Boeh-ringer-Ingelheim, Eli Lilly Pharmaceuticals, Celsion, Biocompatibles, Novartis,
GlaxoSmithKline, Blueprint Medicines; Grant/Research Support: Bayer
Pharma-ceuticals, Bristol Myers Squibb, Boehringer-Ingelheim
The following people have nothing to disclose: Agrin Moeini, Roser Pinyol,
Dan-iela Sia, Robert Montal, Judit Peix, Genis Camprecios, Swan N Thung
1241 ♦
Patient Derived Organoids can be derived from biopsies
of unresectable cholangiocarcinomas and represent an useful tool for functional tumour characterization
George Vlachogiannis 3 , Elizabeth Smyth 2 , Javier Mateos 3 , Andrea Lampis 3 , Rosantony Pandolfo 1 , Pietro Carotenuto 1 , Ruwaida Begum 2 , Blanka Hezelova 2 , Zakaria Eltahir 2 , Domenico Zito 3 , Jens Hahne 3 , Chirag Nepal 5 , Mahnaz Darvish Damavandi 3 , Nick Turner 6,2 , Matteo Fassan 4 , Paul Workman 1 , Jesper B Andersen 5 , Nicola Valeri 3,2 , Chiara Braconi 1,2 ; 1 Division of Cancer Thera- peutics, The Institute of Cancer Research, Sutton, United Kingdom;
2 The Royal Marsden NHS Trust, London, United Kingdom; 3 sion of Molecular Pathology, The Institute of Cancer Research, Sutton, United Kingdom; 4 Internal Medicine, University of Padua, Padua, Italy; 5 Biotech Research and Innovation Centre, University
Divi-of Copenhagen, Copenhagen, Denmark; 6 Breakthrough Breast Cancer Research Centre, The Institute of Cancer Research, London, United Kingdom
Background: Patients Derived Organoids (PDOs) have recently emerged as organotypic cultures that recapitulate the complex three-dimensional organization of cancer PDOs have been derived from a number of tumour types; however feasibility of derivation of PDOs from cholangiocarcinoma (CCA) biopsies has not been reported to date We have developed a pro-gramme of PDOs from fresh biopsies of unresectable CCAs Here we report feasibility data as well as biological and molec-ular characterization of PDOs derived from a CT-guided liver biopsy of a patient with chemo-refractory intrahepatic CCA
(iCCA) Methods: One biopsy specimen was dissected and
embedded in matrigel and overlaid with medium optimized for selective outgrowth of tumour cells, while another two cores were fixed in formalin and embedded in paraffin Organoids were embedded in paraffin for H&E staining RNA and DNA were extracted from tumour biopsy and its matched biop-sy-PDOs Gene expression was performed through NanoString analysis, while DNA sequencing of a 200-genes panel was run with Illumina technologies High-Throughput Screening (HTS) technologies were applied to screen PDOs against 56 small drugs, and cell viability assessed by fluorimetric assays PDOs were infected with inducible RFP+ lenti-viral TripZ vec-
tors Results: Organoid culture from a single core of the biopsy
was successful and PDOs were available for functional ing within 4 weeks PDOs retained the same morphology and molecular features of the primary tumour Gene expression profiling showed that the transcriptome of PDOs recapitulated that of the primary tissue [with a Spearman r score of 0 91 for the housekeeping genes, and 0 61 for the whole transcrip-tome (p<0 0001)] iCCA PDOs were tested against >50 drugs and showed marked sensitivity (reduced cell viability >75% in comparison to DMSO) to 10 drugs, including FGFR, HSP-90, EGFR, and checkpoint inhibitors In line with these findings we observed high expression of mRNA transcripts of the EGFR, FGFR and HSP family members Stable transfection of PDOs with an inducible vector was successful and enabled microRNA
test-expression modification for functional experiments Conclusion:
PDOs can be derived from CCA biopsy specimens and provide
an attractive ex vivo assay to perform molecular
characteri-zation and functional experiments within a reasonable time frame Therefore it can represent a valuable tool to improve personalized treatment of inoperable CCA patients
Disclosures:
Paul Workman - Consulting: Vernalis; Grant/Research Support: Vernalis The following people have nothing to disclose: George Vlachogiannis, Elizabeth Smyth, Javier Mateos, Andrea Lampis, Rosantony Pandolfo, Pietro Carotenuto, Ruwaida Begum, Blanka Hezelova, Zakaria Eltahir, Domenico Zito, Jens Hahne, Chirag Nepal, Mahnaz Darvish Damavandi, Nick Turner, Matteo Fassan, Jesper
B Andersen, Nicola Valeri, Chiara Braconi
Trang 25♦ Denotes AASLD Presidential Poster of Distinction
1242 ♦
PD-L1 expression in hepatocellular carcinoma:
relation-ship with clinical and pathological features
Julien Calderaro 1,3 , Benoit Rousseau 1,2 , Giuliana amaddeo 4 ,
Mar-ion Mercey 1 , Cécile Charpy 3 , Charlotte Costentin 4 , Alain Luciani 5 ,
Elie Serge Zafrani 3 , Alexis Laurent 6,1 , Daniel Azoulay 6 , Fouad
Lafdil 1 , Jean-Michel Pawlotsky 1 ; 1 Inserm U955, Créteil, France;
2 Oncology, Henri Mondor Hospital, Créteil, France; 3 Pathology,
Henri Mondor Hospital, Créteil, France; 4 Hepatology, Henri
Mon-dor Hospital, Créteil, France; 5 Medical Imaging, Henri Mondor
Hospital, Créteil, France; 6 Hepatobiliary Surgery, Henri Mondor
Hospital, Créteil, France
The prognosis of hepatocellular carcinoma (HCC) remains
poor, with only one third of the patients eligible to curative
treatments and very limited survival benefits with the use of
sorafenib, the current standard of care for advanced disease
Recently, agents targeting the PD-L1/PD-1 immune checkpoint
were shown to display impressive antitumor activity in
vari-ous solid or hematological malignancies, including HCC, and
PD-L1 immunohistochemical expression is thought to represent a
biomarker predictive of drug sensitivity Here, we investigated
PD-L1 expression in a series of 217 HCC and correlated our
results with clinical, histological features and
immunohistochem-ical markers (PD-1, cytokeratin 19, glutamine synthetase, and
β-catenin expression) PD-L1 expression by neoplastic cells was
significantly associated with common markers of tumor
aggres-siveness (high alpha-foetoprotein (AFP) serum levels p=0 038,
satellite nodules p<0 001, macrovascular p<0 001 and
micro-vascular p<0 001 invasion, poor differentiation p<0 001) and
with the progenitor subtype of HCC (cytokeratin 19 expression
p=0 031) High PD-L1 expression by inflammatory cells of the
tumor microenvironment also correlated with high serum AFP
levels (p<0 001), macrovascular invasion (p=0 001), poor
dif-ferentiation (p=0 001), high PD-1 expression (p<0 001) and
with the so called “lymphoepithelioma-like” histological
sub-type of HCC (p=0 003) Conclusion: Our study demonstrates
that PD-L1 expression by either neoplastic or intratumoral
inflammatory cells is related to tumor aggressiveness, and
sug-gests that the response to treatments targeting the PD-L1/PD-1
immune checkpoint could be restricted to particular HCC
vari-ants Thus, enrichment of these tumor subtypes in future clinical
trials should be considered
Disclosures:
Alain Luciani - Grant/Research Support: BRACCO, GEMS; Speaking and
Teach-ing: ABBVIE
Jean-Michel Pawlotsky - Advisory Committees or Review Panels: Abbvie,
Bris-tol-Myers Squibb, Gilead, Janssen, Merck; Grant/Research Support: Gilead;
Speaking and Teaching: Bristol-Myers Squibb, Gilead, Merck, Janssen, Gilead
The following people have nothing to disclose: Julien Calderaro, Benoit
Rous-seau, Giuliana amaddeo, Marion Mercey, Cécile Charpy, Charlotte Costentin,
Elie Serge Zafrani, Alexis Laurent, Daniel Azoulay, Fouad Lafdil
1243 ♦
A Randomized, Prospective, Comparative Study about Effects and Safety of Sorafenib vs Hepatic Arterial Infu- sion Chemotherapy for Advanced Hepatocellular Carci- noma Patients with Portal Vein Tumor Thrombosis
Wang Yong Choi 2 , Woo Jin Chung 2 , Si Hyun Bae 1 , Do Seon Song 1 , Myeong Jun Song 1 , Young Seok Kim 3 , Hyung Joon Yim 4 , Young Kul Jung 4 , Sang Jun Suh 4 , Jun Yong Park 5 , Do Young Kim 5 , Seung Up Kim 5 , Sung Bum Cho 6 ; 1 Department of Internal Medicine, Seoul St Mary’s Hospital, The Catholic University of Korea, Seoul, Korea (the Republic of); 2 Department of Internal Medicine, Keimyung University School of Medicine, Daegu, Korea (the Republic of); 3 Department of Internal Medicine, Soonchunhy- ang University College of Medicine, Bucheon, Korea (the Republic of); 4 Department of Internal Medicine, Korea University, Ansan, Korea (the Republic of); 5 Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea (the Republic of);
6 Department of Internal Medicine, Hwasoon Chonnam National University Hospital, Gwangju, Korea (the Republic of)
Background/Aims: The treatment responses of advanced tocellular carcinoma (HCC) with portal vein tumor thrombo-sis (PVTT) were not acceptable and treatment modalities were limited So, we compared effects and safety of sorafenib vs
hepa-hepatic arterial infusion chemotherapy (HAIC) Methods:
We prospectively collected data of 58 advanced HCC with PVTT patients whose Child-Turcotte-Pugh (CTP) score range
5 to 7 in 6 university hospitals from January 2013 to Oct
2015 Each twenty nine patients were treated with sorafenib
or HAIC Results: 1 The mean age was 60 2±8 4 years
old and 89 7% of the patients were male Causes of HCC were HBV (67 3%), HCV (8 6%), alcohol (19 0%) and oth-ers (5 2%) CTP class A was accounted for 89 7%, modified Union for International Cancer Control (mUICC) stage IVa was
63 8%, tumor diameter >10cm was 55 2%, multiple tumor was 60 3%, infiltrative type was 56 9%, presence of main PVTT was 63 8%, median AFP value was 240 4 ng/ml 2
29 patients were enrolled to each groups Baseline teristics (sex, mean age, cause of HCC, mUICC stage, size of tumor, number of tumor, type of tumor (nodular, massive, infil-trative, diffuse), location of PVTT (main, main+branch, branch) CTP class, median value of AFP) has no significant difference between two groups 3 The median overall survival (OS) was significantly longer in HAIC group than sorafenib group (14 9 vs 7 0 months, p=0 014) The median time to progres-sion (TTP) was longer in HAIC group than sorafenib group (4 9 vs 2 2 months, p=0 009) 4 The objective response rate was 38 1% in HAIC and 4 5% in sorafenib group (p=0 003)
charac-In univariate analysis, treatment modality, main portal vein invasion, objective response, massive tumor type were signif-icant prognostic factors of overall survival (p=0 012, 0 046,
0 011, 0 041) and treatment modality, tumor number, sive tumor type were significant prognostic factors of time to progress (p=0 004, 0 043, <0 01) In multivariate analysis, objective response was a significant prognostic factor of over-all survival (p=0 048, respectively) and treatment modality was
mas-a significmas-ant prognostic fmas-actor of time to progress (p=0 016, respectively) 5 Major complications were neutropenia (6 9%, more than grade 3) and catheter-related complication (3 4%)
in HAIC group, hand-foot syndrome (20 7%) and diarrhea
(3 4%) in sorafenib group Conclusion(s): For treatment of
advanced HCC with PVTT patient, HAIC can be a valuable treatment modality like as sorafenib and more large size of study is needed
Disclosures:
Trang 26The following people have nothing to disclose: Wang Yong Choi, Woo Jin
Chung, Si Hyun Bae, Do Seon Song, Myeong Jun Song, Young Seok Kim, Hyung
Joon Yim, Young Kul Jung, Sang Jun Suh, Jun Yong Park, Do Young Kim, Seung
Up Kim, Sung Bum Cho
1244 ♦
Changing epidemiology of hepatocellular carcinoma in
Singapore – a study of 1,401 cases over 3 decades
Weiquan J Li 1 , George B Goh 1,2 , Clement Y Lin 1 , Jason P
Chang 1,2 , Chee Kiat Tan 1,2 ; 1 Gastroenterology and Hepatology,
Singapore General Hospital, Singapore, Singapore; 2 Duke-NUS
Medical School, Singapore, Singapore
Introduction The understanding and management of
hepato-cellular carcinoma (HCC) has improved worldwide However,
there is paucity of data characterizing HCC in Singapore
although it is the 4th most common cancer in males with the
2nd highest mortality in the country Aim Our study aims to
analyse the temporal trends in HCC in Singapore over the last
3 decades Methods Patients with HCC seen in the Singapore
General Hospital Department of Gastroenterology and
Hepa-tology were prospectively enrolled and stratified into 2 cohorts
(C1, 1988-1999; C2, 2000-2016) Comparison of patient
characteristics, tumor characteristics, management and survival
between the 2 cohorts was performed Survival census with the
National Registry of Births and Deaths was performed on 31st
October 2015 Results There were 1401 patients Mean age
at diagnosis of HCC for C1 and C2 was 59 8±13 3 years and
63 3±10 9 years respectively (p<0 001) More males were
affected in both cohorts (84 4% and 79 9% in C1 and C2
respectively) Hepatitis B, the predominant overall risk factor,
showed a significant decline from C1 to C2 (81 1% to 70 0%;
p<0 001), while Cryptogenic aetiology (likely NASH-related)
has risen over the same period (13 7% vs 21 5%; p<0 001)
There was no significant difference in distribution of Child-Pugh
class between cohorts Significantly more patients in C2 had
early stage disease (TNM I-II 65 4% vs 39 5%; p<0 001) than
in C1 In addition, significantly more patients in C2 than in
C1 were diagnosed through surveillance (37 3% vs 13 8%;
p<0 001), were less symptomatic (45 1% vs 76 8%; p <
0 001), and had better physical performance (ECOG 0/1
79 7% vs 57 2%; p<0 001) More C2 patients were in
Bar-celona Clinic Liver Cancer (BCLC) staging system 0 and A
(Stage 0 11 8% vs 1 1%, p<0 001; Stage A 24 7% vs 5 3%,
