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Tiêu đề Blood Disorders in the Elderly - Part 4
Tác giả Oswald Steward, Arati V. Rao, Harvey Jay Cohen
Trường học Reeve-Irvine Research Center, Departments of Anatomy & Neurobiology, Neurobiology & Behavior, Neurosurgery, University of California at Irvine
Chuyên ngành Hematology and Blood Disorders
Thể loại sách tham khảo
Năm xuất bản 2008
Thành phố Irvine
Định dạng
Số trang 50
Dung lượng 1,29 MB

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Nội dung

The International Myeloma Working Group has defi ned MGUS as the presence of a monoclonal protein in patients without the evidence of multiple myeloma, amyloidosis, Waldenström macroglob-

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InXtroduction

Oswald Steward

Reeve-Irvine research center, departments of anatomy & nurobiology, nurobiology & behavior,

and neurosurgery, university of california at irvine, Irvine, CA 92697

Introduction

Monoclonal gammopathy of unknown signifi cance

(MGUS) affects up to 2% of persons aged 50 years

or over, and about 3% of those older than 70 years

The aim of this chapter is to highlight some of the

features of this disease process and its relationship

to aging It will focus on (1) epidemiology, (2)

biol-ogy of MGUS and the role of various cytokines in

the pathophysiology, (3) correlation of the biology

of MGUS with aging, (4) diagnosis and follow-up

of patients with MGUS, and (5) natural history and

predictors of progression in patients with MGUS

Epidemiology

The monoclonal gammopathies are a group of

dis-orders associated with proliferation of a single clone

(monoclonal) of plasma cells They are characterized

by the secretion of an immunologically homogenous

monoclonal protein (M-protein, M-component,

M-spike, or paraprotein) Each M-protein consists

of two heavy (H) polypeptide chains of the same

class and subclass, and two light (L) chains of the

same type The heavy chains are IgG, IgM, IgA, IgD,

and IgE, while the light-chain types are kappa (κ),

and lambda (λ) In contrast, a polyclonal

gammopa-thy is characterized by an increase in one or more

heavy chains, and in both types of light chains, and

is usually associated with an infl ammatory or

reac-tive process [1] The term monoclonal gammopathy

of unknown signifi cance (MGUS) was fi rst coined by

Biological and clinical signifi cance of monoclonal

gammopathy

Arati V Rao, Harvey Jay Cohen

Kyle et al., to replace the term benign monoclonal

gammopathy, which was misleading because, at diagnosis, it is not known if the disease process will remain stable and asymptomatic, or evolve into symptomatic multiple myeloma (MM) [2]

The International Myeloma Working Group has defi ned MGUS as the presence of a monoclonal protein in patients without the evidence of multiple myeloma, amyloidosis, Waldenström macroglob-ulinemia (WM), or any other B-cell lymphoprolif-erative disorder More specifi cally it is defi ned as an M-spike of 3.0 g/dL or trace or no light chains in a 24-hour urine collection, less than 10% plasma cells

in the bone marrow, and no related organ or sue impairment, i.e., no lytic bone lesions, and the absence of anemia, hypercalcemia, and renal insuf-

tis-fi ciency [1] As for most cancers, especially logic malignancies, this condition demonstrates an increased incidence with age and affects up to 2% of persons50 years and about 3% of those older than

hemato-70 years [3] A screening study conducted in Sweden demonstrated MGUS in 0.1–0.2% of persons aged 30–49 years, 1.1–2.0% of persons 50–79 years, and in 5.7% of persons 80–89 years [4] In a cluster of cases

of MM, Kyle et al were able to detect an M-protein

in 15 of 1200 persons 50 years or older (1.25%) [3], and in France, 303 of 17 968 persons 50 years or

older (1.7%) had an M-protein [5] Crawford et al.

have reported that 10% of 111 persons older than

80 years had an M-protein ranging in concentration from 0.2 to 1.8 g/dL [6] This has also been reiterated

by Cohen et al., who found that 3.6% of 816 persons

70 years or older had an M-protein [7] As with

11

Blood Disorders in the Elderly, ed Lodovico Balducci, William Ershler, Giovanni de Gaetano

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MM, the incidence of MGUS is higher in African–

Americans than in whites, and in one study the

prevalence of an M-protein was 8.4% in 916 African–

Americans [7] In contrast, the incidence of MGUS is

only about 2.7% in elderly Japanese patients [8] The

monoclonal protein in MGUS is most commonly

IgG (73%), followed by IgM (14%) and IgA (11%) The

light chains in MGUS most commonly involve the

κ molecules (62%) [9].

MGUS is frequently a single abnormality, but it

may be associated with many other diseases, as

would be expected in the elderly populations Most

common associations have been with the B-cell

lymphoproliferative disorders like chronic

lym-phocytic leukemia, non-Hodgkin lymphoma, and

hairy-cell leukemia [10] One prospective study

showed that MGUS was detected in 1.1% of patients

with solid tumors referred for systemic

chemother-apy [11] A third of patients with chronic neutrophilic

leukemia, which is a rare disorder characterized by

persistent leukocytosis of mature neutrophils, have

an elevated M-protein [12] It has also been seen

in Gaucher’s disease [13], myelofi brosis, hepatitis

C infection, HIV infection [14], rheumatoid

arthri-tis [15], and other related disorders Interestingly,

MGUS has been observed after liver, renal, and

bone-marrow transplantation, and in these patients

the development of an M-protein correlated with

the presence of a viral infection, e.g.,

cytomegalovi-rus infection [16,17]

The gammopathies can be further classifi ed as:

• benign (IgG, IgM, IgA, IgD)

• associated with malignancies that are not known

to produce monoclonal proteins

• idiopathic Bence-Jones proteinuria [18,19]

• biclonal gammopathies [20]

• triclonal gammopathies [21]

