30 Phase II Clinical Trial of Gene Therapy for Adenosine Deaminase Deficient Severe Combined Immune Deficiency (ADA SCID) Using a γ Retroviral Vector Molecular Therapy Volume 23, Supplement 1, May[.]
Trang 1Molecular Therapy Volume 23, Supplement 1, May 2015
Hematologic and immunologic diseases: clinical trials and observations
28 Intravenous Administration of Lentiviral
Vectors Expressing Hyperactive Factor IX
Converts Severe Into Mild Hemophilia B in a
Canine Model
Alessio Cantore,1 Michela Milani,1 Tongyao Liu,2 Patrizia Della
Valle,3 Armando D’Angelo,3 Thierry VandenDriessche,4 Marinee
Chuah,4 Haiyan Jiang,2 Timothy Nichols,5 Luigi Naldini.1,6
1 San Raffaele Telethon Institute for Gene Therapy, Milan, Italy;
2 Biogen Idec, Cambridge; 3 Coagulation Service, San Raffaele
Scientific Institute, Milan, Italy; 4 Free University of Brussels,
Brussels, Belgium; 5 University of North Carolina, Chapel Hill;
6 Vita Salute San Raffaele University, Milan, Italy.
Lentiviral vectors (LVs) are attractive vehicles for liver-directed
gene therapy by virtue of their ability to stably integrate in the
genome of target cells and the lack of pre-existing immunity
against vector components in most humans Over the past years,
we have developed a LV platform that can achieve stable transgene
expression in the liver, induce transgene-specific immune tolerance
and establish correction of hemophilia in mouse models upon
systemic administration This LV is designed to stringently target
transgene expression to hepatocytes through transcriptional and
microRNA-mediated regulation We then investigated the efficacy
and safety profile of portal vein administration of LVs expressing
wild-type, codon-optimized (c.o.) or c.o and hyperactive factor IX
(FIX) in a canine model of hemophilia B We produced large-scale
batches of LVs qualified for in vivo administration and treated adult
hemophilia B dog by portal vein administration We observed
long-term stable reconstitution of canine FIX activity up to 1% of normal
and significant amelioration of the clinical phenotype in 3 treated dogs
(>9 years cumulative follow up) LV infusion was associated with
transient signs of inflammation and mild hepatotoxicity, which could
be abrogated by pretreatment with anti-inflammatory drugs There was
no detectable long-term toxicity or development of FIX inhibitors
In the perspective of clinical translation and to increase therapeutic
efficacy, we next treated an 11-kg, hemophilia B dog by peripheral
vein administration of LVs expressing the c.o and hyperactive canine
FIX at a 5-fold higher dose than those previously administered At the
current follow-up (3 months after gene therapy) FIX activity is 6-9%
of normal Intravenous LV administration, coupled with a 1-day
anti-inflammatory and anti-histamine pre-treatment, induced mild and
self-limiting leukopenia and elevation of aminotransferases Treatment
of more hemophilia B dogs is underway to confirm and extend these
results Overall, our studies, which suggest comparable efficacy of
LV by both portal and peripheral vein administration, position
LV-mediated liver gene therapy for further pre-clinical development and
clinical translation LVs may thus complement other available vectors
to address some of the outstanding challenges posed by liver gene
therapy of hemophilia and conceivably other diseases
29 Clinical Trial Showing EPO-Independence
for 7 Months by Prolonged Secretion of
Autologous EPO by TARGT™
Shany Blum,1 Nir Shapir,1 Reem Miari,1 Atar Liran,1 Benny
Lerner,1 Keren Doenyas-Barak,2,3 Shai Efrati,2,3 Doron Schwartz,4
Gil Chernin,4 Garry A Neil.5
1 Medgenics Medical Israel, Ltd., Misgav, Israel; 2 Research &
Development Unit and Nephrology Unit, Assaf-Harofeh Medical
Center, Zerifin, Israel; 3 Sackler Scholl of Medicine, Tel Aviv
University, Tel Aviv, Israel; 4 Nephrology and Dialysis Department,
The Tel Aviv Sourasky Medical Center, Tel Aviv, Israel;