p<0 001) compared to C1 This translated into significantly
higher median survival in C2 compared to C1 patients (7 6 vs
3 4 months; p<0 001) Conclusion Over the past 3 decades,
Hepatitis B has declined significantly as an aetiology of HCC
in lieu of cryptogenic/NASH Surveillance is key to early
diag-nosis with better treatment options and hence significantly
improved survival
Disclosures:
The following people have nothing to disclose: Weiquan J Li, George B Goh,
Clement Y Lin, Jason P Chang, Chee Kiat Tan
1245 ♦
A novel serum biomarker, clusterin, could be an early
predictor of response to sorafenib
Takehisa Watanabe 2 , Satoshi Narahara 2,1 , Motohiko Tanaka 2 ,
Yutaka Sasaki 2 ; 1 JCHO Kumamoto General Hospital, Kumamoto,
Japan; 2 Gastroenterology and Hepatology, Graduate School of
Medical Sciences, Kumamoto University, Kumamoto, Japan
Background: Sorafenib, a multikinase inhibitor, is a standard
treatment for advanced hepatocellular carcinoma (HCC) To
predict sorafenib effect at an early stage of the treatment would
be greatly advantageous for clinical setting to continue the
current treatment or shift to other treatment Methods: One
hundred and six patients, who received sorafenib treatment
in Kumamoto University Hospital from June 2008 to March
2015, have been enrolled in this study Tumor response at three months after starting the treatment was evaluated with modified RECIST, and the patients were divided into two groups, PD as Non-Responder (NR, 31 cases), and the others
as Responder (R, 75 cases) To analyze the change in serum proteins comprehensively, proteome array was employed with five paired serum samples (pre-treatment, at one month of after starting the treatment) from each group, leading to validation
of the findings with cohort group Written informed consent
was obtained from all the participants Result: Because the
pilot proteome study demonstrated that expression of twenty proteins was significantly changed between pre-treatment and one month after starting the treatment, these proteins were pro-ceeded to ELISA using the corresponding Abs to confirm and determine the changing ratio of each candidate protein with other 20 cases in each group Finally, we validated the con-tribution of each candidate protein to the therapeutic effect of sorafenib As results, we found clusterin (CLU), an apoptosis related chaperone protein, as a significant serum biomarker
to extract NR Positive rate of change in serum CLU level in the first one month after starting the treatment (CLUp) was sig-nificantly related to NR (p=0 004) Cox proportional hazards regression analysis revealed CLUp significantly contribute to disease free survival (DFS) (HR=2 66; p=0 005) Moreover, to improve the prediction rate of sorafenib effect, we performed multivariate analysis with CLUp and additional clinical param-eters Consequently, CLUp, “AFP>10ng/dl, BUN>20mg/dl at starting of sorefenib” were extracted as significant contributing factors to NR (HR=50; p=0 017, HR=41 6; p=0 049, HR=40; p=0 040, respectively) When we proposed NR-index, using the number of positive factors in CLUp, “AFP>10ng/dl” and
“BUN>20mg/dl”, based on the regression analysis, dex=2 or 3” is significantly related to NR (p<0 001) Cox hazard analysis has also revealed that NR-index was signifi-
“NR-in-cantly related to DFS (HR=3 11, p=0 001) Conclusion: The
present findings indicate that increase in serum CLU level and the NR-index might be useful indexes to predict sorafenib effect
at an early stage after starting the treatment Disclosures:
The following people have nothing to disclose: Takehisa Watanabe, Satoshi Narahara, Motohiko Tanaka, Yutaka Sasaki
1246 ♦
Geographic variation and provider characteristics affect receipt of curative therapy in early stage hepatocellular carcinoma: a Veteran cohort study
Marina Serper 1,2 , David E Kaplan 1,2 , Tamar H Taddei 3,4 ; 1 sion of Gastroenterology and Hepatology, Hospital of the Uni- versity of Pennsylvania, Philadelphia, PA; 2 Corporal Michael J Crescenz VA Medical Center, Philadelphia, PA; 3 Department of Medicine, Digestive Diseases, Yale University School of Medi- cine, New Haven, CT; 4 VA Connecticut Healthcare System, West Haven, CT
Divi-Background and Aims: Curative therapy for early stage tocellular carcinoma (HCC) is underutilized in US populations
hepa-In a large national cohort of Veterans with HCC, we examined the prevalence and VA facility- and provider-level factors asso-ciated with receipt of curative therapy, and its effect on overall
survival among patients with early stage HCC Methods: From
2008-2010, Veterans with newly diagnosed HCC were tified from a national database and followed through 2014
Trang 27iden-♦ Denotes AASLD Presidential Poster of Distinction
Treatment modalities, specialist evaluation and clinical
out-comes were confirmed with manual chart review Curative
ther-apy was defined as resection or local ablation in the absence
of transplantation Transplantation was confirmed using the
UNOS STAR file Chi-squared tests, multivariable logistic and
Cox models evaluated associations between exposures and
outcomes Results: Among the 3988 Veterans with HCC, 28%
(n=1,127) had a single lesion less than 5 cm and were
Barce-lona Clinic Liver Cancer Stages 0 or A Among the early stage
patients, 27% had curative therapy as the first treatment (6%
resection, 21% local ablation), 6% had sorafenib, and 15%
had no therapy; 52% were treated with trans-arterial therapy;
5% underwent transplantation overall, and 7% of those with
trans-arterial therapy had transplantation In a multivariable
model adjusted for age, race, Child-Pugh class, and
Charl-son-Deyo comorbidity, factors associated with higher likelihood
of curative therapy were multi-disciplinary tumor board (OR
1 6, 95% CI 1 2-2 2), and receiving care in the North Atlantic
region (OR 1 7, 95% CI 1 2-2 4) Receipt of HCC care from
gastroenterology (OR 0 60, 95% CI 0 44-0 78) and oncology
(OR 0 55, 95% CI 0 39-0 76) specialists and Child-Pugh score
≥B (OR 0 59, 95% CI 0 43-0 80) were associated with lower
likelihood of curative therapy; hepatology care and academic
affiliation had no effect Patients reviewed at tumor board
were more likely to be seen by surgical specialists (50% with
tumor board; 34% without tumor board, p<0 001) The overall
median survival was 2 7 years (IQR:1 3-5 1) Curative therapy
(HR 0 49, 95% 0 40-0 60) was associated with a greater
sur-vival benefit than trans-arterial therapy (HR 0 60, 95% 0
.50-0 72) while oncology care in early stage HCC was associated
with worse overall survival (HR 1 3, 95% CI: 1 1-1 6)
Con-clusions: Curative HCC therapy conferred a greater survival
benefit than trans-arterial therapy, however, was under-utilized
in early stage HCC Significant unexplained geographic
vari-ation was noted in care delivery patterns Receipt of curative
therapy was associated with the type of managing specialist
provider and multi-disciplinary tumor board
ALBI and PALBI Grade Predict Survival for HCC across
Treatment Modalities and BCLC Stages in the MELD Era
Po-Hong Liu 1,4 , Chia-Yang Hsu 4,7 , Ya-Ju Tsai 8 , Cheng-Yuan
Hsia 2,4 , Yun-Hsuan Lee 1,4 , Yi-You Chiou 3,4 , Yi-Hsiang Huang 1,5 ,
Fa-Yauh Lee 1,4 , Han-Chieh Lin 1,4 , Ming-Chih Hou 1,4 , Teh-Ia Huo 1,6 ;
1 Department of Medicine, Taipei Veterans General Hospital,
Tai-pei, Taiwan; 2 Department of Surgery, Taipei Veterans General
Hospital, Taipei, Taiwan; 3 Department of Radiology, Taipei
Vet-erans General Hospital, Taipei, Taiwan; 4 Faculty of Medicine,
National Yang-Ming University, Taipei, Taiwan; 5 Institute of
Clin-ical Medicine, National Yang-Ming University, Taipei, Taiwan;
6 Institute of Pharmacology, National Yang-Ming University, Taipei,
Taiwan; 7 Department of Internal Medicine, University of Nevada
School of Medicine, Reno, NV; 8 Renown Regional Medical
Cen-ter, Reno, NV
Background: The severity of liver dysfunction in hepatocellular
carcinoma (HCC) is often estimated with Child-Turcotte-Pugh
(CTP) classification or model for end-stage liver disease (MELD)
score We aim to investigate the performance of
albumin-bili-rubin (ALBI) and platelet-albumin-bilialbumin-bili-rubin (PALBI) grade, which
are recently reported to be simple and objective measurements
for liver reserve in HCC Methods: Between 2002 and 2014,
consecutive 3,182 HCC patients were enrolled to follow up their survival The area under receiver-operator-characteristic curve (AUC) was calculated to test the discriminatory powers
over 1-, 3- and 5-year survival Results: Significant survival
differences were found across all ALBI and PALBI grades from
one to three (all p<0 001) The majority (73%) of patients were
CTP class A Within CTP class A, ALBI revealed two tic groups while PALBI segregated three prognostic groups The PABLI grade identified three different survival groups for patients undergoing resection, ablation, and chemoemboliza-tion Both ALBI and PALBI grade were capable of discerning survival among different HCC stages PALBI grade had signifi-cantly higher AUC compared with CTP classification and ALBI grade at all time points For CTP class A patients, the PALBI grade was also associated with significantly higher AUC com-pared with ALBI grade at 1- and 3-year intervals The MELD
prognos-score has the lowest AUC compared to other systems
Conclu-sions: ALBI and PALBI grade are adequate models to assess liver dysfunction in HCC The PALBI grade is consistently better
in all patients, in patients with minimally decreased liver tion, and in patients receiving different aggressive therapies
func-Disclosures:
The following people have nothing to disclose: Po-Hong Liu, Chia-Yang Hsu, Ya-Ju Tsai, Cheng-Yuan Hsia, Yun-Hsuan Lee, Yi-You Chiou, Yi-Hsiang Huang, Fa-Yauh Lee, Han-Chieh Lin, Ming-Chih Hou, Teh-Ia Huo
1248
Year 2 serum HBV DNA detectability predicts lular carcinoma in entecavir-treated patients – a 7-year cohort study of 1,680 patients with chronic hepatitis B
hepatocel-Grace L.H Wong, Vincent W Wong, Henry Lik-Yuen Chan; tute of Digestive disease, Hong Kong, China
Insti-Background: The latest AASLD guidelines for chronic hepatitis
B (CHB) define persistent viremia as a plateau in the decline
of hepatitis B virus (HBV) DNA and/or failure to achieve tectable HBV DNA level after 96 weeks of antiviral therapy
unde-We aimed to investigate the clinical impact of persistent mia with respect to incident hepatocellular carcinoma (HCC) Methods: This was a retrospective-prospective single-institution cohort study of 1,680 CHB patients who had received at least
Trang 28vire-3 years of entecavir treatment The cumulative incidences of
HCC in patients who could and could not achieve negative
serum HBV DNA at Year 1, 2, 3, 4 and 5 were evaluated
Results: The mean (± standard deviation) age was 51±12
years; 1,195 (71 1%) patients were male and 415 (24 7%)
patients had clinical cirrhosis During 78 6±17 3 months of
fol-low-up, 150 (8 9%) patients developed HCC The numbers (%)
of patients achieving undetectable HBV DNA at Year 1, 2, 3, 4
and 5 were 1,034 (61 5%), 1,188 (70 7%), 1,246 (74 2%),
1,302 (77 5%) and 1,413 (84 1%) respectively Older age,
male gender, cirrhosis, and detectable HBV DNA at Year 2
or 3 (but not that at Year 1, 4 or 5) were independent
fac-tors associated with HCC The 7-year cumulative incidence of
HCC was 12 3% (95% confidence interval: 8 9%-15 7%) vs
8 2% (6 4%-10 0%) in patients with detectable and
undetect-able HBV DNA at Year 2 respectively (log rank P=0 004); the
corresponding incidence was 11 5% (7 9%-15 1%) vs 8 6%
(6 8%-10 4%) for HBV DNA at Year 3 respectively (P=0 05)
Patients who had detectable HBV DNA at Year 2 (n=492) but
undetectable HBV DNA at Year 3 (n=61; 12 4%) and Year 4
(n=117; 23 8%) tended to be more likely to have switched to
tenofovir disoproxil fumarate (11 5% and 8 5% respectively)
compared with those with detectable HBV DNA at both time
points (4 6% and 4 5%; P=0 028 and 0 096 respectively)
Patients who had detectable HBV DNA at Year 2 but
undetect-able HBV DNA at Year 4 had significantly lower risk of HCC
than those with detectable HBV DNA at both time points; the
7-year cumulative incidence of HCC was 9 2% (5 8%-12 6%)
vs 20 6% (12 8%-28 4%); P=0 02) Conclusions: Detectable
HBV DNA at Year 2 or Year 3 after entecavir treatment is
an independent risk factor of HCC Patients who had initially
detectable HBV DNA at Year 2 but subsequently negative HBV
DNA at Year 4 had lower risk of HCC Our findings supported
the latest AASLD guideline to define persistent viremia at the
time point of 96 weeks Further study to establish the role of
adding a second antiviral drug or switching to another drug
in lieu of continuing monotherapy in this setting is warranted
Disclosures:
Grace L H Wong - Speaking and Teaching: Echosens, Echosens, Echosens,
Echosens
Vincent W Wong - Advisory Committees or Review Panels: AbbVie, Gilead,
Janssen, Tobira; Consulting: Merck, NovaMedica; Speaking and Teaching:
Gil-ead, Echosens
Henry Lik-Yuen Chan - Advisory Committees or Review Panels: Gilead, Janssen,
Bristol-Myers Squibb, Roche, Novartis Pharmaceutical, Abbvie; Speaking and
Teaching: Echosens
1249
Ultra-deep sequencing of circulating tumor DNA tifies druggable mutations: exploring applications of a liquid biopsy in HCC