Idiopathic Bence-Jones proteinuria is a condition

in which patients excrete large amounts of

mono-clonal light chains (Bence-Jones protein) and follow

a benign course A small series of seven patients

revealed no evidence of malignant plasma-cell

dis-order, and no serum M-protein, but urine light-chain

excretion of 1 g/day [19] In all these patients the

plasma cell labeling index was low After a follow-up

of 7–28 years, three of the seven patients, developed

MM, while two other patients developed matic MM, and evolving MM One patient devel-oped primary amyloidosis after 12 years, and two patients continued to have stable levels of Bence-Jones proteins The authors suggested that patients with idiopathic Bence-Jones proteinuria should be monitored regularly and indefi nitely

asympto-Of patients with a gammopathy of unknown signifi cance, 3–4% have a biclonal gammopathy, characterized by the production of two different M-proteins This may be due to the proliferation of two different clones of plasma cells each producing

an unrelated monoclonal protein, or it may result from a single clone of plasma cells producing two M-proteins In a series of 57 biclonal gammopathy patients, the most common diagnoses were biclonal gammopathy of undetermined signifi cance (65%), multiple myeloma (16%), and lymphoproliferative disease (19%) [20] The clinical features and response

to therapy were similar to patients with monoclonal gammopathy Of note, serum protein electrophore-sis (SPEP) may produce only a single band on the acetate strip, and the biclonal gammopathy may be recognized only by immunofi xation

Triclonal gammopathy has also been reported, and these patients may also have underlying lym-phoproliferative disorder, or a nonhematologic condition causing production of three different immunoglobulins [21]

Biology of MGUS and the role of various cytokines

It is well known that germinal-center B cells uniquely modify the DNA of immunoglobulin (Ig)genes through sequential rounds of somatic hyper-mutation, antigen selection, and IgH switch recom-bination Post-germinal-center plasmablasts can generate plasmablasts that have successfully com-pleted somatic hypermutation and IgH switching before migrating to the bone marrow, where stromal cells enable terminal differentiation into plasma

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140 Arati V Rao, Harvey Jay Cohen

cells MGUS and MM are both characterized by

the accumulation of transformed plasmablasts or

plasma cells in the bone marrow [22,23] However,

MGUS is less proliferative than MM, with 1% cells

synthesizing DNA [24] Gene expression profi ling

data has demonstrated a higher level of cyclin D1,

D2, or D3 mRNA in patients with MGUS and

sub-sequently with MM, when compared to normal

plasma cells [25] This allows the plasma cells to be

more susceptible to proliferative stimuli, with

selec-tive expansion, after interacting with bone-marrow

stromal cells that produce interleukin 6 (IL-6) and

other cytokines There is also some evidence that the

Rb protein which controls the cell-cycle restriction

point from G1 to S phase might be dysregulated due

to methylation of p16, which can inhibit cyclinD/

CDK4 and thus prevent phosphorylation of Rb [26]

This, along with deletion of chromosome 13, may

be the earliest change in MGUS that allows

progres-sion to MM [24] In addition, activating mutations of

N-ras and K-ras are absent with MGUS but are seen

in 30–40% patients with MM [27]

Role of IL-6, IL-6R, IL-1 β, and TNF-α

The function, differentiation, and survival of

hematopoietic cells are governed by the presence

of certain cytokines These cytokines in turn require

expression of an appropriate cellular receptor to

exert their many biologic effects The

develop-ment of MGUS and MM is dependent upon

differ-ent cytokines like granulocyte colony-stimulating

factor (G-CSF), interferon alpha (IFN-α), leukemia

inhibitory factor (LIF), IL-11, tumor necrosis

fac-tor alpha (TNF-α), and IL-6 IL-6, a multifunctional

cytokine, has been thoroughly investigated in MM

and MGUS and may possess the most biologic and

clinical signifi cance [28] The primary function of

IL-6 is to stimulate the differentiation of mature B

cells into plasma cells and also to allow proliferation

of plasmablasts in the bone marrow [29] In addition,

IL-6 is known to inhibit fas- and

dexamethesone-induced plasma-cell apoptosis in vitro [30,31]

Initial clinical observations have suggested that

high serum IL-6 levels correlate with advanced

disease, aggressive disease, and chemotherapy refractoriness [32] Multiple studies have been per-formed to demonstrate that IL-6 stimulates pro-liferation of myeloma cells in vitro, and anti-IL-6 antibodies or IL-6 antisense oligonucleotides can inhibit IL-6-stimulated growth of myeloma cells The autocrine and paracrine functions of IL-6 in myeloma, along with IL-6 transgenic mouse mod-els, has also been studied The expression of IL-6 receptors (IL-6R) by myeloma cells, and responses

in patients treated with anti-IL-6 antibodies, have also been examined in order to study the role of IL-6

in the pathophysiology of MGUS and MM [33,34] More recently, similar fi ndings have also been con-

fi rmed in patients with MGUS Sati et al developed

a dual-color fl uorescence in-situ hybridization

(FISH) technique to investigate the expression of IL-6 mRNA in bone-marrow cells of patients with

MM, with MGUS, and in healthy bone-marrow donors [35] The IL-6 protein could be detected by direct immunofl uorescence in all plasma cells from all patients with MM, and in those with MGUS, with lower levels of expression in patients with MGUS than in those with MM However, neither the IL-6 mRNA nor protein could be detected in normal plasma cells from healthy subjects These data demonstrated that patients with MGUS and MM express the IL-6 mRNA, and support the hypothesis

of autocrine synthesis of IL-6 in these patients Most investigators, however, agree that the contribution

of autocrine IL-6 is minimal, and there are emerging data that the paracrine secretion of IL-6 is the major factor in the pathogenesis of MGUS, MM, and other monoclonal gammopathies [36]

IL-6 production has been detected by Th2 T cells, monocytes, endothelial cells, fi broblasts, and bone-marrow stromal cells, and the latter is probably the major source of IL-6 in monoclonal gammopathies

[37] This has been confi rmed by Klein et al., who

attributed the high production of IL-6 to ent cells of the bone-marrow microenvironment

adher-by demonstrating a spontaneous proliferation of myeloma cells in vitro Recombinant IL-6 was able

to amplify this proliferation, and anti-IL-6 ies were able to inhibit these cells [28]