5 Medgenics, Inc., Wayne, PA.
Recombinant human EPO (rHuEPO) along with iron
supplementation corrects anemia in most patients with End Stage
Renal Disease (ESRD) but is associated with supra-physiological peak serum concentration (Cmax) of EPO and may cause thromboembolic complications
The Transduced Autologous Restorative Gene Therapy system
(TARGT™) is an ex-vivo gene therapy that provides autologous,
continuous proteins or peptide therapy in the physiological range The system consists of several 2 x 30 mm pieces of dermal tissue (Micro-Organ, MO), extracted under local anesthetic in which its fibroblasts cells are transduced with a Helper-Dependent Adenoviral Vector (HDAd) containing the EPO gene expression cassette After culture, and measurement of EPO production, one or more transduced MOs (TARGTs) are re-implanted as required for delivering the target dose Patients are treated with local steroid injection to stabilize secretion The system allows dose flexibility and the TARGTs may be removed
or added according to the in-vivo secretion levels
We present here initial results from an-ongoing open label ascending dose clinical study of TARGTEPO in patients with anemia due to ESRD We have completed the enrollment of patients in the first out of 3 cohorts (the low dose) with 6 EPO-dependent patients treated with a total of up to 3 TARGTEPO units each, secreting a total
of 25 IU/Kg/day of autologous EPO All patients continued their previous regimen of intravenous supplemental iron
Patients follow-up post implantation is still ongoing with one patient being followed with stable EPO secretion and resulting stable
Hb for over 7 months Results obtained suggest stabilization of serum EPO levels at the physiological range of £20 mIU/ml and the resulting
Hb levels between 9-12 g/dL without rHuEPO or transfusion while TARGTs are still functioning Comparative analysis of serum EPO levels revealed significantly lower Cmax with TARGTEPO compared
to rHuEPO Also, comparison of extrapolated Area Under the Curve
(AUC) of rhEPO vs actual TARGTEPO AUC, revealed that TARGTEPO maintained Hb within the desired range in patients with an order of magnitude smaller exposure to EPO compared to rHuEPO This observation may have significant clinical benefits No treatment related serious adverse events have been reported TARGTEPO is a promising novel therapy for anemia and potentially for other protein deficient diseases
30 Phase II Clinical Trial of Gene Therapy for Adenosine Deaminase-Deficient Severe Combined Immune Deficiency (ADA-SCID) Using a γ-Retroviral Vector
Kit L Shaw,1 Elizabeth Garabedian,2 Rob Sokolic,2 Provaboti Barman,1 Alejandra Davila,1 Christopher Silvin,2 Satiro de Oliveira,1 Ami J Shah,1 Dayna Terrazas,1 Denise Carbonaro,1 Sabine Geiger,1 Suparna Mishra,1 Aaron Cooper,1 Monika Smogorzewska,3 Jayashree Jagadeesh,2 Michael S Hershfield,4 Alan Wayne,2 Gay M Crooks,1 Theodore Moore,1 Fabio Candotti,2 Donald B Kohn.1
1 University of California Los Angeles, Los Angeles, CA; 2 Mark
O Hatfield Clinical Research Center, Bethesda, MD; 3 Children’s Hospital Los Angeles, Los Angeles, CA; 4 Duke University, Durham, NC.
We report follow-up of subjects treated in a Phase II study of gene therapy for ADA-SCID Between 2009 and 2012, ten ADA-deficient SCID patients were treated by γ-retroviral-mediated gene transfer (MND-ADA) to their bone marrow CD34+ cells The subjects were given non-myeloablative chemotherapy (busulfan @ 90 mg/m2) and were withdrawn from PEG-ADA enzyme replacement therapy (ERT) prior to infusion of autologous gene-modified cells Subject age at the time of treatment ranged from 3 months to 15 years (median = 11.5 months) Follow-up times range from 2 to 5 years All but one subject, who was 15-years old at the time of treatment, remain off PEG-ADA ERT with immune reconstitution that reached maximal level between
Trang 2Molecular Therapy Volume 23, Supplement 1, May 2015 Copyright © The American Society of Gene & Cell Therapy
S14
Stem Cell engineering and therapieS i
6 and 12 months after transplant and was maintained thereafter
Vector marking in peripheral blood cells remained consistently
detectable (> 0.1 copy/PBMC and ≥ 0.003 copy/granulocyte) at 2
years and later after transplant in subjects who discontinued ERT
These subjects also had PBMC ADA enzymatic activity in the
normal range and red blood cell deoxynucleotide levels below 10%
Three subjects have discontinued intravenous immunoglobulin; five
subjects have discontinued prophylactic antibiotics All subjects
have polyclonal gene marking with no sign of lymphoproliferative
disease The subjects remain in good health without infections or
other complications
Stem Cell Engineering and Therapies I
31 EncorStat®: A Human Donor Cornea
Genetically Engineered To Resist Rejection in High
Risk Patients
Laura McCloskey,1 Maria Parker,2 Vicky Kennedy,1 Trevor
McFarland,2 Matt Hartzell,2 Binoy Appukuttan,3 Tim Stout,4 Khilan
Shah,5 Frank Larkin,5 Simon Chandler,1 Kyriacos Mitrophanous,1
Scott Ellis.1
1 Oxford BioMedica (UK) Ltd, Oxford, United Kingdom; 2 Oregon
Health & Sciences University, Portland, OR; 3 Flinders University,
Adelaide, Australia; 4 Baylor College of Medicine, Houston; 5 NIHR
Moorfields Biomedical Research Centre, Moorfields Eye Hospital,
London, United Kingdom.