iden-Ismail Labgaa 1,7 , Carlos Villacorta-Martin 1 , Delia D’Avola 1,8 , Amanda J Craig 1 , Ashley Stueck 2 , Stephen Ward 2 , M Isabel Fiel 2 , Milind Mahajan 3 , Swan N Thung 2 , Scott L Friedman 1 , Josep M Llovet 1,4 , Celina Ang 5 , Myron E Schwartz 6 , Augusto Villanueva 1,5 ; 1 Liver Diseases, Icahn School of Medicine at Mount Sinai, New York, NY; 2 Department of Pathology, Icahn School of Medicine at Mount Sinai, New York, NY; 3 Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY; 4 HCC Translational Research Laboratory, BCLC Group, IDIBAPS, CIBEREHD, Hospital Clinic Barcelona, Barcelona, Spain; 5 Division of Hematology and Medical Oncology, Icahn School of Medicine at Mount Sinai, New York, NY; 6 Department
of Surgery, Icahn School of Medicine at Mount Sinai, New York, NY; 7 Visceral Surgery, University Hospital of Lausanne (CHUV), Lausanne, Switzerland; 8 Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBEREHD), Pam- plona, Spain
Background and aims: Cellular components of solid tumors including DNA are released to the bloodstream However, evi-dence on circulating-free DNA (cfDNA) in hepatocellular car-cinoma (HCC) is limited, particularly for druggable mutations This study aimed to identify mutated cfDNA in HCC patients by
paired comparison of tissue and plasma/serum Methods: This
pilot study analyzed 43 samples from 8 HCC patients treated with surgical resection, including multiregional tumor tissue sampling (n=22), peripheral blood mononuclear cells (PMBC, n=8), plasma (n=8) and serum (n=5) DNA was extracted from 5 mL of plasma or serum using the QIAamp Circulating Nucleic Acid Kit (Qiagen) Ultra-deep DNA sequencing of 59 relevant HCC genes (as per prevalence and druggability) was performed after targeted capture suing the SureSelect method (Agilent®) Heuristic and statistical algorithms were used to call variants in tumor tissue and paired PBMC Somatic mutations in tissue were used as a standard to assess detection performance
of liquid biopsy Results: All patients were males (8/8), with
an median age of 61 years, and an average tumor size of 4 2
cm Main background etiologies were HBV (3/8) and HCV (2/8) Median cfDNA yield was 12 ng/mL, and median frag-ment size was 171 base pairs Median sequencing coverage was 1,500X and 5,500X in tissue and plasma, respectively
In tissue, 19 somatic mutations were detected in 5/8 patients, including known HCC oncogenes and tumor suppressors such
as TP53 (3/5), CTNNB1 (2/5), NTRK3 (1/5) Some tions (e g , PIK3CA) were only detected in a subset of tissue
muta-regions For all of the 5 patients with tissue mutations, sponding mutations in plasma were detected (100%) Overall, out of the 19 somatic mutations in tissue, 14 (74%) were also present in plasma 8/19 (42%) mutations were detected with high confidence while 6/19 (26%) displayed very low allele frequencies Detected variants in plasma included both drug-
corre-gable (e g , JAK1) and sub-clonal mutations (e g , PI3KCA)
Comparison of detection rate in plasma and serum in a subset
of 4 patients (12 tissue mutations) showed similar performance Mutations in cfDNA were also found in a patient with a small
HCC (2 cm) Conclusions: Ultra-deep sequencing of cfDNA in
plasma is feasible, and confidently identifies somatic mutations
in patients with resectable HCC Mutated cfDNA was detected
in all 5 patients with mutations, and 14/19 (74%) of mutations
in tissue were also present in plasma Circulating mutations included known HCC genes, druggable and sub-clonal muta-tions Liquid biopsy is a promising non-invasive tool to interro-gate HCC genetics
Trang 29♦ Denotes AASLD Presidential Poster of Distinction
Disclosures:
Scott L Friedman - Advisory Committees or Review Panels: Pfizer Pharmaceutical;
Consulting: Conatus Pharm, Exalenz, Genfit, Exalenz Biosciences, Eli Lilly
PHar-maceuticals, Fibrogen, Boehringer Ingelheim, Nitto Corp , Immune
Therapeu-tics, Blade TherapeuTherapeu-tics, Roche/Genentech Pharmaceuticals, DeuteRx, Abbvie,
Novartis, RuiYi, Kinemed, Sanofi Aventis, Chemocentryx, Nimbus
Therapeu-tics, Bristol Myers Squibb, DS Biosciences, Sandhill Medical Devices, Galmed,
Northern Biologics, Enanta Pharmaceuticals, Regado Bioscience, Raptor
Pharma-ceuticals, Fractyl Bioscience, Merck PharmaPharma-ceuticals, Genkyotex, Ironwood
Phar-maceuticals, Glycotest, Ionis PharPhar-maceuticals, Madrigal Pharmaceuticals; Grant/
Research Support: Galectin Therapeutics, Tobira Pharm, Zafgen Pharmaceuticals;
Stock Shareholder: Angion Biomedica, Intercept Pharma
Josep M Llovet - Consulting: Bayer Pharmaceuticals, Bristol Myer Squibb,
Boeh-ringer-Ingelheim, Eli Lilly Pharmaceuticals, Celsion, Biocompatibles, Novartis,
GlaxoSmithKline, Blueprint Medicines; Grant/Research Support: Bayer
Pharma-ceuticals, Bristol Myers Squibb, Boehringer-Ingelheim
The following people have nothing to disclose: Ismail Labgaa, Carlos
Villacor-ta-Martin, Delia D’Avola, Amanda J Craig, Ashley Stueck, Stephen Ward, M
Isabel Fiel, Milind Mahajan, Swan N Thung, Celina Ang, Myron E Schwartz,
Augusto Villanueva
1250
Multidisciplinary Tumor Board Adherence to AASLD
Treatment Guidelines for Hepatocellular Carcinoma:
Does It Matter?
Asrar AlAhmadi 1 , Jonathan Umbel 2,1 , Stanley M Cohen 2,1 ,
Anthony B Post 3,2 , Richard T Lee 4,1 , Pablo Ros 5 , Christopher T
Siegel 3,2 , Pierre M Gholam 2,1 ; 1 Medicine, Case Western Reserve
University, Cleveland, OH; 2 Digestive Health Institute, University
Hospitals Case Medical Center, Cleveland, OH; 3 Transplant
Insti-tute, University Hospitals Case Medical Center, Cleveland, OH;
4 Seidman Cancer Center, University Hospitals Case Medical
Cen-ter, Cleveland, OH; 5 Radiology, University Hospitals Case
Medi-cal Center, Cleveland, OH
Background Multidisciplinary Tumor Boards (MTB) are
required by NCI-designated Comprehensive Cancer Centers
Their impact in Hepatocellular Carcinoma (HCC) is unclear
We studied overall survival (OS) of patients based on the
recommendations of our MTB and its adherence to AASLD
HCC treatment practice guidelines Methods Data were
ret-rospectively collected for all patients with newly diagnosed
HCC and were censored at time of death, last known follow-up
or transplantation OS rates were analyzed using standard
Kaplan-Meier and Cox regression model Results 312 patients
with complete follow were included Average age was 62 8
years with 77 9% males 57 7% had HCV infection Overall
51 1% of patients were treated in adherence to guidelines
This included 73 3% of BCLC A, 58 8% of BCLC D, 26 8%
of BCLC B and 25 3% of BCLC C patients Adherence was
associated with longer OS by 21 months in BCLC A (HR 1 59;
95% CI 1 26-2 02) Conversely, OS was longer by 7 months
in BCLC D patients not treated per guidelines (HR 0 413; 95%
CI 0 25-0 69) Patients with BCLC B/C mostly did not receive
treatment per guidelines and their OS was not any different
(Figure 1 ) Conclusions Adherence to AASLD HCC treatment
guidelines varied across BCLC stages and was associated with
increased overall survival in patients with BCLC stage A and no
significant impact in BCLC stages B and C Conversely, BCLC
stage D patients had an overall survival benefit from MTB non
adherence to guidelines, likely due to personalized assessment
of more agressive therapies in selected patients Our data
sug-gest that synergy between evidence based guidelines and a
personalized approach is a key strength of Multidisciplinary
Tumor Boards in Hepatocellular Carcinoma
Disclosures:
Stanley M Cohen - Advisory Committees or Review Panels: Bristol-Myers Squibb, Bristol-Myers Squibb, Bristol-Myers Squibb, Bristol-Myers Squibb; Grant/Research Support: Roche, Vertex, Indenix, Roche, Vertex, Indenix, Roche, Vertex, Indenix, Roche, Vertex, Indenix; Speaking and Teaching: Gilead, Roche, Gilead, Roche, Gilead, Roche, Gilead, Roche
Anthony B Post - Speaking and Teaching: Gilead; Stock Shareholder: Amgen Pierre M Gholam - Advisory Committees or Review Panels: bayer, Gilead, ABB- VIE; Grant/Research Support: cempra, mallinckrodt, conatus, Gilead; Speaking and Teaching: intercept, Salix, abbvie
The following people have nothing to disclose: Asrar AlAhmadi, Jonathan Umbel, Richard T Lee, Pablo Ros, Christopher T Siegel
<purpose> Recent technical advances have enabled the tification of cytotoxic T lymphocyte (CTL) epitopes in various tumor-associated antigens (TAAs) for cholangiocarcinoma However, little is known about which TAA and its epitope are the most immunogenic and useful for a cancer vaccine for chol-angiocarcinoma It is also pointed out that there is a diversity
iden-in anatomic site and genetic levels We categorized giocarcinoma into intarhepatic and extrahepatic bile duct car-cinoma, and examined the differences in expression of TAAs and immune responses <method> Twenty six HLA-A24-posi-tive patients with cholangiocarcinoma (9 intarhepatic and 17 extrahepatic bile duct carcinoma) were included in the present study We examined the expression of TAAs, which included Cyp-B, Lck, SART1, SART2, SART3, p53, MRP3, hTERT, VEGFR1, VEGFR2, Survivin, MAGE-A4, Her2, WT1, βcatenin, CEA, EpCAM, EZH2, MUC5AC, GPC3, NYESO1, KIF20A, ART4 and AKR1C3, by real-time PCR in cholangiocarcinoma cell lines and human tissues including intrahepatic, extrahe-patic bile duct and gallbladder cancer HLA-A24-restricted CTL responses to epitopes derived from these TAAs were analyzed using IFN-γenzyme-linked immunospot assay (IFN-γELISPOT assay) with peripheral blood mononuclear cells <result> The expression of Cyp-B, Lck, SART1, SART2, SART3, p53, MRP3,
Trang 30cholan-hTERT, VEGFR1, VEGFR2, Survivin, MAGE-A4, Her2, WT1,
βcatenin, CEA, EpCAM, EZH2, MUC5AC, GPC3, NYESO1,
KIF20A, ART4 and AKR1C3 were observed in
cholangiocar-cinoma cell lines and human tissues There were no significant
differences in the expression between cell lines and human
tis-sues In addition, there were no significant differences between
intrahepatic and extrahepatic bile duct carcinoma In ELISPOT
assay, immune responses against SART1, p53, MRP3,
Sur-vivin2B, MAGE-A3, MAGE-A4, Her2, WT1, βcatenin, CEA,
EpCAM, GPC3, and KIF20A were observed in more than 3
of 26 patients, suggested these TAA-derived epitopes were
immunogenic There were no significant differences in immune
responses between intrahepatic and extrahepatic bile duct
car-cinoma On the other hand, immune responses against p53,
hTERT, VEGFR1, WT1, EZH2, MUC5AC, ART4 and AKR1C3
were also observed in healthy donors Taken together with
these results, SART1, MRP3, Survivin2B, MAGEA3, MAGEA4,
Her2, βcatenin, CEA, EpCAM, GPC3 and KIF20A were
con-sidered to be immunogenic and specific for
cholangiocarci-noma <conclusion> SART1, MRP3, Survivin2B, MAGEA4,
Her2, βcatenin, CEA, EpCAM, GPC3 and KIF20A were useful
targets of immunotherapy for cholangiocarcinoma
Disclosures:
Shuichi Kaneko - Grant/Research Support: MDS, Co , Inc, Chugai Pharma , Co ,
Inc, Toray Co , Inc, Daiichi Sankyo , Co , Inc, Dainippon Sumitomo, Co , Inc,
Ajinomoto Co , Inc, Bristol Myers Squibb , Inc, Pfizer , Co , Inc, Astellas , Inc,
Takeda , Co , Inc, Otsuka„ÄÄPharmaceutical, Co , Inc, Eizai Co , Inc, Bayer
Japan, Eli lilly Japan
The following people have nothing to disclose: Akihiko Kida, Eishiro Mizukoshi,
Takuya Seike, Atsushi Yonejima, Yuki Inada, Kiichiro Kaji, Kazutoshi Yamada,
Hidetoshi Nakagawa, Takeshi Terashima, Masaaki Kitahara, Noriho Iida
1252
Usefulness of modified Japan Integrated Staging (JIS)
score using albumin-bilirubin (ALBI) grade as a
prog-nostic scoring system for hepatocellular carcinoma:
analysis with 2584 Japanese patients
Hirofumi Izumoto 1 , Atsushi Hiraoka 1 , Takashi Kumada 2 ,
Hide-nori Toyoda 2 , Toshifumi Tada 2 , Tomoyuki Ninomiya 1 , Masashi
Hirooka 3 , Yoichi Hiasa 3 , Kojiro Michitaka 1 ; 1 Gastroenterology,
Ehime Prefectural Central Hosipital, Matsuyama, Japan; 2
Gastro-enterology, Ogaki Municipal Hospital, Ogaki, Japan; 3 Ehime
Uni-versity, Toon, Japan
Aim/Background: The Child-Pugh classification (CP) has some
non-objective factors, with chronic hepatitis indistinguishable
from early liver cirrhosis We retrospectively evaluated the
efficacy of albumin-bilirubin grade (ALBI) [(log10 bilirubin
(μmol/L) x 0 66) + (Albumin (g/L) x -0 0852): Grade 1:2:3
= ≤-2 60:<-2 60 to ≤-1 39:>-1 39], which has been proposed
as a new classification for hepatic function, and compared
with CP from the point of predicting prognosis of HCC patients
in combination with Tumor Node Metastasis staging (TNM)
Materials and Method: From 2000 to 2014, 2584 nạve
HCC (69 0±9 8 years old, 1850 males, 734 females, CP
A:B:C=1871:558:155) were enrolled We modified the Japan
Integrated Staging (JIS) score using ALBI, instead of CP [named
as ALBI and TNM score (ALBI-T score: 0-5 points)] We
retro-spectively investigated predictive value for prognosis of HCC
with ALBI-T (ALBI and TNM) and JIS scores (CP and TNM)
Results: When ALBI-T and JIS scores were calculated, some of
patients with each JIS score were distinguished to larger score
of ALBI-T (Table) As a result, in comparison with corresponding
score between ALBI-T and JIS, survival curves of ALBI-T showed
better than those of JIS score [median survival time (months):
score0: 137 7 vs 97 6, P=0 021; score1: 83 2 vs 74 9,
P=0 006; score2: 53 4 vs 39 7, P=0 002; score3: 27 4 vs
15 0, P<0 001; score4: 5 0 vs 4 0, P=0 057; score5: 1 4
vs 1 0, P=0 452; respectively] (Figure) Conclusion: Modified
JIS score (ALBI-T) using ALBI may be a better total prognostic scoring system than JIS score using CP for predicting survival
of patients with HCC
Disclosures:
The following people have nothing to disclose: Hirofumi Izumoto, Atsushi oka, Takashi Kumada, Hidenori Toyoda, Toshifumi Tada, Tomoyuki Ninomiya, Masashi Hirooka, Yoichi Hiasa, Kojiro Michitaka
Hira-1253
Growth Hormone and Hepatocellular Carcinoma
Manal Hassan 1 , Reham Abdel-Wahab 1,2 , Robert A Wolff 1 , Gehan Botrus 1 , Akram Shalaby 1 , Donghui Li 1 , Harrys A Torres 3 , Jeffrey S Morris 4 , Karri Ballard 8 , Ernest Hawk 5 , Ahmed Shalaby 1 , Hesham M Amin 6 , James C Yao 1 , Renato Lenzi 1 , Jone A Joss 7 , Prasun K Jalal 7 , Saira Khaderi 7 , Sahin Lacin 1 , Ahmed O Kaseb 1 ;
1 GI Medical Oncology, The University of Texas MD Anderson cer Center, Houston, TX; 2 Clinical Oncology Department, Assiut University Hospital, Houston, TX; 3 Department of infectious dis- eases, MD Anderson Cancer Center, Houston, TX; 4 Department of Biostatistics, MD Anderson Cancer Center, Houston, TX; 5 Clinical Cancer prevention, MD Anderson Cancer Center, Houston, TX;
Can-6 Hematopathology, MD Anderson Cancer Center, Houston, TX;
7 Division of Hepatology, Surgery and Liver transplantation, Baylor college of medicine, Houston, TX; 8 Myraid Genetics Inc, Salt Lake, UT
Development of hepatocellular carcinoma (HCC) is a function
of activation or inhibition of several molecular pathways where important protein, cytokine, and interleukin synthesis and destruction can be changed An alteration in the secretion pat-tern of growth hormone (GH) and insulin-like growth factor-1 (IGF-1) has been described in chronic liver diseases Reduction
of the hepatocyte mass and impaired synthesis capacity of the damaged hepatocytes leads to a decrease in IGF-1 synthesis with subsequent negative feedback regulation and increased
GH levels We previously reported an association between IGF-1 level and HCC risk and prognosis The current popula-tion study aimed to assess the association between GH and HCC risk and prognosis With approval from The University
of Texas MD Anderson Cancer Center Institutional Review Board, 763 patients with HCC were prospectively enrolled in our ongoing population study Circulating plasma levels of GH were determined in patients with HCC with cirrhosis (n=489), HCC without cirrhosis (n=274), healthy controls (n=200), and non-HCC cirrhotic patients (n=75) On the basis of Mayo clinic values of GH in men and women, we classified HCC patients
as having high GH values (women, >3 67 μg/L; men, > 9 μg/L) or low GH values (women, ≤3 67 μg/L; men, ≤ 9 μg/L)
To identify independent risk factors for HCC with high GH, we estimated the odds ratio (OR) using logistic regression analy-
Trang 31♦ Denotes AASLD Presidential Poster of Distinction
ses In addition, hazard ratios (HR) were calculated using Cox
proportional hazards models to assess the role of GH in HCC
prognosis For both OR and HR, 95% confidence intervals
(CI) were determined We observed higher mean (±standard
error [SE]) baseline GH in patients with HCC with cirrhosis
(3 5±0 2) or without cirrhosis (2 2±0 2) than in healthy controls
(0 4±0 1; P< 001) The increase was also observed among
non-HCC cirrhotic patients (3 3± 5) Restricted analysis in the
non-cirrhotic HCC population and healthy controls showed that
those with high GH had an approximately 7-fold increased risk
of HCC development compared with those with low GH (OR
6 8, 95% CI 1 3-36 4, P= 001) In addition, high GH was an
independent risk factor for mortality; the adjusted HR for overall
survival in patients with high GH was approximately two times
higher than in patients with low GH (HR 1 8, 95% CI 1 3-2 4,
P≤ 001) In conclusion, high GH may predict HCC risk and
prognosis, especially in the absence of underlying chronic liver
disease Further validation of the IGF-1/GH axis in HCC risk
and prognosis in independent populations is highly warranted
Disclosures:
Harrys A Torres - Consulting: Janssen Pharmaceuticals, Gilead Sciences, Vertex
Pharmaceuticals; Grant/Research Support: The University of Texas MD Anderson
Cancer Center, Gilead Sciences, Merck
Ernest Hawk - Advisory Committees or Review Panels: Huntsman Cancer Institute,
Kansas University Medical Center EAB, Mayo Clinic Cancer Center EAB, Ohio
State University EAB, Roswell Park Cancer Institute EAB, University of Nebraska
Medical Center EAB, UT Southwestern EAB, American Cancer Society;
Employ-ment: The University of Texas MD Anderson Cancer Center; Grant/Research
Support: NIH/NCI, NIH/NIMHD, CPRIT, Cancer Prevention Pharmaceuticals;
Speaking and Teaching: University of South Alabama Mitchell Cancer Institute,
American Association for Cancer Research, American Society of Clinical
Oncol-ogy, Johns Hopkins University, NCI-Summer Curriculum, Oncology Nursing
Soci-ety Conference, Weill Cornell Medical College, AACI-CCAF, Alliance Prevention
Committee, Alliance Prevention Committee, First Annual Meta-ECHO Meeting
James C Yao - Consulting: Novartis, Merck, Ipsen, Nektar
Saira Khaderi - Advisory Committees or Review Panels: Gilead
The following people have nothing to disclose: Manal Hassan, Reham
Abdel-Wa-hab, Robert A Wolff, Gehan Botrus, Akram Shalaby, Donghui Li, Jeffrey S
Morris, Karri Ballard, Ahmed Shalaby, Hesham M Amin, Renato Lenzi, Jone A
Joss, Prasun K Jalal, Sahin Lacin, Ahmed O Kaseb
1254
Cost-effectiveness of a risk-stratified hepatocellular
car-cinoma surveillance strategy in patients with cirrhosis
Nicolas Goossens 1,2 , Amit G Singal 6 , Lindsay Y King 7 , Karin L
Andersson 3 , Bryan C Fuchs 4 , Cecilia Besa 5 , Bachir Taouli 5 ,
Ray-mond T Chung 3 , Yujin Hoshida 1 ; 1 Liver Disease, Icahn School of
Medicine at Mount Sinai, New York, NY; 2 Gastroenterology and
Hepatology, Geneva University Hospital, Geneva, Switzerland;
3 Liver Center, Gastrointestinal Unit, Massachusetts General
Hos-pital, Boston, MA; 4 Division of Surgical Oncology, Massachusetts
General Hospital, Boston, MA; 5 Department of
Radiology/Trans-lational and Molecular Imaging Institute, Icahn School of Medicine
at Mount Sinai, New York, NY; 6 Department of Internal Medicine
& Harold C Simmons Cancer Center, UT Southwestern Medical
Center, Dallase, TX; 7 Duke University Health System, Durham, NC
Background/aim Hepatocellular carcinoma (HCC) surveillance
using biannual ultrasound is recommended in all patients with
cirrhosis; however, not all patients are at equal risk for
develop-ing HCC The aim of our study was to assess the
cost-effective-ness of HCC risk-stratified surveillance strategies, incorporating
new modalities, including molecular biomarkers and
abbre-viated contrast-enhanced MRI (AMRI, [Marks AJR 2015])
Methods A Markov model, simulating a cohort of 50-year old
compensated cirrhotic subjects, was adapted (Andersson CGH
2008) to compare risk-stratified HCC surveillance strategies
Subjects were stratified into high-, intermediate-, or low-risk
groups using molecular biomarkers (prognostic 186-gene liver
signature or epidermal growth factor [EGF] genotype)-based HCC risk scores, and subjected to different combinations of ultrasound, AMRI (cost assumed at half the technical cost of standard MRI), standard MRI, and/or no regular surveillance Quality-adjusted life year (QALY), total costs, and incremental cost-effectiveness ratio (ICER) were compared to the reference strategy: biannual ultrasound (US) for all patients (current rec-
ommendations) Results Our model recapitulated natural
his-tory estimates with a 2-year survival rate of 90% in the entire cohort for the reference strategy (biannual US for all patients) Subjects who developed HCC detected by US-based surveil-lance vs no surveillance had 3-year survival rates comparable
to reported rates (Singal PLoS Med 2014) When stratifying subjects according to the 186-gene risk score, two strategies (biannual US or biannual AMRI for high/intermediate risk patients and no surveillance in low risk patients) were partic-ularly cost-effective with ICER less than $50,000 compared
to biannual US Even after varying overall HCC incidence in sensitivity analysis (1 5 - 5 8%), the risk-stratified strategy (MRI for high/intermediate risk patients only) had the highest cost-ef-ficacy with ICER of $27,360 - $38,500 compared to biannual
US EGF genotype-based risk stratification similarly improved QALE with ICER less than $50,000, further supporting cost-ef-fectiveness of HCC risk biomarker-based stratified surveillance
Conclusions Risk-stratified personalized HCC surveillance
is cost-effective and outperforms the currently recommended surveillance strategy of biannual US uniformly applied to all patients with cirrhosis
Disclosures:
Amit G Singal - Advisory Committees or Review Panels: Bayer, Wako tics; Grant/Research Support: Gilead; Speaking and Teaching: Bayer Bryan C Fuchs - Consulting: Collagen Medical; Grant/Research Support: Nim- bus Therapeutics, Enanta Pharmaceuticals
DIagnos-Raymond T Chung - Grant/Research Support: Gilead, Mass Biologics, Abbvie, Merck, BMS
The following people have nothing to disclose: Nicolas Goossens, Lindsay Y King, Karin L Andersson, Cecilia Besa, Bachir Taouli, Yujin Hoshida
1255
Prediction of Response to Sorafenib in Hepatocellular Carcinoma: A Marker Panel by Multiple Reaction Moni- toring-Mass Spectrometry
Su Jong Yu 1 , Hyunsoo Kim 2 , Injun Yeo 2 , Jeong-Ju Yoo 1 , Dong Hyeon Lee 1 , Yuri Cho 1 , Eun Ju Cho 1 , Jeong-Hoon Lee 1 , Yoon Jun Kim 1 , Jung-Hwan Yoon 1 , Youngsoo Kim 2 ; 1 Department of Internal Medicine and Liver Research Institute, Seoul National University College of Medicine, Seoul, Korea (the Republic of); 2 Department
of Biomedical Engineering, Seoul National University, Seoul, Korea (the Republic of)
Sorafenib is the only standard treatment for advanced cellular carcinoma (HCC), but it provides modest survival ben-efits over placebo, necessitating predictive biomarkers of the response to sorafenib Serum samples were obtained before and after sorafenib treatment from 115 consecutive patients [training set (n = 65) and validation set (n = 50)] with HCC and analyzed by multiple reaction monitoring-mass spectrometry (MRM-MS) to quantify candidate biomarkers We verified a triple-marker panel to be predictive of the response to sorafenib
hepato-by MRM-MS, comprising CD5 antigen-like (CD5L), globulin J (IGJ), and galectin-3-binding protein (LG3BP), in HCC patients This panel was a significant predictor (AUROC
immuno-> 0 950) of the response to sorafenib treatment, having the best cutoff value by multivariate analysis In the training set, patients who exceeded this threshold (0 4) had significantly better overall survival (median, 21 4 months) than those with
lower values (median, 8 6 months; P = 0 001) Further, a value
Trang 32that was lower than this cutoff was an independent predictor
of poor overall survival [hazard ratio (HR), 2 728; 95%
confi-dence interval (CI), 1 312–5 672; P = 0 007] and remained
an independent predictive factor of rapid progression (HR,
2 631; 95% CI, 1 448–4 780; P = 0 002) Consequently,
when applied to the independent validation set, levels of the
cut-off value for triple-marker panel maintained their prognostic
value for poor clinical outcomes Conclusion: A discriminatory
signature that comprises a triple-marker panel independently
correlates with poorer survival and more rapid progression in
HCC patients who are treated with sorafenib These findings
provide new insights into targeted proteomics-based
biomark-ers, which might engender individualized sorafenib therapy
Disclosures:
Yoon Jun Kim - Grant/Research Support: Bristol-Myers Squibb, Roche, JW
Cre-agene, Bukwang Pharmaceuticals, Handok Pharmaceuticals, Hanmi
Pharma-ceuticals, Yuhan Pharmaceuticals; Speaking and Teaching: Bayer HealthCare
Pharmaceuticals, Gilead Science, MSD Korea, Yuhan Pharmaceuticals, Samil
Pharmaceuticals, CJ Pharmaceuticals, Bukwang Pharmaceuticals, Handok
Phar-maceuticals
The following people have nothing to disclose: Su Jong Yu, Hyunsoo Kim, Injun
Yeo, Jeong-Ju Yoo, Dong Hyeon Lee, Yuri Cho, Eun Ju Cho, Jeong-Hoon Lee,
Jung-Hwan Yoon, Youngsoo Kim
1256
Hyperlipidemia and Non-Alcoholic Steatohepatitis are
Risk Factors for Hepatocellular Carcinoma Development
in the Absence of Cirrhosis
Jennifer Phan 1 , Vivian Ng 1 , Samuel French 1 , Alan J Sheinbaum 2 ,
Steven-Huy B Han 1 ; 1 University of California Los Angeles, Los
Angeles, CA; 2 Veteran’s Affairs Los Angeles, Los Angeles, CA
Background Hepatocellular carcinoma (HCC) accounts for
~90% of primary hepatic malignancies With the exception
of chronic hepatitis B (HBV), other etiologies of chronic liver
disease typically require progression to cirrhosis before HCC
develops Recent reports have described HCC in non-cirrhotic
patients with hepatitis C (HCV) and non-alcoholic
steatohep-atitis (NASH) The aim of this study was to determine the
prevalence and to assess the predictors of HCC developing
in patients without cirrhosis and HBV Methods From
2005-2015, all hepatobiliary cancer patients seen in our Liver
Can-cer and Liver Transplant clinics were evaluated Patients were
included if >18 years old and had histologically-confirmed
HCC from liver biopsy, resection specimen, or explanted livers
Patients with HBV, non-HCC tumors, or missing paired tumor
and non-tumor liver histology were excluded Demographic
information, pertinent lab values, and co-morbid conditions
were recorded Potential predictors were evaluated using both
backward stepwise logistic regression model and
classifica-tion tree model Results Of the 1925 patients screened, 1382