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antibod-Of note, the signaling of IL-6 is mediated via a

spe-cifi c heterodimer receptor made up of an α chain of

80 kD (IL-6R) and a β transducer chain of 130 kD

(gp130) A remarkable feature of the IL-6 receptor is

the agonist role of its soluble form (sIL-6R), which

is able to bind IL-6 with an affi nity similar to that

of membrane IL-6R Also, the IL-6/sIL-6R complex

is able to bind and activate the gp130 transducer

chain [38,39] Stasi et al investigated the clinical

sig-nifi cance of serum sIL-6R in 81 patients with MGUS,

and 164 patients with MM, and found higher levels

of sIL-6R in the MM patients In a univariate

analy-sis, sIL-6R was a signifi cant but weak prognostic

indicator, and higher levels were associated with

shorter survival [40]

The relationship of IL-6 to other cytokines like

TNF-α and IL-1β has also been well studied TNF-α

and IL-1β are potent inducers of IL-6 production and

play a role in paracrine secretion of IL-6 [41] These

two cytokines, especially IL-1β, are potent osteoclast-

activating factors and play a role in the development

of lytic lesions (Fig 11.1) Lacy et al performed

in-situ hybridization for IL-1β using bone-marrow

aspirates from 51 patients with MM, 7 with

smol-dering myeloma, 21 with MGUS, and 5 healthy

subjects [42] IL-1β mRNA was detected in plasma

cells from a majority of patients with MM (49 of 51) and smoldering myeloma, but only 5 of 21 patients with MGUS, and none of the normal sub-jects had any detectable IL-1β mRNA This contrast

in cytokine expression of IL-6 and IL-1β between

patients with MGUS and MM has also been

demon-strated by Donovan et al [43] TNF- α plays a role in

the production of IL-6 in a dose-dependent fashion

Blade et al mea sured serum levels of IL-6 and TNF- α

in 38 healthy subjects and 100 patients with MGUS IL-6 levels were signifi cantly higher in MGUS than

in healthy controls (p  0.0001) Similarly, TNF-α

levels were signifi cantly higher in MGUS than in

control populations (p 0.015) [44]

More recently, the role of adhesion molecules

in the biology of myeloma has been studied [45] Normal B cells are able to home to certain tissues due to the presence of surface adhesion molecules Myeloma cells may express a variety of surface adhesion molecules such as NCAM (CD56), ICAM (CD54), HCAM (CD44), and others A recent study has also demonstrated impaired osteoblastogenesis

in myeloma, thought to be due to increased levels

of cytokines like IL-1β, TNF-α, and IL-6 which in

MGUS MM-Low LI

IL-6IL-1b

MM-High LI

M-protein

adhesionmolecules

lytic bonelesions

paracrineIL-6

OAFIL-1b

gammopathy of unknown signifi cance; LI, labeling index; OAF, osteoclast-activating factor Adapted from Lacy MQ et al.,

Blood 1999; 93: 300–5 [42].

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142 Arati V Rao, Harvey Jay Cohen

turn led to upregulation of ICAM-1 [46] It has also

been hypothesized that acquisition of NCAM

expres-sion in myeloma is a malignancy-related

phenom-enon NCAM (CD56) is strongly expressed on most

myeloma cells but is not found on normal plasma

cells In one study, CD56 expression in high density

was present in 43 of 57 patients with untreated MM

and in none of 23 patients with MGUS [47] IL-6 has

been shown to increase HCAM (CD44) gene

expres-sion and cause overexpresexpres-sion of all CD44 variant

exons [48] It is thought that these adhesion

mol-ecules play a role in cell-to-cell contact between

myeloma cells and marrow stromal cells, and this

maybe leads to the homing of myeloma cells to the

bone marrow and to the development of osteolytic

bone lesions, and may also play a crucial role in

myeloma cell survival

IL-6 and bone-marrow angiogenesis

There has been a study which demonstrated that

vascular endothelial growth factor (VEGF) expressed

and secreted by myeloma cells stimulates the

expres-sion of IL-6 by microvascular endothelial cells and

the bone-marrow stromal cells In turn, IL-6

stimu-lates the expression of VEGF, which as we know is a

potent stimulator of angiogenesis [49] Numerous

studies have now demonstrated that marrow

angio-genesis parallels tumor progression and correlates

with tumor growth and metastatic potential in

mul-tiple myeloma patients We also have evidence that

bone-marrow angiogenesis progressively increases

along the spectrum of plasma-cell disorders, from

MGUS to advanced myeloma This has been studied

in the bone-marrow samples of 400 patients (76 with

MGUS) by immunohistochemical staining for CD34

to identify microvessels, and compared to normal

bone-marrow samples [50] The median microvessel

density per  400 high-power fi eld was 1.3 in controls,

3 in MGUS, 11 in newly diagnosed MM, and 20 in

relapsed MM Higher-grade angiogenesis was noted

with more advanced disease, and this correlated with

the bone-marrow plasma-cell percentage,

bone-mar-row plasma-cell labeling index, and survival

Role of HHV-8

Human herpesvirus 8 (HHV-8), also known as Kaposi sarcoma-associated herpesvirus (KSHV), was originally described after isolation from a patient with Kaposi sarcoma [51] In these patients

it has been isolated from primary sarcoma cells as well as B cells, macrophages, and dendritic cells HHV-8 has also been shown to be associated with systemic Castleman disease, and primary effu-sion lymphoma where it is localized to just the malignant cells The viral genome encodes a large number of homologs of cellular genes, includ-ing genes functioning in cell regulation (cyclin D), control of apoptosis (bcl-2, death effector domain proteins), cell–cell interaction, immunoregulation, and cytokine signaling, especially IL-6 [52] IL-6, which is considered an important growth factor for myeloma, and a biologically active homolog to human IL-6, termed vIL-6, has been identifi ed in the HHV-8 genome [53] This vIL-6 binds to gp130 directly, suggesting that this molecule may directly activate IL-6R signal transduc tion without binding