Corneal transplantation is one of the most successful transplant
procedures because of the relatively immune-privileged status of
the eye and the avascularity of the cornea However, normal corneal
immune privilege can be eroded by neovascularization by providing
a route of entry for immune-mediating cells, leading to subsequent
irreversible immunological rejection of the corneal graft, the most
common reason for graft failure In high risk patients, which account
for >20% of the 100,000 transplants carried out worldwide each year,
the rejection rate can be very high (50-90%), particularly if there is
pre-existing vascularization of the recipient corneal bed In these
patients the prognosis is extremely poor, with grafts failing at an
accelerating rate to the point where patients are no longer considered
suitable for further transplants and are left blind, despite an otherwise
healthy eye It is therefore not surprising that neovascularization
(both pre- and post-grafting) is a significant risk factor for corneal
graft failure Neovascularization is thus an attractive target to prevent
corneal graft failure due to rejection
EncorStat® is a human donor cornea modified prior to transplant
by the ex vivo delivery of the genes encoding secretable forms
of the angiostatic human proteins, endostatin and angiostatin, by
a lentiviral vector, derived from the Equine Infectious Anaemia
Virus (EIAV), which prevents subsequent rejection by suppressing
neovascularization
Modified rabbit corneas have been evaluated in two different
models of corneal graft rejection, a highly aggressive model in which
rejection is driven by the retention of thick graft sutures, and a less
aggressive model in which rejection is driven by pre-vascularizing
the recipient corneal bed prior to surgery In this latter model thin
sutures are used to secure the graft that are removed two weeks
following surgery, which is more analogous to the clinical setting
The process to generate EncorStat® corneas has been optimized to
secrete substantial and persistent levels of angiostatic proteins with
very little shedding of residual vector These corneas substantially
suppress corneal neovascularization, opacity and subsequent rejection
in both rabbit models of cornea graft rejection
The non-clinical data to be presented support the evaluation of
EncorStat® corneas in a First-in-Man trial With support from the UK
Technology Strategy Board (Innovate UK), this trial will be conducted
in 2016, following completion of non-clinical safety studies and GMP vector manufacture this year An outline of this clinical trial design will also be presented
32 Safety Studies Towards a Combined Cell and Gene Therapy To Treat Critical Limb Ischemia
Julie R Beegle,1 Stefanos Kalomoiris,1 Nataly L Magner,1 Jan A Nolta,1 Fernando A Fierro.1
1 University of California, Davis, Davis, CA.
Over 240 clinical trials are in progress worldwide using mesenchymal stem cells (MSCs) Some critical advantages of MSCs include simple isolation and expansion, and ease for allogeneic applications due to their immune evading properties MSCs produce many paracrine signals that promote tissue regeneration, though
at sub-therapeutic levels We have therefore developed a product that combines cell and gene therapy, by engineering MSCs to over-express vascular endothelial growth factor (VEGF) Here we show our preclinical data that not only confirms the efficacy of our product (MSC-VEGF), but addresses multiple safety aspects Here, MSCs are transduced with a GMP-grade lentiviral vector to obtain an average
of one copy per cell (ranging from 0.8 to 1.3), minimizing occurrence
of insertional mutagenesis The secretion of VEGF is optimized
by using a previously clinically evaluated constitutive promoter (MNDU3) and an enhancer element acting in cis (WPRE) Safety studies at this level of secretion (approx 80pg/ml VEGF secreted per 1000 cells per 24 hours) did not cause edema or hemangiomas
in immune deficient NOD/SCID IL-2R-gnull/null (NSG) mice The genomic stability of MSC-VEGF was addressed by karyotyping analysis, where no abnormalities were found in MSCs transduced with increasing viral loads, and by PCR to evaluate potential genomic rearrangement of the insert To rule out tumorigenicity of MSC-VEGF, cells transduced with increased viral load were in injected into NSG mice In contrast to 3 distinct positive controls used, no tumors were detected in over 50 mice treated with MSC-VEGF up to 6 months after injection Additional safety studies included confirmation by flow cytometry of homogeneity of the transduction level of MSC-VEGF, absence of replication competent lentivirus, freedom of endotoxin, mycoplasma and absence VSV-G viral envelope coding plasmid Luciferase-expressing MSC-VEGF consistently decreased
to undetectable levels by 28 days after injection and PCR confirmed the absence of human DNA 6 months after injection Pilot efficacy studies using MSC-VEGF in an immune deficient mouse model of hind limb ischemia (HLI) have been completed MSC-VEGF were injected IM the day after HLI surgery into the ischemic limbs of NSG mice and blood flow was monitored weekly, demonstrating that MSC-VEGF treatment leads to faster revascularization than control treatment Furthermore, we show that a small percentage of injected MSC-VEGF take a pericyte position on blood vessels While future assays are intended as IND-enabling safety studies, these pre-clinical safety and efficacy studies are directed towards our planned Phase
I clinical trial
33 Ex Vivo Gene Therapy for Lysosomal Storage Disease in the CNS: Patient iPSC-Derived Neural Stem Cells Correct Pathology in the MPS VII Mouse Brain
Tagan A Griffin,1 Hayley C Anderson,1 John H Wolfe.1,2
1 Neurology, Research Institute of the Children’s Hospital of Philadelphia, Philadelphia, PA; 2 W.F Goodman Center for Comparative Medical Genetics, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA.
Neural stem cells (NSCs) derived from induced pluripotent stem cells (iPSCs) may offer an immunologically compatible delivery