patients (303 HBV, 510 non-HCC tumors, 569 without paired
tumor + non-tumor histology) were excluded Of 543 HCC
patients (409 transplant, 43 resection, 74 TACE/RFA, 17
untreated) included in the study, 29 (5 3%) patients had no
cir-rhosis histologically Of these 29 patients, 11 had HCV, 3 had
alcoholic liver disease, 3 had NASH, and 12 had cryptogenic
liver disease Logistic regression models show that patients
with hyperlipidemia and cryptogenic liver disease are more
likely to develop HCC without cirrhosis (OR -1 15 and -1 37,
p < 0 05); whereas patients with hepatitis C are more likely to
develop HCC with cirrhosis (OR 1 6, p < 0 05) Classification
tree model chose platelets, hyperlipidemia, albumin, AST, and
hepatitis C as variables to stratify patients into one of 6 groups
Patients with platelet count > 198 cells/mL and hyperlipidemia
had a 90 9% chance of developing HCC without cirrhosis
Patients with platelet count < 198 cells/mL and albumin > 4 3
g/dL had an 84 6% chance of developing HCC with cirrhosis
if hepatitis C (+), but 80 0% chance of developing HCC out cirrhosis if hepatitis C (-) The C statistic for this tree model
with-was 0 944, with sensitivity 91 1% and specificity 89 6%
Con-clusion This is the first large study to histologically confirm the development of HCC in non-cirrhotic patients without HBV Patients with hyperlipidemia and cryptogenic liver disease (most likely NASH) have increased risk of developing HCC without cirrhosis; whereas patients with hepatitis C are more likely to present with cirrhosis and HCC
The following people have nothing to disclose: Jennifer Phan, Vivian Ng, Samuel French
Background and Aims. Glypican-3 (GPC-3) is a member of the glypican family of glycosyl-phosphatidylinositol-anchored cell-surface heparan sulfate proteoglycans The expression of GPC-3 has been reported to be significantly increased in the tumor tissue of hepatocellular carcinoma (HCC), but not in other liver lesions or normal liver, which made GPC-3 an attrac-tive target for the immunotherapy of HCC GPC-3 is currently the target for most of the ongoing clinical trials using chimeric antigen receptor (CAR) T cells to treat HCC A comprehensive analysis on the tumorous expression of GPC-3 is thus essen-
tial and important for its further clinical application Methods
From December 2012 to December 2015, 1,643 consecutive patients were histologically diagnosed to have HCC either by liver biopsy (n=908) or liver resection (n=735) in our hospi-tal GPC-3 expression was analyzed by immunohistochemis-try (IHC) and were semi-quantified as negative, +, ++, and +++ Other clinical characteristics including gender, age, liver diseases that related to HCC, tumor differentiation and prolif-
eration were also respectively collected Results.GPC-3 were
detected in 1,405(85 5%) of the 1,643 assayed tissues, among which, 737 were scored as “+”, 615 as “++”, and 53 as
“+++” As shown in table-1, the expression of GPC-3 showed
no difference in HCC tissue of male and female patients High expression of GPC-3 was more commonly observed in HCC occurred in younger patients, patient with chronic hepatitis
B virus (HBV) infection or alcoholic liver disease In addition, high expression of GPC-3 was also associated with HCCs that
were poorly differentiated or actively proliferating
Conclu-sions Expression of GPC-3 can be seen in 86% of the HCC tissues whereas high expression (++ or +++) occurred in about half of the GPC-3 positive HCC GPC-3 could be an ideal ther-apeutic target of HCC
Trang 33♦ Denotes AASLD Presidential Poster of Distinction
Disclosures:
The following people have nothing to disclose: Guanghua Rong, Yongwu Li,
Ruoran Li, Yinying Lu
1258
The Role Of Serum Alphafetoprotein Determination In
HCV-Cirrhosis After Antiviral Treatment
Cristina Della Corte, Roberta D’Ambrosio, Nicole Piazza O Sed,
Angelo Sangiovanni, Alessio Aghemo, Massimo Iavarone, Stella
De Nicola, Massimo Colombo; Division of Gastroenterology and
Hepatology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore
Policlinico, University of Milan, Milan, Italy
Background and Aims: Serum alphafetoprotein (sAFP)
deter-mination has been removed from recommendation of
Inter-national Scientific Societies for the non invasive diagnosis of
hepatocellular carcinoma (HCC), because of its low sensitivity
and specificity The sensitivity and specificity of sAFP were
never defined in HCV-related cirrhosis after a sustained
virolog-ical response (SVR) to antiviral treatments To define sensitivity
and specificity of sAFP in patients with HCV-related cirrhosis
according to SVR to antiviral therapy Methods: Consecutive
patients with HCV-related cirrhosis, undergoing antiviral
ther-apy from 2007 to 2013, were enrolled at the end of antiviral
treatment SVR was defined as undetectable serum HCV-RNA
at 24 th week after treatment The diagnosis of cirrhosis was
either histological or clinically based All the patients
under-went semiannual ultrasound (US) and sAFP determination
HCC was diagnosed according to AASLD 2010
recommen-dation sAFP value was tested with commercial kits (Abbott
Inc), normal value <7 ng/ml Patients were classified into three
mutually exclusive categories 1) sAFP persistently normal (< 7
ng/dl) 2) fluctuating levels of sAFP 3) persistently elevated sAFP
levels (>7 ng/dl) Results: 192 patients, 114 (62%) males,
mean age 59 years (range 30-72) were enrolled SVR was
achieved in 98 (51%) patients De-novo HCC developed after
antiviral therapy during a mean 6 years follow-up in 6 (6%)
SVR patients and in 20 (21%) non-SVR patients (p<0 0001)
In non-SVR patients sAFP was persistently normal in 28 (30%),
fluctuating (3-54 ng/ml) in 15 (16%) and persistently elevated
(8-74 ng/ml) in 51 (54%) HCC was diagnosed in 20 (21%),
5 (18%) with persistently normal sAFP and in 15 (22%) of the
remaining patients (p =0 60) sAFP sensitivity, specificity, PPV
and NPV were 75%, 31%, 22% and 82%, respectively with an
accuracy of 40% In SVR patients sAFP was persistently normal
in 85 (87%) patients, fluctuating (3-20 ng/ml) in 3 (3%) and
persistently elevated (8-25 ng/ml) in 10 (10%) (p = 0 00001)
HCC was diagnosed in 3 (3 5%) with persistently normal AFP and in 3 (23%) of the remaining patients (p =0 03) sAFP sensitivity, specificity, PPV and NPV were 50%, 89%, 23% and 96, respectively, with an accuracy of 87% (P = 0 006) Conclusions: In HCV-related cirrhotic patients sAFP has a dif-ferent behaviour according to the achievement of SVR sAFP determination may have a role for the surveillance of cirrhotic patients who achieve SVR
Disclosures:
Massimo Iavarone - Speaking and Teaching: bayer spa, abbvie, btg, gilead Massimo Colombo - Advisory Committees or Review Panels: Gilead Sciences, Abbvie, BMS, Bayer, Merck; Speaking and Teaching: Gilead Sciences, Abbvie, BMS, Bayer, Merck, Janssen, Sanofi, Vertex
The following people have nothing to disclose: Cristina Della Corte, Roberta D’Ambrosio, Nicole Piazza O Sed, Angelo Sangiovanni, Alessio Aghemo, Stella
De Nicola
1259
Development of a Novel Circulating Tumor Cells tion System in Patients with Hepatocellular Carcinoma Using a Microcavity Array
Isola-Kazuto Takahashi 1 , Kazuya Ofuji 1 , Takuto Nosaka 1 , Yoshihiko Ozaki 1 , Hidetaka Matsuda 1 , Masahiro Ohtani 1 , Katsushi Hira- matsu 1 , Tomoyuki Nemoto 1 , Matsunaga Tatsuya 2 , Yasunari Naka- moto 1 ; 1 Second Department of internal medicine, University of Fukui, Fukuiken, Eiheijicho, Japan; 2 Hitachi Chemical Co., Ltd, Tokyo, Japan
Backgrounds and Aims: As hematogenous spread is the major route of HCC metastasis and recurrence, detection of circulating tumor cells (CTCs) has important clinical significance in HCC patients Recent reports indicated that CTCs were entrapped from unprocessed human whole blood based on differences in the size and deformability between tumor cells and other blood cells using a microcavity array (MCA) system The aim of this study was to establish a novel CTC isolation strategy by using
a MCA system and evaluate the clinical implication of CTCs in HCC patients Methods: From January 2015 to January 2016, peripheral blood samples were collected from 19 patients with HCC, 11 patients with chronic liver diseases (CLDs) without any cancers, and 7 healthy volunteers To enrich CTCs from whole blood, a microfabricated filter with a rectangular MCA was integrated with a miniaturized device The shape and porosity
of the MCA were optimized to efficiently capture tumor cells
on the microcavities under low flow resistance condition, while allowing other blood cells to effectively pass through Isolated CTCs that was stained immunochemically with cytokeratin, DAPI and CD45 was analyzed by fluorescent microscopy We defined cells characterized by immunofluorescent intensities that were positive for cytokeratin and DAPI and negative for CD45 as CTC Results: The average recovery rates of HepG2, HuH7 and PLC/PRF/5 cells using the MCA system were 66%, 76% and 99%, respectively In the samples (3mL whole blood)
of patients with HCC, the mean number of detected CTCs was
59 5±32 7 (mean±SE), and the positivity rate of patients (CTCs
> 10 cells) was 47 3% (9/19) In contrast, the mean number of CTCs was 5 8±1 3 and the positivity rate was 18 1% (2/11)
in the patients with CLDs without HCC None of 7 healthy volunteers showed positive Furthermore, the numbers of CTCs were significantly increased in the patients with vascular inva-sion and metastasis of HCC, compared to those with localized HCC (117 7±65 4 v s 7 1±2 4; p<0 05) The numbers of CTCs detected in patients with serum AFP^4ng/ml were sig-nificantly higher than those in patients with serum AFP4ng/ml (91 9±50 1 v s 3 9±2 1; p<0 05) Conclusions: The MCA system has a potential to isolate CTCs at high recovery rates
in HCC cell lines and at high sensitivity in the patients The
Trang 34numbers of CTCs are correlated with the progression of HCC,
in which more numbers of CTCs are seen in the patients with
vascular invasion and metastasis of tumor The results suggest
that the MCA system may provide a new strategy for high
detection rates of CTCs and a predictive tool for the clinical risk
of extrahepatic progression of HCC
Disclosures:
Kazuto Takahashi - Grant/Research Support: Hitachi Chemical Co , Ltd
The following people have nothing to disclose: Kazuya Ofuji, Takuto Nosaka,
Yoshihiko Ozaki, Hidetaka Matsuda, Masahiro Ohtani, Katsushi Hiramatsu,
Tomoyuki Nemoto, Matsunaga Tatsuya, Yasunari Nakamoto
1260
Hepatocellular Carcinoma in Patients Cured of Hepatitis
C Virus: Less Cirrhosis and Liver Fat than Expected
James F Crismale 1 , Erin H Doyle 1 , Chiara Rocha 2 , M Isabel Fiel 3 ,
Ashley Stueck 3 , Nicolas Goossens 1,4 , Kian Bichoupan 1 , Neal M
Patel 1 , Sara C Lewis 5 , Jasnit Makkar 5 , Ponni V Perumalswami 1 ,
Thomas Schiano 1 , Yujin Hoshida 1 , Myron E Schwartz 6 , Andrea
D Branch 1 ; 1 Division of Liver Diseases, Icahn School of Medicine
at Mount Sinai, New York, NY; 2 Recanati/Miller Transplantation
Institute, Icahn School of Medicine at Mount Sinai, New York, NY;
3 Department of Pathology, Icahn School of Medicine at Mount
Sinai, New York, NY; 4 Tisch Cancer Center, Icahn School of
Med-icine at Mount Sinai, New York, NY; 5 Department of Radiology,
Icahn School of Medicine at Mount Sinai, New York, NY; 6
Depart-ment of Surgery, Icahn School of Medicine at Mount Sinai, New
York, NY
Background: Despite achieving a sustained virological response
(SVR), patients may still develop hepatocellular carcinoma
(HCC) Data on risk factors for post-SVR HCC are needed to
focus surveillance programs on patients most likely to benefit
Aim: To assess risk factors for post-SVR HCC and to examine
alpha fetoprotein (AFP) as a potential indicator of HCC in
post-SVR patients Methods: Imaging, histological, and clinical data
of patients who developed HCC more than 12 months post-SVR
(1/2010-12/2014) were analyzed CT or MRI was utilized to
determine tumor characteristics, the degree of steatosis, and
to calculate liver volume, adjusted for ideal body weight (LV/
IBW) Estimated adequate LV was calculated using a previously
published formula (LV=[body surface area]*772) Histology of
non-tumor tissue was evaluated by blinded review using the
Knodel/Ishak system for necroinflammation and fibrosis stage,
and the Brunt scale for steatosis/steatohepatitis Multivariable
logistic regression was used to identify clinical factors
associ-ated with post-SVR HCC by comparing a subset of Cases (those
without HIV or HBV co-infection) to post-SVR non-HCC patients
enrolled in the HALT-C study AFP levels were compared
between post-SVR patients with (the Case patients) and without
(the HALT-C cohort) HCC Results: HCC was diagnosed in 38
Cases a median of 6 years post-SVR [interquartile range (IQR):
3-10] at a median age of 59 years (IQR: 56-67) The majority
(82%) were male, 44% were non-Hispanic Caucasian, and all
achieved an SVR with interferon-based therapy Only 12% had
hepatic steatosis on imaging Based on clinical/imaging data,
29% lacked cirrhosis The median LV/IBW was 22 4 mL/kg
(IQR: 19 2-27 3), similar to the median estimated adequate LV
for this population (22 5 mL/kg [IQR]: 20 9-25 0, p=0 79)
Non-tumor tissue was available for histopathological review in
16 patients, which revealed absence of cirrhosis in 6 (38%)
Most (63%) lacked any evidence of steatosis/steatohepatitis