to the IL-6R α chain HHV-8 also contains the viral

homolog for interferon regulatory factor (vIRF), which has been detected in patients with multiple myeloma Fibroblasts transfected with vIRF develop into stromal tumors when injected into nude mice, thus suggesting vIRF has properties of a viral onco-

gene [54] Rettig et al have demonstrated vIL-6 RNA

transcripts in cultured KSHV-infected bone-marrow dendritic cells, thus suggesting a role in producing paracrine stimulation of plasma-cell growth and the possibility of transformation of MGUS to MM [55] However, a follow-up study refuted these fi ndings

by using PCR analysis for multiple regions of the HHV-8 genome and serologic studies on patients with MM and found no role of HHV-8 in the etiol-

ogy of MM [56] Ablashi et al performed serologic

assays (whole-virus ELISA) to detect IgG antibody to HHV-8 in 362 patients with MGUS and 110 patients with MM Only 7.8% of the MGUS sera contained HHV-8 antibody to lytic proteins, and no differ-ences were noted in the distribution of antibody

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to HHV-8 in sera from MGUS patients who

pro-gressed to MM The seroprevalence of HHV-8

in MGUS (7.8%), MM (5.4%), and healthy donors

(5.9%) was similar, thus arguing for the lack of

epi-demiologic evidence of HHV-8 in the pathogenesis

of MM Currently, it is unclear if MGUS patients with

HHV-8 infection will progress and go on to develop

overt MM

Relationship between aging and the

development of monoclonal gammopathy

The prevalence fi ndings discussed previously in

the Epidemiology section suggest that there may

be some fundamental changes that occur with the

process of aging that make individuals more

sus-ceptible to developing a monoclonal gammopathy

Animal models have provided some clues in

under-standing the pathophysiology of age-related

mono-clonal gammopathy In aging C57BL/KaLwRij mice,

80% of aged animals will develop a monoclonal

gammopathy that is essentially indistinguishable

from an MGUS in humans [57,58] Also, plasma-cell

dyscrasias such as MGUS, MM, or WM are rarely

seen in C57BL/KaLwRij mice less than two years old

It is hypothesized that these animals may have a

dysregulated immune system that predisposes them

to develop a monoclonal gammopathy Radl has

demonstrated his fi ndings in the C57BL/KaLwRij

mice as an imbalance between a failing T-cell

com-partment (due to an involuted thymus) with an

oth-erwise intact B-cell compartment [57] The loss of

a balanced T-cell/B-cell dichotomy in the immune

system may lead to a restriction of the B-cell

reper-toire and thus to excessive B-cell clonal

prolifera-tion, excessive immunoglobulin producprolifera-tion, and

ultimately to the development of a monoclonal

gammopathy

More recently, Ellis et al have demonstrated that

the relative numbers of the CD30 T-cell subset and

levels of CD30 expression are elevated in activated

lymphocytes from normal aged individuals (60

years) and in MGUS patients, when compared to

younger controls [59] Peripheral blood lymphocytes from MGUS patients and age-matched controls produced comparable levels of IL-6 when activated with anti-CD3 plus IL-2, and costimulation with a soluble form of CD30 ligand (sCD30L/CD8alpha) augmented anti-CD3 inducible IL-6 production similarly in both groups However, peripheral blood lymphocytes from MGUS patients also produced measurable IL-6 when activated with sCD30L/CD8alpha alone This capability was associated with the unique presence of CD30 T cells in the peripheral blood of MGUS patients Furthermore, a higher percentage of activated MGUS T cells express CD30 when activated by incubation with idiotype-expressing autologous serum than those activated

by anti-CD3 plus IL-2 These results indicate that quantitative alterations in CD30 T cells accom-pany aging and MGUS, and that these cells may con-tribute to the chronic activation of B cells though the production of IL-6

In addition to the above murine data, there is also

a wealth of data to indicate that IL-6 gene sion, along with tissue and serum levels of IL-6, all increase with age As indicated before, IL-6 is the chief cytokine implicated in the development of

expres-MM [60–64] Early observations demonstrated an age-associated rise in IL-6 in autoimmune prone mice [63] However, subsequent studies have dem-onstrated a similar age-associated increase in IL-6

in “normal” (without any disease) mice Similarly, an age-associated increase in IL-6 has been described

in healthy older humans and in older adults with coincident age-associated diseases like Alzheimer dementia, osteoporosis, and lymphoproliferative disorders [65] One proposed mechanism for this age-associated increase in IL-6 is the reduced infl u-ence of normally inhibiting sex steroids on endog-enous IL-6 The ability of estrogen to repress IL-6 expression has been studied in human endometrial stromal cells and from observations that menopause

or oophorectomy resulted in increased IL-6 levels [66,67] Similarly, dihydrotestosterone also inhib-its IL-6, albeit to a lesser extent than estrogen [68]

In one study orchiectomy induced bone-marrow

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144 Arati V Rao, Harvey Jay Cohen

IL-6 protein and mRNA expression and led to

increased replication of bone-marrow osteoclast

progenitors, which was prevented by administration

of IL-6-neutralizing antibody or implantation of a

slow-release form of testosterone [69] Thus it seems

likely that at the time of menopause or andropause,

IL-6 gene expression is not that tightly regulated,

leading to inappropriate expression in some

tis-sues and a rise in serum levels This age-associated

rise in IL-6 is of physiologic consequence,

render-ing an individual susceptible to a number of disease

processes induced by pro-infl ammatory signals,

including MM osteoclast stimulation,

lymphopro-liferative disorders, decreased functional status,

and frailty [70] One might hypothesize that in an

older patient with MGUS, the rise in IL-6 levels may

potentiate the usual molecular changes seen in an

aging individual, like decreased immune

surveil-lance, decreased DNA repair, telomere shortening,

and decreased chromosomal stability, and thus lead

A polyclonal increase in the immunoglobulins will produce a broad band that is limited to the gamma region An M-protein may be present even when the total protein concentration, beta- and gamma-globulin levels, and quantitative serum immu-noglobulin levels are all within normal limits A small M-protein may be concealed in the normal

AgeTime

Host resistance

Growth factors

Promotion

MM

Figure 11.2 Relationship between age, HHV-8, and IL-6 in the pathogenesis of MGUS and MM Adapted from Cohen HJ

et al in Balducci L et al., eds, Comprehensive Geriatric Oncology (London: Taylor and Francis, 2004), 194–203 [71].

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146 Arati V Rao, Harvey Jay Cohen

beta or gamma areas and thus easily overlooked

[73] Immunofi xation should be performed when

a peak or band is seen on SPEP, and this confi rms

the presence of and type of M-protein [74,75]

Immunofi xation can detect a serum M-protein of

0.02 g/dL and a urine M-protein of 0.004 g/dL

Urine protein electrophoresis (UPEP) is also

important in the evaluation of monoclonal

gam-mopathy Ideally, immunofi xation of a 24-hour urine

sample is recommended but it can be performed on

a random sample or the fi rst morning specimen It is

not uncommon for a patient to have a normal SPEP

with no M-protein, but for urine immunofi xation

to show a monoclonal light chain [76] Other

stud-ies might include a bone-marrow aspiration and

biopsy, a radiological skeletal bone survey,

quanti-tative serum immunoglobulins, and 24-hour urine

collection for protein quantitation and

electro-phoresis Light chains may not be detected in the

urine because of reabsorption by the proximal renal

tubules Also, there is variation in glomerular fi

ltra-tion and tubular funcltra-tion, and this is relevant in

patients with non-secretory myeloma, solitary

plas-macytoma, or primary systemic amyloidosis These

issues may be circumvented by measuring serum

free light chains, which has been shown to be a very

sensitive method of detecting and monitoring light

chains [77] The quantitative immunoassays can

detect less than 0.1 mg/dL κ and λ chains compared

to 15–50 mg/dL by immunofi xation, and 50–200 mg/

dL by SPEP [78]

It is important to differentiate a patient with

MGUS from one with MM or WM Most patients

with MGUS are asymptomatic, and may be

diag-nosed incidentally by their primary-care physicians

performing a workup for anemia or other related

conditions The size of the M-protein is helpful, and

has been a matter of debate, with the International

Myeloma Working Group using the level of 3 g/dL

as the cutoff value [1] Patients with MGUS do not

have any signs or symptoms from related organ

and tissue damage, as cited above Most patients

with MM or WM have a reduction in polyclonal or

background immunoglobulins, while only 30% of

patients with MGUS have a decrease in polyclonal

immunoglobulins The morphologic appearance of the bone marrow might help in differentiation, and this was illustrated in a study where bone-marrow aspirates from 154 patients were examined by blinded cytologists [79] These patients underwent bone-marrow aspiration as part of a workup for sus-pected myeloma The single morphologic charac-teristic that strongly differentiated MM from MGUS was the presence of large nucleoli in the plasma cells

of patients with MM Higher percentage of plasma cells (mean 48% in MM vs 10% in MGUS), irregu-lar cytoplasmic contour of plasma cells, presence of cartwheel chromatin and vacuolization, more ani-socytosis and plasma cells in clusters were features more prominent with MM bone-marrow specimens

Of note, the popular and commonly used beta-2 microglobulin level is thought not to be useful in differentiating normal individuals from those with MGUS or with early MM [80]

Plasma cells in MM are phenotypically distinct from their normal counterparts, and this has been studied

by fl ow cytometry in patients with MGUS and MM [81] The clonal plasma cells of patients with MGUS show a phenotypic profi le similar to that of myelo-matous plasma cells (CD38, CD56, and CD19), although the proportion of phenotypically normal plasma cells is higher in patients with MGUS than

in those with myeloma Thus, there are actually two populations of plasma cells in persons with MGUS: one is normal and polyclonal (CD38, CD56,CD19), and the other is clonal and has an abnor-mal immunophenotype (CD38, CD56, CD19) This study also demonstrated that the proportion of bone-marrow plasma cells that was polyclonal (as assessed by fl ow cytometry of bone-marrow aspirate with the use of four monoclonal antibodies – CD38

or CD138, CD56, CD19, and CD45) was the best single factor for distinguishing between MGUS and multiple myeloma Only 1.5% of patients with MM had more than 3% of normal plasma cells, whereas 98% of patients with MGUS had more than 3%.Conventional cytogenetics is not very useful in differentiating MGUS from MM, because of the low number of cells in metaphase in MGUS It is now thought that MGUS patients already have

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the chromosomal characteristics of a plasma-cell

malignancy, and this was confi rmed in a study in

which interphase FISH was performed on

bone-marrow plasma cells of 36 patients with MGUS

Chromosomal abnormalities were identifi ed in 53%

patients, with gains in chromosomes 3, 11, 7, and

18 most commonly seen [82] The deletion of

chro-mosome 13q is a clinically relevant feature in MM,

and one study utilized interphase FISH to

demon-strate deletion of the 13q14 locus in 45% patients

with MGUS [83] This was confi rmed by a long-term

follow-up study (median follow-up 30 months)

using conventional cytogenetics and interphase/

metaphase FISH in 18 asymptomatic, untreated

MGUS patients [84] Deletion of 13q14 was

identi-fi ed in identi-fi ve of these patients, and all identi-fi ve progressed

to MM 6 to 12 months after identifi cation of the 13q

anomaly The authors concluded that the extent of

13q deletion does not vary with clinical outcome,

and plays a crucial role in the pathogenesis of MM by

conferring a proliferative advantage to clonal plasma

cells However, these results must be interpreted

with caution since transition from MGUS to MM can

also occur in patients with normal karyotype, as

sug-gested by two patients in the same study

Another tool in distinguishing the two conditions

might be the presence and amount of circulating

plasma cells that can be seen in MM, MGUS, and

smoldering myeloma However, a recent study of 327

patients with MGUS has suggested that patients who

had detectable plasma cells in the peripheral blood

had a shorter median progression-free survival,

shorter median survival, and shorter time to

initia-tion of any therapy for progressive disease [85]