According to multivariable logistic regression, post-SVR HCC
was associated with non-Caucasian race (p<0 01), albumin
below 3 5 g/dL (p=0 01), and platelets below 100x103/μL
(p<0 01) while controlling for smoking and AST/ALT ratio An
AFP cut-off of 4 45 had 88% specificity and 71% sensitivity for distinguishing post-SVR patients with and without HCC
Conclusions: Nearly one-third of Cases with post-SVR HCC lacked cirrhosis and most did not have steatosis Actionable biomarkers of HCC risk post-SVR are needed and may differ from biomarkers of HCC risk in patients with chronic HCV infection AFP should be explored for HCC detection in post-SVR patients (DA 031095, DK 090317)
Disclosures:
Kian Bichoupan - Consulting: Janssen, Gilead Andrea D Branch - Grant/Research Support: Gilead, Galmed The following people have nothing to disclose: James F Crismale, Erin H Doyle, Chiara Rocha, M Isabel Fiel, Ashley Stueck, Nicolas Goossens, Neal M Patel, Sara C Lewis, Jasnit Makkar, Ponni V Perumalswami, Thomas Schiano, Yujin Hoshida, Myron E Schwartz
1261
Decision Making for Surveillance Interval: Who May Extend Intervals 1 Year After Curative Treatments for Hepatocellular Carcinoma
Minjong Lee 2,1 , Sohee Oh 3 , Young Youn Cho 1 , Eun Ju Cho 1 , Jeong-Hoon Lee 1 , Su Jong Yu 1 , Nam-Joon Yi 1 , Kwang-Woong Lee 1 , Jeong Min Lee 1 , Jung-Hwan Yoon 1 , Kyung-Suk Suh 1 , Yoon Jun Kim 1 ; 1 Seoul National University Hospital, Seoul, Korea (the Republic of); 2 Department of Internal Medicine, Kangwon National University Hospital, Chuncheon, Korea (the Republic of);
3 Department of Biostatistics, Seoul Metropolitan Government Seoul National University Boramae Medical Center, Seoul, Korea (the Republic of)
Background/Aims: The guidelines recommend surveillance for hepatocellular carcinoma recurrence be performed 3-monthly during 1 year after curative treatment, and 6-monthly thereafter
in all patients This strategy did not reflect individual risk based
on patients’ tumor biology We aimed to identify patients who
can extend surveillance intervals 1 year after treatments
Meth-ods: We retrospectively analyzed 1,490 patients treated with hepatectomy/radiofrequency ablation in the Barcelona Clinic Liver Cancer stage 0/A and well-preserved liver function In patients under 3-monthly surveillance in total periods, a new model for survival was developed using multivariable analy-sis: the training (n=682)/validation cohort (n=341) Survival rates in low-risk patients by the new model were compared according to surveillance intervals 1 year after treatments: 3-monthly vs 6-monthly (n=467) after propensity score match-
ing and lead time bias correction Results: Albumin levels,
MELD score, tumor size, alpha-fetoprotein levels, and 1-year recurrence were independent factors for survival: odds ratios (OR) of 0 33, 1 12, 1 06, 1 09, and 6 99 respectively (all
P<0 01) One-year recurrence showed significantly higher OR
than other recurrence-free durations (1–2, 2–3, and >3 years,
P<0 01) A new model showed AUROC of 0 81 (the training
cohort) and 0 77 (the validation cohort) According to risk stratification based on the new model, survival rates in low-risk patients under 3-monthly surveillance were not superior to those
under 6-monthly surveillance 1 year after treatments (P=0 958)
Conclusions: A new model can identify HCC patients who may extend 6-monthly interval 1 year after curative treatments 3-monthly surveillance interval can be important for high-risk patients given high recurrence rates in 1 year Surveillance schedules can be optimized to reduce radiation hazard and cost without compromising benefits in low-risk patients Disclosures:
Jeong Min Lee - Advisory Committees or Review Panels: Bayer healthcare
Trang 35♦ Denotes AASLD Presidential Poster of Distinction
Yoon Jun Kim - Grant/Research Support: Bristol-Myers Squibb, Roche, JW
Cre-agene, Bukwang Pharmaceuticals, Handok Pharmaceuticals, Hanmi
Pharma-ceuticals, Yuhan Pharmaceuticals; Speaking and Teaching: Bayer HealthCare
Pharmaceuticals, Gilead Science, MSD Korea, Yuhan Pharmaceuticals, Samil
Pharmaceuticals, CJ Pharmaceuticals, Bukwang Pharmaceuticals, Handok
Phar-maceuticals
The following people have nothing to disclose: Minjong Lee, Sohee Oh, Young
Youn Cho, Eun Ju Cho, Jeong-Hoon Lee, Su Jong Yu, Nam-Joon Yi,
Kwang-Woong Lee, Jung-Hwan Yoon, Kyung-Suk Suh
1262
A Novel Biomarker-Based Model for the Prediction
of Response to Sorafenib and Overall Survival for
Advanced Hepatocellular Carcinoma: A Prospective
Cohort Study
Hwi Young Kim 1 , Dong Hyeon Lee 2 , Eun Ju Cho 3 , Su Jong Yu 3 ,
Yoon Jun Kim 3 , Jung-Hwan Yoon 3 , Jeong-Hoon Lee 3 ; 1 Department
of Internal Medicine, Ewha Womans University School of
Medi-cine, Seoul, Korea (the Republic of); 2 SMG-SNU Boramae Medical
Center, Seoul, Korea (the Republic of); 3 Seoul National University
Hospital, Seoul, Korea (the Republic of)
Background/aims: Sorafenib is the standard therapy for
patients with advanced hepatocellular carcinoma (HCC)
However, validated biomarkers for prediction of outcome with
sorafenib therapy are lacking The aims were to develop and
validate a biomarker-based model for predicting sorafenib
response and overall survival (OS) Methods: This prospective
cohort study included 124 consecutive patients (44
achiev-ing disease control and 80 experiencachiev-ing progression) with
Child-Pugh class A liver function who received sorafenib for
advanced HCC Potential serum biomarkers (i e , hepatocyte
growth factor [HGF], fibroblast growth factor [FGF], vascular
endothelial growth factor receptor-1, CD117, and
angiopoie-tin-2) were tested After identification of independent predictors
of the tumor response, a risk scoring system for predicting OS
was developed and 3-fold internal validation was conducted
Results: A risk scoring system was developed using six
covari-ates: etiology of underlying liver disease, fibrosis index score,
and serum levels of PIVKA-II, HGF, and FGF (Table) When
we stratified patients into group A (risk score <9), B (9≤risk
score<10), and C (risk score ≥10), this model provided good
discriminant functions on tumor response (c-index=0 869) and
12-month survival (AUC=0 825) Median OS times were 17 2
mo in group A, 11 2 mo in group B, and 6 6 mo in group
C, respectively (P<0 001) In internal validation, the model
maintained good discriminant functions on tumor response
(c-index=0 855), 12-month survival (AUC=0 828) and good
calibration functions (all P>0 05 between expected and
observed values) Conclusions: This new model including serum
FGF and HGF showed good performance in the prediction
of response to sorafenib as well as survival in patients with
advanced HCC Value of those serum markers in risk
stratifica-tion and decision of therapeutic agents needs further studies
Scoring system for the prediction of sorafenib response and
over-all survival
Abbreviations: HBV, hepatitis B virus; HCV, hepatitis C virus;
BCLC, Barcelona Clinic Liver Cancer; PIVKA-II, protein induced
by vitamin K absence/antagonist-II; FGF, fibroblast growth factor;
HGF, hepatocyte growth factor
Disclosures:
Yoon Jun Kim - Grant/Research Support: Bristol-Myers Squibb, Roche, JW agene, Bukwang Pharmaceuticals, Handok Pharmaceuticals, Hanmi Pharma- ceuticals, Yuhan Pharmaceuticals; Speaking and Teaching: Bayer HealthCare Pharmaceuticals, Gilead Science, MSD Korea, Yuhan Pharmaceuticals, Samil Pharmaceuticals, CJ Pharmaceuticals, Bukwang Pharmaceuticals, Handok Phar- maceuticals
Cre-The following people have nothing to disclose: Hwi Young Kim, Dong Hyeon Lee, Eun Ju Cho, Su Jong Yu, Jung-Hwan Yoon, Jeong-Hoon Lee
1263
Early decrease in alfafetoprotein as a prognosis factor
of response in patients with advanced hepatocellular carcinoma treated with sorafenib
Yolanda María Sanchez, Guillermo Ontanilla, Maria Teresa rer, Álvaro Giráldez, Jose Manuel Sousa, Juan Manuel Pascasio; Hospital Virgen del Rocío, Seville, Spain
Fer-Introduction: Alpha-fetoprotein (AFP) has been considered as indicator of hepatocellular carcinoma (HCC) prognosis but there is no clear evidence to date with response to Sorafenib therapy The aim of our study is to find if there is any cor-relation between AFP level and response to it Methods:76 patients with HCC treated with Sorafenib from 2008 to 2015 from a single tertiary center were retrospectively reviewed We analyzed the impact of AFP response in overall survival (OS)
in those patients with basal high levels of AFP Patients were categorized as either responders or non-responders on the basis of a decrease in AFP concentration > 20% Survival was visualized using Kaplan-Meier curves and compared by the log-rank test Spearman’s rank correlation coefficient was used
to analyze association between basal AFP and OS Results:
We analyzed 76 patients, 82% males, 90% of them were cirrhotics, caused by HCV in 54% of the cases 77% of our patients were classified as Child-Pugh A, 30 % had metastasis and 46% portal invasion Most patients were BCLC–C (59%) 54% had received previous treatments: 25 TACE, 8 resection,
1 liver transplantation and 7 ablation Median OS was 9 3 months We observed a strong inversely proportional correla-tion between basal AFP and OS, but not statistically significant (r=-0 93, p=0 34) 40% of our patients (23) were classified
as responders OS was better in these patients thani in sponders,median 19 35 months (16 53-22 18) vs 6 96 (5 27-
non-re-8 66), p=0 002 Conclusions: Basal AFP level is associated with OS A 20% decrease of AFP levels measured one month after beginning Sorafenib is associated with a statistically sig-nificant increase of overall survival The absence of 1-month AFP level decrease could be used as an early predictor of non-response to sorafenib and as a stop rule
Trang 36The following people have nothing to disclose: Yolanda María Sanchez,
Guill-ermo Ontanilla, Maria Teresa Ferrer, Álvaro Giráldez, Jose Manuel Sousa, Juan
Manuel Pascasio
1264
Non-alcoholic fatty liver disease (NAFLD) is associated
with low surveillance rate and advanced hepatocellular
carcinoma (HCC) at diagnosis
Yi Huang 1 , Michael C Wallace 2 , Leon A Adams 2 , Gerry C
Mac-Quillan 2 , George Garas 2 , Gary P Jeffrey 2 ; 1 UWA, Nedlands,
WA, Australia; 2 Sir Charles Gairdner Hospital, Perth, WA,
Aus-tralia
Early diagnosis of HCC is essential This study aimed to
inves-tigate the risk factors for non-surveillance and advanced HCC
at diagnosis and its effect on survival 243 Patients diagnosed
with HCC from 2006 to 2014 at Sir Charles Gairdner
Hospi-tal were included 71 patients had chronic hepatitis C (CHC);
31 had chronic hepatitis B (CHB); 50 had alcoholic liver
dis-ease (ALD); 43 had ALD and viral hepatitis, 33 had NAFLD;
9 had other liver disease, 6 had un-identified liver disease
121 (49 8%) patients had undergone six-monthly ultrasound
surveillance before the diagnosis of HCC The reasons for liver
imaging tests in those without ultrasound surveillance prior to
HCC diagnosis was abnormal liver function tests (17 1%);
symptoms including fatigue, weight loss, and abdominal
dis-comfort (27%); management of liver decompensations (33 3%)
and assessment of non-liver related diseases (22 5%) The
surveillance rate was significantly lower in NAFLD patients
(18 2%) than other liver diseases (CHC: 70 2%, CHB: 58 1%,
ALD: 48%, ALD and viral hepatitis: 44 2%, other: 44 4%;
p<0 001) 27 6% of NAFLD patients were aware of existing
liver disease before the diagnosis of HCC This was greatly
lower than other liver diseases (CHC: 91 4%, CHB: 83 9%,
ALD: 79 5%, ALD and viral hepatitis: 78 6%, other: 100%;
p<0 001) 36% of current drinkers underwent surveillance,
71% for ex-drinkers and 45% for non-drinkers (p<0 001) The
median time from the first suspicious imaging test to the
diag-nosis of HCC was 1 6 months (IQR: 0 2-4 2) 159 patients
had one HCC lesion at diagnosis; 33 had two; 6 had three;
45 had multifocal HCC 76 (31 2%) patients presented with
advanced HCC (multifocal HCC or HCC with total diameter
≥6cm) and 20% of these were undergoing surveillance The
surveillance rate was significantly higher in those with early
diagnosis of HCC (63%, p<0 001) 42 2% of NAFLD patients
and 39 5% of patients with both ALD and viral hepatitis had advanced HCC at diagnosis These were significantly higher than the other liver diseases (16 9-33 3%, p<0 001) Alcohol consumption (p<0 001), poor tumor differentiation (p=0 043), high platelet count (p<0 001) and high AFP (p=0 004) were also associated with advanced HCC at diagnosis The median survival of all patients was 47 months Patients who under-went surveillance had a significantly longer median survival compared to those who did not (53 vs 32 months, p<0 001)
In conclusion, lack of HCC surveillance is a major cause of advanced HCC at diagnosis and leads to decreased survival NAFLD patients had a significantly lower surveillance rate and higher risk of advanced HCC at diagnosis compared to the other causes of liver disease
Disclosures:
Leon A Adams - Patent Held/Filed: Quest diagnostics The following people have nothing to disclose: Yi Huang, Michael C Wallace, Gerry C MacQuillan, George Garas, Gary P Jeffrey
1265
Association of relative liver enhancement of PA-enhanced magnetic resonance images with histo- logic grade of hepatocellular carcinoma
Gd-EOB-DT-Young-Joo Jin, Jin-Woo Lee; Internal Medicine, Inha University Hospital, Inha University School of Medicine, Incheon, Korea (the Republic of)
Backgrounds/Aim: The association of histological grading of hepatocellular carcinoma (HCC) with enhancement degree
on Gd-EOB-DTPA (Primovist)-enhanced magnetic resonance images (MRI) has remained to be determined In this study,
we evaluated whether histologic grade of HCC can be dicted before surgery using enhancement degree of HCC on Primovist-enhanced MRI that was performed before surgery in
pre-HCC patients Methods: A total of 121 patients who underwent
curative surgical resection for HCC at our institution between January 2012 and March 2015 were analyzed Primovist-en-hanced MRI was performed in all patients before surgery Sig-nal intensities of HCC and peri-HCC areas were measured
by using a region of interest The relative intensity ratios of HCC lesion to the surrounding non-HCC area on unenhanced images (pre-contrast ratio) and those on hepatobiliary phase images (post-contrast ratio) were calculated Moreover, relative liver enhancement (RLE) ratio (post-contrast ratio/pre-contrast ratio) was also calculated Edmondson-Steiner (E-S) grading
system was used for histologic grading of HCC Results: Of
the 121 patients, Edmondson-Steiner grades I, II, III, and IV were observed in 2 (1 7%), 14 (11 6%), 54 (44 6%), and 51 (42 1%), respectively Based on the E-S grades I/II (n=16), III (n=54), and IV (n=51), mean RLE (%) were 85 5, 84 9, and
71 2, respectively, with statistical significance (p=0 01), and
based on E-S grade I-III (n=70) and IV (n=51), mean RLE (%)
were 85 1 and 71 2, respectively (p<0 01) Barcelona Clinic
Liver Cancer (BCLC) stage A (versus 0) (Odds ratio [OS] 4 38,
p=0 03) and mean RLE (OS 0 05, p<0 01) were found to be
significant predictive factors of E-S grade IV Conclusions: E-S
grade IV can be preoperatively predicted using RLE of ist-enhanced MRI in HCC patients
Primov-Disclosures:
The following people have nothing to disclose: Young-Joo Jin, Jin-Woo Lee
Trang 37♦ Denotes AASLD Presidential Poster of Distinction
1266
Quantifying the likely benefits and harms of
surveil-lance for hepatocellular carcinoma in patients with
cirrhosis
Eleanor Taylor 1 , Rebecca Jones 1 , J A Guthrie 3 , Ian A Rowe 1,2 ;
1 Liver Unit, Leeds Teaching Hospitals NHS Trust, Leeds, United
Kingdom; 2 Leeds Institute for Data Analytics, University of Leeds,
Leeds, United Kingdom; 3 Radiology, Leeds Teaching Hospitals
NHS Trust, Leeds, United Kingdom
Background Surveillance for hepatocellular carcinoma (HCC)
in patients with cirrhosis is recommended to detect early-stage
HCC The aim of this study was to provide estimates of
ben-efit and harms of surveillance for HCC using a modelling
approach Methods Model parameters were extracted from
prospective studies of patients undergoing surveillance,
stud-ies defining the diagnostic accuracy of imaging modalitstud-ies,
and outcomes of patients with HCC diagnosed in surveillance
These parameters were then modelled in a 1000 patient cohort
using the AASLD endorsed recall policy to define follow-up
testing Benefit was defined by the absolute number of deaths
prevented by surveillance and harm by the number of
addi-tional imaging tests and invasive procedures done Results For
every 1000 patients with cirrhosis 114 have HCC diagnosed
on subsequent testing following 5 years of surveillance (Table)
The estimated absolute reduction in mortality from HCC is 11
per 1000 patients In sensitivity analyses the benefits of
sur-veillance were increased by improved treatment outcomes to
a maximum of 29 per 1000 patients 166 patients have a
false positive ultrasound: 68 of these patients have additional
cross-sectional imaging and, according to the AASLD endorsed
recall policy, 56 have a liver biopsy These additional tests are
defined as “unnecessary” since they do not diagnose HCC
and would not have been required if the patient was not under
surveillance In addition, 23 patients have HCC diagnosed
beyond conventional curative criteria Conclusions The
bene-fits of HCC surveillance are balanced by harms of
unneces-sary diagnostic testing Quantification of both the benefits and
harms of surveillance provides information to clinicians and
patients to make individualized decisions about participation
in surveillance
Table Outcomes of surveillance for HCC in patients with cirrhosis
Disclosures:
J A Guthrie - Speaking and Teaching: Siemens, Bayer Healthcare
The following people have nothing to disclose: Eleanor Taylor, Rebecca Jones,
Background and aims: Potent nucleos(t)ide analogs (NA) reduce the risk of hepatocellular carcinoma (HCC) in chronic hepatitis B (CHB) patients, but significant numbers of patients still develop HCC during maintenance NA therapy Therefore,
it is clinically imperative to identify high risk patients whose serum HBV DNA levels are adequately controlled by NAs Liver volume decreases in the early phase of cirrhosis before clinically evident biochemical and hematological changes The aim of this study was to assess the significance of liver volume
in the prediction of HCC development in CHB on entecavir
therapy Methods: A retrospective cohort of 170 patients was
constructed who started entecavir for more than 12 months as
an initial antiviral therapy for chronic hepatitis B Liver volume was measured from the portal phase images of multidetector helical CT which was taken during surveillance for HCC Ver-satile wand tool of ImageJ software was utilized for correct recognition of edges of liver Potential predictors of HCC were
evaluated by Cox proportional hazard model Results: The
cumulative incidence of HCC was 3 6%, 4 8% and 16 4% at
1, 3 and 5 years, respectively Compared to controls, patients who developed HCC during entecavir therapy had lower base-
line liver volumes (1038 ± 307 ml vs 1236 ± 329 ml; p =
0 002) Correction of body surface area, liver volumes divided
by calculated body surface area, still showed significant
differ-ences (601 ± 160 ml vs 702 ± 163 ml, p = 0 002)
Univari-ate analysis showed that low plUnivari-atelet counts and liver volume were significant predictors of HCC whereas low albumin or prolonged prothrombin time were not significant Multivariate analysis confirmed that liver volume < 700 mL independently
increased the risk of HCC (HR: 6 69, CI: 1 93–23 07, p =
0 003) for HCC, along with platelet count < 120K/mm3 (HR:
4 67, CI: 2 03-10 77, p < 0 001) Conclusions: Liver volume is
an independent predictor of HCC in CHB on entecavir therapy Disclosures:
The following people have nothing to disclose: Chung Seop Lee, BeomHee Kim, Jung Wha Chung, Ju Hyun Lee, Sanghyuk Im, Eun Sun Jang, Sook-Hyang Jeong, Jin-Wook Kim
Trang 38Translating the ABC-02 trial into daily practice:
Out-come of palliative treatment in patients with advanced
biliary tract cancer treated with gemcitabine and
cispla-tin
Joeri Dierks 2 , Marcia Gaspersz 1 , Ali Belkouz 2 , Jeroen van Vugt 1 ,
Robert J Coelen 3 , Ferry Eskens 4 , Jan-Willem de Groot 5 , bert-Jan
ten Tije 6 , Wim Meijer 7 , Hans Pruijt 8 , Theo van Voorthuizen 9 ,
Dick Johan van Spronsen 10 , Marjolein Rentinck 11 , Diederik ten
Oever 12 , Jitty Smit 13 , Hans Martin Otten 14 , Thomas van Gulik 3 ,
Hanneke Wilmink 2 , Bas Groot Koerkamp 1 , Heinz-Josef Klümpen 2 ;
1 Surgery, Erasmus University Medical Center, Rotterdam,
Nether-lands; 2 Medical Oncology, Academic Medical Center, Rotterdam,
Netherlands; 3 Surgery, Academic Medical Center, Amsterdam,
Netherlands; 4 Medical Oncology, Erasmus University
Medi-cal Center, Rotterdam, Netherlands; 5 Medical Oncology, Isala
Klinieken, Zwolle, Netherlands; 6 Medical Oncology, Amphia
Ziekenhuis, Breda, Netherlands; 7 Medical Oncology, Westfries
Gasthuis, Hoorn, Netherlands; 8 Medical Oncology, Jeroen Bosch
Ziekenhuis, Den Bosch, Netherlands; 9 Medical Oncology, Rijnstate
Ziekenhuis, Arnhem, Netherlands; 10 Medical Oncology,
Canis-ius Wilhelmina Ziekenhuis, Nijmegen, Netherlands; 11 Medical
Oncology, Tergooi Ziekenhuis, Hilversum, Netherlands; 12 Medical
Oncology, Noordwest Ziekenhuisgroep, Alkmaar, Netherlands;
13 Medical Oncology, Gelre Ziekenhuis, Apeldoorn, Netherlands;
14 Medical Oncology, Medical Center Slotervaart, Amsterdam,
Netherlands
Background: Biliary tract cancer (BTC) is an uncommon
can-cer with an unfavorable prognosis Since 2010, the standard
of care for patients with advanced BTC is a combination of
gemcitabine plus cisplatin, based on the landmark Phase III
ABC-02 trial The eligibility criteria in the ABC-02 trial were
e g an ECOG performance status of 0, 1 or 2, an estimated
life expectancy of at least 3 months, and adequate
hemato-logic and biochemical function Our study aims to evaluate
the efficacy and safety of gemcitabine and cisplatin in patients
with advanced cholangiocarcinoma and gallbladder cancer in
daily practice that meet the criteria for the ABC-02 trial in
com-parison to patients who did not Methods: Patients diagnosed
with advanced BTC between 2010 and 2015 with an
indica-tion for gemcitabine and cisplatin were included We divided
these patients into 3 groups: (I) patients who received
chemo-therapy and met the criteria of the ABC-02 trial and (II) patients
who received chemotherapy and did not meet these criteria
and (III) patients who had an indication for chemotherapy, but
received best supportive care without chemotherapy Primary
outcome was overall survival (OS) and secondary outcome
was progression free survival (PFS) Results: We collected data
of 208 patients Median OS of the 71 patients in group I,
61 patients in group II and 65 patients in group III were 9.7
months (95%CI: 7.1-12.3), 9.4 months (95%CI: 7.2- 11.6)
and 7.6 months (95%CI: 5.0-10.2) (p-value group 1 versus
2 = 0 729, and p-value group 1 and 2 versus 3 = 0 011),
respectively Median PFS was 6.0 months (95%CI: 4.6- 7.4
months) in group I and 4.6 months (95%CI: 3.3- 5.9 months)
in group II (p=0 789) Toxicity and number of dose reductions
were comparable between the two chemotherapy groups
Con-clusion: First line gemcitabine and cisplatin is an effective and
safe treatment for patients with advanced BTC who did not
meet the eligibility criteria for the ABC-02 trial Median OS,
PFS and treatment side effects were comparable between the
two groups
Disclosures:
Heinz-Josef Klümpen - Advisory Committees or Review Panels: IPSEN; Grant/
Research Support: BAYER
The following people have nothing to disclose: Joeri Dierks, Marcia Gaspersz, Ali Belkouz, Jeroen van Vugt, Robert J Coelen, Ferry Eskens, Jan-Willem de Groot, bert-Jan ten Tije, Wim Meijer, Hans Pruijt, Theo van Voorthuizen, Dick Johan van Spronsen, Marjolein Rentinck, Diederik ten Oever, Jitty Smit, Hans Martin Otten, Thomas van Gulik, Hanneke Wilmink, Bas Groot Koerkamp
1269
The efficacy and safety of doxorubicin-eluting bead transarterial chemoembolization over sorafenib for unresectable hepatocellular carcinoma with portal vein invasion
Hyeki Cho 1 , Yuri Cho 1 , Su Jong Yu 1 , Dong Hyeon Lee 1 , Eun Ju Cho 1 , Jeong-Hoon Lee 1 , Hyo-Cheol Kim 2 , Jeong Min Lee 2 , Jin Wook Chung 2 , Joon Koo Han 2 , Yoon Jun Kim 1 , Jung-Hwan Yoon 1 ;
1 Department of Internal Medicine and Liver Research Institute, Seoul National University College of Medicine, Seoul, Korea (the Republic of); 2 Department of Radiology, Seoul National University College of Medicine, Seoul, Korea (the Republic of)
Background and aim: Chemoembolization with doxorubicin drug eluting beads (DEB) is one of treatment modality for advanced hepatocellular carcinoma (HCC) with portal vein invasion (PVI), while conventional transarterial chemoemboli-zation (TACE) is contraindicated for those patients However, currently sorafenib is the only recommended therapy in many guidelines for those patients Therefore, this study aimed to eval-uate the efficacy and safety of DEB-TACE as compared to that
of sorafenib Methods: A total of 301 consecutive HCC patients
with PVI who received DEB-TACE or sorafenib as a sole therapy between 2007 and 2014 were studied, retrospectively DEB-TACE cycles were repeated every 6–8 weeks Primary objec-tive was to compare the OS, in addition to time-to-progression (TTP) and treatment-related adverse events Statistical analysis was performed using Kaplan–Meier method with log-rank test
to compare OS or TTP Results: A total of 48 patients with PVI
who were treated with DEB-TACE as a sole and first therapy (DEB-TACE group) and 253 patients who received sorafenib (sorafenib group) were included There was no significant dif-ference in the level of PVI among the two groups DEB-TACE
group showed superior OS (P = 0 024) and TTP (P = 0 045)
as compared to sorafenib group The TTP was significantly longer in the DEB-TACE group than in the sorafenib group
(12 2 and 3 3 months, respectively, P = 0 009) Subgroup
analysis showed that even among the patients with main portal vein invasion, DEB-TACE group (n = 13) experienced longer
TTP (P = 0 048) as compared to sorafenib group (n = 104)
DEB-TACE group experienced significantly lower risk of
treat-ment-related adverse event compared to sorafenib group (P <
0 05) Conclusions: In our study, DEB-TACE showed both OS
and TTP advantages over sorafenib for advanced HCC patients with PVI with low complication rate
Disclosures:
Jeong Min Lee - Advisory Committees or Review Panels: Bayer healthcare Yoon Jun Kim - Grant/Research Support: Bristol-Myers Squibb, Roche, JW Cre- agene, Bukwang Pharmaceuticals, Handok Pharmaceuticals, Hanmi Pharma- ceuticals, Yuhan Pharmaceuticals; Speaking and Teaching: Bayer HealthCare Pharmaceuticals, Gilead Science, MSD Korea, Yuhan Pharmaceuticals, Samil Pharmaceuticals, CJ Pharmaceuticals, Bukwang Pharmaceuticals, Handok Phar- maceuticals