Finally, the plasma-cell labeling index, which

measures the synthesis of DNA, is a useful test for

differentiating MGUS from MM This was

evalu-ated in one study of 80 patients (59 MM, 20 MGUS,

1 plasma-cell leukemia) where plasma-cell

pro-liferation analysis was performed after

bromode-oxyuridine (BRD-URD) incorporation and double

immunoenzymatic labeling on cytological smears

[86] The BRD-URD is incorporated into the nucleus

of cells synthesizing DNA The plasma-cell labeling

index (percentage of cells in S phase) was 0.25 for

MGUS, 0.4 for stage I MM, 2.4 for stage III MM, and 3.7 for plasma-cell leukemia While there was no correlation between the labeling index and beta-2 microglobulin levels between the MGUS and MM patients, there was a correlation of these variables within the different stages of MM

Thus, making the right diagnosis of MGUS is important, as these patients need regular follow-up

in order to detect the development of the nant form, i.e., MM or other related disorders It has been suggested by Kyle & Rajkumar that if a patient has a serum M-protein value of 1.5 g/dL and no other features suggestive of a plasma-cell dyscrasia,(i.e., no anemia, hypercalcemia, renal failure) a bone-marrow examination or skeletal bone survey is not required and an SPEP should be repeated annu-ally [87] Patients with M-protein levels between 1.5 and 2.5 g/dL who are asymptomatic should have additional studies performed, including quantita-tive immunoglobulins and 24-hour UPEP, but do not need a bone-marrow biopsy or skeletal survey The SPEP should be repeated every 3–6 months for

malig-a yemalig-ar malig-and if stmalig-able the durmalig-ation between the tests can be increased to 6–12 months and then annu-ally or if any symptoms occur If the M-protein level

is 2.5 g/dL, complete workup, including titative immunoglobulins, 24-hour UPEP, bone-marrow aspiration and biopsy, and skeletal bone survey, must be performed Given the data we have from recent studies, it might be useful to check serum free light chains in patients with M-protein

quan-1.5 mg/dL and use it along with the kappa/lambda free light chain ratio to risk-stratify patients with MGUS in order to predict progression to more malignant disease (Table 11.1) [88] If the M-protein

is IgM, a bone-marrow aspiration and biopsy is cated to rule out WM or any other lymphoprolifera-tive disorder If all the studies are normal, the SPEP can be repeated every 2–3 months for a year, and if stable can be repeated at 6- to 12-month intervals

indi-It is unusual for a serum monoclonal protein to appear during long-term follow-up Formerly, if the M-protein remained stable for 3–5 years, the process was assumed to be benign and additional follow-up was not mandatory However, the most recent data

Trang 11

dis-148 Arati V Rao, Harvey Jay Cohen

from Kyle et al have demonstrated an average risk

of development of a malignant process of 1% per

year [90] This study found a relative risk of 7.3 for

MM, WM, primary amyloidosis, chronic lymphocytic

leukmia, IgM lymphoma, and plasmacytoma in the

patients with MGUS, as compared to white subjects

in the Iowa SEER registry from 1973 to 1997

Natural history and predictors of

progression

There have been multiple studies conducted on the

natural history of this relatively indolent disease

This chapter, however, will focus on the most recent

studies, with large patient populations and long

durations of follow-up In one study, Baldini et al

fol-lowed 335 patients with MGUS for a median period

of 70 months [91] The frequency of malignant

trans-formation was 6.8%, and there was no difference in

patients with IgM, IgG, or IgA monoclonal protein In

a univariate analysis of the IgG cases, the relative risks

for developing multiple myeloma were as follows: 2.4

for each 1 g/dL increase in IgG serum monoclonal

component, 3.2 for detectable light chain

protein-uria, 4.4 for an increase of one unit log bone-marrow

plasma-cell percentage, 6.1 for age 70 years, 3.6 and

13.1 for a reduction in one or two polyclonal

immu-noglobulins Patients with an M-spike of 1.5 g/dL,

bone-marrow plasma cells 5%, and no urinary light

chains were at lowest risk, and it was suggested in

this paper that patients do not need a skeletal bone

survey or bone-marrow examination until there is

evidence of progressive disease In another study

by Rajkumar et al [89], 1384 patients with MGUS were

followed for 34 years (median 15.4 years) and ing this follow-up MM, lymphoma with an IgM serum monoclonal protein, primary amyloidosis, macroglobulinemia, chronic lymphocytic leukemia,

dur-or plasmacytoma developed in 115 (8%) patients The cumulative probability of progression to one of these disorders was about 10% at 10 years, 21% at 20 years, and 26% at 25 years The overall risk of pro-gression was about 1% per year, and patients were

at risk of progression even after 25 years or more

of stable MGUS Of note, patients with MGUS had shorter median survival than expected for age- and

sex-matched controls (8.1 years vs 11.8 years, p0.001) After analysis of all baseline factors only the concentration and type of monoclonal protein were independent predictors of progression In this study patients with IgM or IgA monoclonal protein had an increased risk of progression as compared to patients

with IgG monoclonal protein (p 0.001), and the risk

of progression was directly related to the tion of monoclonal protein in the serum at the time

concentra-of diagnosis concentra-of MGUS (p 0.001) Finally, a study

by Cesana et al followed 1104 patients with MGUS

for 33 years (median follow-up 65 months) [92] Cumulative transformation probability at 10 and 15 years was 14% and 30% respectively, with 64 patients (5.8%) evolving to multiple myeloma or other related disorders Marrow plasmacytosis greater than 5%, detectable Bence-Jones proteinuria, polyclonal serum immunoglobulin reaction, and a high eryth-rocyte sedimentation rate were independent factors infl uencing MGUS transformation

One very elegant study has hypothesized that the presence of free light chains in the serum, with

Table 11.1 Risk stratifi cation using the size of the serum M-protein and free light chain (FLC) ratio [89].

Absolute risk of progressionRisk group No of patients Hazard ratio at 20 years (%)

Low (serum M-protein 1.5 g/dL and 606 1 7

normal FLC ratio [0.26–1.65])

Intermediate (either risk factor present) 373 3.5 26

High (serum M-protein 1.5 g/dL and 169 6.8 46

abnormal FLC ratio [0.26 or 1.65])

Trang 12

abnormal kappa/lambda ratio, would indicate clonal

evolution in the neoplastic plasma cell and thus

increased risk of progression in MGUS [93] These

investigators utilized a novel, highly sensitive serum

free light chain assay that has recently been

introduced into clinical practice It enables the quantifi

-cation of free kappa and lambda chains secreted by

plasma cells, i.e., those not bound to intact

immu-noglobulin Monoclonal elevations can be reliably

distinguished from polyclonal elevations using the

kappa/lambda ratio The free light chain ratios were

determined in 47 patients with MGUS who had

doc-umented progression to MM or related malignancy,

and compared to 50 patients with MGUS who had

no evidence of progression The presence of higher

kappa/lambda free light chain ratio was associated

with a higher risk of MGUS progression (relative

risk 2.5, 95% CI 1.6–4.0, p 0.001) The same group

has now utilized the kappa/lambda free light chain

ratio along with the serum M-protein level in 1384

patients with MGUS to risk-stratify these patients

and thus predict the absolute risk of progression at

20 years (Table 11.1) An abnormal kappa/lambda

free light chain ratio of 0.26 or 1.65 was

consid-ered a major independent risk factor for progression

of MGUS to MM or other related malignancy

Thus, in general MGUS can be characterized as

a pre-malignant condition with a limited and

con-trolled lymphoplasmacytic expansion and a benign

course However, transformation to a more

malig-nant condition like MM can occur, and generally is

associated with higher tumor burden and a

progres-sively downward course

Summary

Monoclonal gammopathy of unknown signifi cance,

a pre-malignant condition, is the most common

plasma-cell dyscrasia, and its incidence increases

with age It is characterized by a serum M-protein of

3.0 g/dL or trace or no light chains in a 24-hour

urine collection, less than 10% plasma cells in the

bone marrow, and the absence of lytic bone lesions,

anemia, hypercalcemia, and renal insuffi ciency

There is a clear association between aging and the development of monoclonal gammopathy, and this may be due to increased levels of IL-6 and other related cytokines Long-term follow-up studies have demonstrated the rate of transformation of MGUS

to MM or related disorder to be 1% per year Risk factors for progression have been studied by multi-ple groups, and those that place the patient at high risk include size of M-protein, presence of urinary light chains, and bone-marrow plasmacytosis More sophisticated tests like the plasma-cell labeling index, and FISH to detect 13q deletion, might also be helpful in determining risk of transformation More recently, very sensitive assays for serum free light chains have been developed, enabling us to calculate the kappa/lambda chain ratio and use this to predict progression of MGUS to more serious MM While there are no absolute fi ndings at the time of diagno-sis of MGUS that allow us to distinguish patients who will remain stable from those who will develop more malignant disease, we do have many diagnostic tests

in our armamentarium to help follow these patients

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Anemia of aging

Trang 20

Introduxtion

Oswald StewardReeve-Irvine research center, departments of anatomy & nurobiology, nurobiology & behavior,

and neurosurgery, university of california at irvine, Irvine, CA 92697

Introduction

Anemia represents a common problem among

the elderly, with a prevalence of 5–10% for the

community-dwelling elderly between 65 and 74

years, and over 20% for seniors 85 years and over In

hospitalized elderly patients and in skilled nursing

facilities, anemia prevalence ranges from 40 to 50%

The recent appreciation of the numerous adverse

consequences of anemia has generated interest in

a more complete understanding of anemia in the

elderly Erythropoietin (EPO) is a hormone central

to the regulation of red-blood-cell production that

increases in response to falling hemoglobin

con-centration Paradoxically, although EPO levels rise

slightly with age in non-anemic elderly people,

the expected EPO response to anemia appears

sig-nifi cantly blunted in the elderly, supporting a

rela-tive endogenous EPO defi ciency This relarela-tive EPO

defi ciency, possibly attributable to occult renal

insuffi ciency, may play a central role in the rising

prevalence of anemia with advancing age and

unex-plained anemia in the elderly

Erythropoiesis

Hematopoiesis, the production of blood elements,

occurs in an orderly hierarchical fashion

Mainte-nance of mature peripheral blood cells (i.e.,

plate-lets, red blood cells or erythrocytes, neutrophils,

eosinophils, basophils, monocytes, lymphocytes,

natural killer cells, and dendritic cells) demands

Erythropoietin and aging

Andrew S Artz

ongoing production to meet losses and respond to stresses A pluripotent hematopoietic stem cell pro-duces committed progenitors of myeloid, erythroid, and megakaryocytic lineages Erythropoiesis denotes the process of forming mature red blood cells The earliest erythroid lineage progenitors include the BFU-E (burst-forming unit, erythroid), which later give rise to CFU-E (colony-forming unit, erythroid) Normal erythropoiesis in adults occurs exclusively in the bone marrow

Erythropoietin physiology

Various hematopoietic growth factors support proliferation, differentiation, and survival of stem cells Erythropoietin, a glycoprotein hematopoietic growth factor composed of 165 amino acids after modifi cation from the 193-amino-acid molecule encoded by mRNA, serves as a primary regulator of red-cell production [1,2] The markedly elevated cir-culating EPO levels in patients with aplastic anemia allowed initial EPO isolation from the urine [3] The glycosylation present on EPO delays clearance of the molecule, but is not necessary for biologic activ-ity [4] Synthesis and EPO regulation occur prima-rily through the kidney, from interstitial fi broblasts and/or from proximal tubular cells, with a smaller contribution by liver hepatocytes [5–9] As a con-sequence, renal failure inexorably leads to anemia from impaired EPO production

Under normal homeostasis, suppressed tion of EPO leads to a minimal constitutive pro-duction Reduced tissue oxygenation (rather than

transcrip-Blood Disorders in the Elderly, ed Lodovico Balducci, William Ershler, Giovanni de Gaetano