The following people have nothing to disclose: Hyeki Cho, Yuri Cho, Su Jong Yu, Dong Hyeon Lee, Eun Ju Cho, Jeong-Hoon Lee, Hyo-Cheol Kim, Jin Wook Chung, Joon Koo Han, Jung-Hwan Yoon
Trang 39♦ Denotes AASLD Presidential Poster of Distinction
1270
Hepatitis C virus replication promotes p53 degradation
through chaperone-mediated autophagy (CMA)
Yucel Aydin 1 , Srinivas Chava 2 , Animesh Chatterjee 2 , Partha K
Chandra 2 , Luis A Balart 1 , Hua Lu 3 , Srikanta Dash 2,1 ; 1
Gastroen-terology, Tulane University Health Sciences Center, New Orleans,
LA; 2 Pathology and Lab Medicine, Tulane University Health
Sci-ences Center, New Orleans, LA; 3 Biochemistry, Tulane University
Health Sciences Center, New Orleans, LA
Introduction: Chronic hepatitis C virus (HCV) infection is the
leading cause of liver cirrhosis and hepatocellular carcinoma
(HCC) worldwide However, the mechanism(s) for how the
long-lasting HCV infection leads to HCC development is unclear
Aim: We examined the stability of wild type and mutant p53
using HCV infected primary human hepatocytes culture and
persistently infected Huh-7 5 cells Method: Primary human
hepatocytes and Huh 7 5 were infected with HCV pJFH-ΔV3
Rluc (genotype 2a virus) Replication of HCV was confirmed by
the measurement of the Renilla luciferase activity and
expres-sion of viral proteins by Western blot and immunostaining The
unfolded protein response (UPR) associated with ER stress, as
well as induction of macroautophagy and chaperon-mediated
autophagy (CMA) in the infected culture was measured in a
kinetic study by Western blot analysis and immunostaining The
effect of ER-stress and autophagy on the stability of p53
deg-radation by MDM2 and CMA was investigated Results: We
found that persistent HCV infection of primary human
hepato-cytes and Huh-7 5 cells leads to chronic ER-stress and
activa-tion of unfolded is protein response (UPR) genes The ER-stress
response due to HCV activated macroautophagy and
chap-eron-mediated autophagy (CMA) in infected primary human
hepatocytes as well as Huh-7 5 cells We found that persistent
HCV replication induces degradation of wild-type p53 and its
target gene (p21) in infected primary human hepatocytes
Per-sistent HCV replication in Huh-7 5 cells promoted degradation
of mutant p53 and p21 through induction of MDM2 Inhibition
of p53/MDM2 interactions by small molecule (Nutlin-3) did
not rescue the p53 degradation, indicating that HCV infection
induced degradation of p53 independent of MDM2 pathway
Interestingly, our results showed that HCV degrades both wild
type and mutant p53 through CMA in a lysosome-dependent
mechanism Inhibition of CMA by silencing LAMP2A restored
p53 degradation We also verified that HSC70 and LAMP2A
levels are high and p53 levels are decreased significantly in
HCV-infected cirrhotic livers due to CMA induction, compared
to the cirrhotic liver associated with non-viral etiology
Fur-thermore, we showed that interferon-lambda based antiviral
treatment inhibit HCV replication and restore p53 levels
com-pared to untreated culture Conclusion: Our results indicate that
HCV promotes degradation of tumor suppressor p53 as a
cel-lular protective mechanism to survive under ER-stress We
pro-pose that the long-term suppression of p53 activity increases
genomic instability and development of HCC
Disclosures:
Luis A Balart - Advisory Committees or Review Panels: abbvie; Grant/Research
Support: Merck, Gilead Sciences, Bristol Myers Squibb, abbvie, Merck, Gilead
Sciences, Bristol Myers Squibb, Eisai, takeda, GI Dynamics, tobira; Speaking
and Teaching: Merck, Merck, Merck, Merck, Abbvie, janssen
The following people have nothing to disclose: Yucel Aydin, Srinivas Chava,
Animesh Chatterjee, Partha K Chandra, Hua Lu, Srikanta Dash
1271
Among Adults with Hepatocellular Carcinoma in the United States, Low Income Patients Have Significantly Lower Overall Survival Independent of Tumor Stage and Treatment Received
Melissa Yan 2 , John Ha 2 , Aristeo Lopez 3 , Jennifer Wang 4 , Benny Liu 1 , Taft Bhuket 1 , Robert J Wong 1 ; 1 Gastroenterology and Hepa- tology, Alameda Health System - Highland Hospital, Oakland, CA;
2 Medicine, University of Texas, Houston, TX; 3 Medicine, Alameda Health System - Highland Hospital, Oakland, CA; 4 Medicine, Cal- ifornia Pacific Medical Center, San Francisco, CA
Background: While early diagnosis and treatment via effective screening and surveillance programs improves hepatocellular carcinoma (HCC) outcomes, it remains unclear whether socio-economic disparities as measured by household income may affect timely access to care, receipt of treatment, and overall
survival among adults with newly diagnosed HCC Purpose:
To evaluate household income-specific disparities on overall HCC survival among adults with newly diagnosed HCC in the
U S Methods: Using data from the 2003-2013 Surveillance,
Epidemiology, and End Results database, a population-based cancer registry in the U S , we performed a retrospective cohort study to examine household income specific disparities in over-all survival among adults with HCC Using county-level data, household income was grouped into quartiles based on pro-portion of patients with incomes less than 150% of the federal poverty line Survival differences by household income quar-tiles were evaluated by Kaplan Meier methods and multivariate Cox proportional hazards models, with the final multivariate model including adjustments for age, sex, race/ethnicity, HCC
tumor stage, HCC treatment, and year of diagnosis Results:
Among the 61,594 adults with HCC during the study period,
24 3% were in the highest income quartile, whereas 26 7% were in the lowest household income quartile Overall 1-year survival was significantly higher among individuals in the wealthiest income quartile compared to those in the poorest income quartile (44 9% vs 39 1%, p<0 01) Similar trends were observed even after adjusting for tumor stage (localized HCC, 1-year survival: 59 8% (wealthiest quartile) vs 54 2% (poorest quartile), p<0 01) On multivariate regression, com-pared to HCC patients in the wealthiest income quartile, those from the poorest income quartile had significantly higher over-all HCC mortality (HR, 1 10; 95% CI 1 01-1 20, p=0 037)
(Table) Conclusion: Among U S adults with newly diagnosed
HCC, low-income patients had significantly higher mortality compared to high-income patients even after correcting for HCC tumor stage and HCC treatment received, highlighting the significant impact of income inequality on HCC outcomes Multivariate Cox Proportional Hazards Model for Overall HCC Survival
Model adjusted for age, sex, race/ethnicity, HCC tumor stage, HCC treatment, and year of diagnosis
Trang 40Biological significance of a disintegrin and
metallopro-teinase 21 expression in hepatocellular carcinoma
Masaaki Takamura 1 , Hiroki Honda 1 , Satoshi Yamagiwa 1 ,
Naruhiro Kimura 1 , Toru Setsu 1 , Kentaro Tominaga 1 , Hiroteru
Kamimura 1 , Takuya Genda 2 , Yasunobu Matsuda 3 , Toshifumi
Wakai 4 , Shuji Terai 1 ; 1 Gastroenterology and Hepatology, Niigata
University Graduate School of Medical and Dental Sciences,
Niigata, Japan; 2 Gastroenterology and Hepatology, Jntendo
Uni-versity Shizuoka Hospital, Izunokuni, Japan; 3 Medical Technology,
Niigata University Graduate School of Health Sciences, Niigata,
Japan; 4 Digestive and General Surgery, Niigata University
Gradu-ate School of Medical and Dental Sciences, Niigata, Japan
[Background and aim] We have previously reported that
hepa-tocellular carcinoma cell lines with high motility in vitro have
a high rate of intrahepatic metastasis in vivo This indicates
that cell motility plays an important role in the malignancy of
hepatocellular carcinoma Here we investigated the biological
significance of a disintegrin and metalloproteinases (ADAMs),
multi-functional molecules involved in cell motility, in
hepato-cellular carcinoma [Method] Expression of 12 ADAM
fam-ily members with putative metalloproteinase activity has been
confirmed by real time PCR in the highly motile and highly
metastatic hepatocellular carcinoma cell line KYN-2 as well as
in PLC/PRF/5 and HepG2, which show low motility and low
metastasis ADAM21, which is highly expressed in KYN-2, has
garnered considerable research attention Therefore, we
estab-lished a KYN-2 cell line stably expressing ADAM21 siRNA
and investigated its invasion, migration, and proliferation We
also generated an anti-ADAM21 mouse monoclonal antibody
and performed immunostaining of 96 cases of International
Union Against Cancer with tumor, node and metastasis
classi-fication stage I/II (initial onset with resection of hepatocellular
carcinoma) Furthermore, we investigated ADAM21 expression
and other clinicopathological factors [Results] ADAM17, 21,
and 28 were highly expressed in KYN-2 cells The KYN-2 cell
line stably expressing ADAM21 siRNA showed significantly
lower capacity for invasion (p<0 001), migration (p<0 001),
and proliferation (p=0 003) than the control Among the cases
where the hepatocellular carcinoma had been resected, 16
patients (16 7%) were ADAM21 positive, and there were
sig-nificantly more cases positive for vascular invasion (p=0 025)
Event-free survival rate (p=0 002) and overall survival rate
(p=0 002) were both significantly lower in cases that were
positive for ADAM21 than in the negative cases Multivariate
analysis indicated that tumor multiplicity (RR 2 283, p=0 003)
and ADAM21 positivity (RR 3 296, p<0 001) were
indepen-dent risk factors for recurrence and that preoperative serum
alpha-fetoprotein level (>20 ng/mL) (RR 2 598, p=0 010) and
ADAM21 positivity (RR 2 961, p=0 008) were independent
prognostic factors [Conclusion] ADAM21 may be involved
in the malignancy of hepatocellular carcinoma and could be
a novel prognostic and recurrence-predictive marker for this
disease
Disclosures:
The following people have nothing to disclose: Masaaki Takamura, Hiroki
Honda, Satoshi Yamagiwa, Naruhiro Kimura, Toru Setsu, Kentaro Tominaga,
Hiroteru Kamimura, Takuya Genda, Yasunobu Matsuda, Toshifumi Wakai, Shuji
Terai
1273
Predictors of Inadequate Ultrasound Quality for cellular Carcinoma Surveillance in Patients with Cirrho- sis
Hepato-Okeefe L Simmons 1 , David T Fetzer 2 , Takeshi Yokoo 2 , Jorge A Marrero 1 , Adam C Yopp 3 , Travis Browning 2 , Amit G Singal 1 ;
1 Internal Medicine, University of Texas Southwestern, Irving, TX;
2 Radiology, UT Southwestern Medical Center, Dallas, TX; 3 gery, UT Southwestern Medical Center, Dallas, TX
Sur-Background: Abdominal ultrasound fails to detect over fourth of hepatocellular carcinoma (HCC) at an early stage in patients with cirrhosis Identifying patients in whom ultrasound
one-is of inadequate quality can inform interventions to improve
surveillance effectiveness Aims: Evaluate ultrasound quality
and identify clinical predictors of inadequate ultrasound ity among a large cohort of patients with cirrhosis undergoing
qual-HCC surveillance Methods: We performed a retrospective
cohort study among patients who underwent ultrasound ination for a cirrhosis-related indication between April 2015 and October 2015 Three fellowship-trained abdominal radiol-ogists collectively reviewed all ultrasound exams and catego-rized exam quality as definitely adequate, likely adequate, likely inadequate, and definitely inadequate to exclude liver lesions Quality assessment was based on impression of overall exam quality based on combination of anatomical coverage, visual clarity of the liver, penetration, and any other exam limitations We performed multivariable logistic regression to determine characteristics associated with inadequate ultra-
exam-sound quality Results: We identified 941 patients; mean age
was 57 years, and 64% were men The most common liver ease etiologies were 48% hepatitis C, 18% alcohol, and 12% NASH Most patients had compensated cirrhosis, with 69% Child Pugh A cirrhosis Ultrasound quality was inadequate for exclusion of liver masses in 191 (20 3%) patients - 134 defi-nitely inadequate and 57 likely inadequate The most common reasons for inadequate ultrasound exams were rib shadowing and inadequate beam penetration allowing visualization of less than two-thirds of the hepatic parenchyma There were 33 patients with rib shadowing, 26 with poor beam penetration, and 82 with both quality issues In multivariable analysis, inad-equate quality was associated with male gender (OR 1 68, 95%CI 1 14-2 48), body mass index category (OR 1 67, 95%CI 1 45-1 93), Child Pugh B or C cirrhosis (OR 1 93, 95%CI 1 32-2 81), alcohol-related cirrhosis (OR 2 11, 95%CI
dis-1 33-3 37), NASH cirrhosis (OR 2 87, 95%CI dis-1 7dis-1-4 80), and inpatient status (OR 1 55, 95%CI 1 01-2 37) Ultrasounds were inadequate in over one-third of patients with Child Pugh
C cirrhosis, BMI >35, or NASH cirrhosis Conclusions: One
in 5 ultrasounds in patients with cirrhosis are inadequate for exclusion of HCC, which can contribute to surveillance fail-ure Alternative surveillance modalities may be necessary in subgroups prone to inadequate ultrasounds including obese patients, those with Child Pugh B or C cirrhosis, and those with alcohol- or NASH-related cirrhosis