Trang 21

158 Andrew S Artz

diminished red-cell production), typically from

anemia or hypoxia, potently stimulates synthesis

with logarithmically elevated serum EPO levels [10]

Hypoxic conditions promote activation of the hypoxia

inducible factor 1 (HIF-1) pathway, with expression

of the alpha and beta subunits, and eventual

down-stream EPO gene transcription [11] With reduced

oxygen delivery, the number of renal cells producing

EPO may also increase [9] Elevated serum EPO levels

enhance erythrocyte production primarily by

inhib-iting apoptosis of erythroid progenitor cells, although

EPO also promotes erythroid progenitor proliferation

and differentiation [12]

Erythropoiesis with aging

Early laboratory models helped guide subsequent

human studies describing EPO kinetics with aging

Basal erythropoiesis and red-cell mass in aged mice

appears similar to younger mice, although measured

hematocrit may be slightly lower secondary to

dilu-tion [13,14] In response to stressors, such as hypoxia,

bleeding, or environmental perturbation, aged mice

have an impaired erythropoietic response [13,15,16]

Human studies in older subjects with unexplained

anemia have shown normal BFU-E cells but

dimin-ished CFU-E, the erythroid precursors most replete

with EPO receptors, suggesting a block in

differenti-ation possibly mediated by inadequate endogenous

EPO levels Further, CFU-E viability requires EPO [17]

Erythropoietin receptors

Hematopoietic growth factors in general bind with

high affi nity to specifi c receptors on target cells

EPO activity necessitates binding to erythropoietin

receptors (EPO-R) Within the marrow

compart-ment, EPO receptors (EPO-R) occur on the two

classes of committed erythroid precursors, the

BFU-E and CFU-BFU-E [18] While BFU-BFU-E cells have a high

proliferative capacity, they only have low EPO-R

expression The CFU-E cells, progeny of the BFU-E

cells, have a lower proliferative capacity but increased

EPO-R density After EPO attaches to EPO-R, signal

transduction permits cell survival The heterogeneity

of EPO sensitivity at the molecular level of such cells may also facilitate erythropoietic regulation [19]

Non-marrow activity of erythropoietin and erythropoietin receptors

Recognition of EPO in non-hematopoietic tissue has pointed to the potentially pleiotropic effects of the hormone The presence of EPO-R on vascular endothelial cells [20], various renal cells [21,22], human tumor cells [23], and the central nervous system (CNS) refl ect a multiplicity of non-hematopoietic roles for EPO, and possibly therapeutic implications Research into the understanding of EPO in CNS development and homeostasis has been particularly fruitful Similar to the bone-marrow compartment, hypoxia-regulated EPO and EPO-R expression occur within the central nervous system [24–27] Endogenous EPO does not signifi cantly cross the blood–brain barrier [28], although large pharmacologic doses may pene-trate the CNS EPO may protect the CNS after hypoxic injury [29], motivating studies of pharmacologic EPO

as neuroprotective therapy after strokes, with ing preliminary results [30] Changes with advancing age have not been studied

promis-Normal erythropoietin response

to anemia

Serum endogenous EPO levels typically remain within a narrow reference range for young healthy patients at normal hemoglobin concentrations Isolation of extremely high concentrations of EPO from the urine of patients with aplastic anemia helped uncover the dynamic nature of EPO regula-tion in response to anemia The advent of immu-noassays to measure serum EPO further promoted research clarifying serum EPO kinetics [31]

Relative erythropoietin defi ciency

Determining the expected rise in serum EPO to anemia remains problematic and complicates both

Trang 22

clinical and research data interpretation Reference

ranges reported by laboratories describe serum EPO

levels from non-anemic healthy populations and do

not refl ect the dynamic nature of EPO responses

Interpretation of these values may result in

con-fusion, as a serum EPO level within the reference

range (approximately 4–28 mIU/mL, depending on

the laboratory) for an anemic patient often

signi-fi es a relative EPO designi-fi ciency Multiple methods of

quantifying EPO responsiveness have further

com-plicated evaluating studies, including mean and

median EPO levels, with and without adjustment for

hemoglobin concentration Generating a standard

curve documenting the expected serum EPO for a

given hemoglobin concentration enables

appropri-ate statistical modeling but also requires logarithmic

transformation of EPO levels Such transformations

further complicate clinical interpretation

Among younger subjects with signifi cant

ane-mia, a normal EPO response has been characterized

as a semilogarithmic increase In general, serum

EPO levels inversely correlate with hemoglobin, at

least with signifi cant degrees of anemia in younger

subjects experiencing blood loss, iron defi ciency,

hemolysis, or erythroid marrow aplasia Assuming

this represents the normal kinetics of serum EPO in

response to anemia, a curve of the appropriate EPO

concentration for the degree of anemia has been

proposed [32,33] Lack of precision represents a

major obstacle to comparing EPO levels to this

“nor-mal curve.” For example, it remains unknown what

EPO concentration below the predicted level should

be considered low Nevertheless, the predicted EPO

level for a given hemoglobin concentration may

approximate an appropriate EPO response and

per-mit defi ning a relative EPO defi ciency

Renal failure exemplifi es the prototypical

ane-mia from impaired EPO production In many other

conditions, EPO levels are elevated above the

ref-erence range for non-anemic controls but remain

signifi cantly below the expected concentration For

example, an EPO level of 50 mIU/mL at a

hemo-globin of 9.0 g/dL, although far above the reference

range, would still be considered a suboptimal EPO

response This inadequate EPO response for the

degree of anemia commonly occurs with tions such as cancer, HIV, and rheumatoid arthritis [34–36] The mechanism may be multifactorial, but it

condi-is likely that there condi-is a contribution from reduced renal EPO secretion and impaired marrow responsiveness

Erythropoietin levels in the elderly

Non-anemic elderly

Multiple studies have assessed endogenous serum EPO levels in older subjects without anemia (Table 12.1)

Japanese older subjects living in a “home for aged” were compared to younger healthy controls [37] The non-anemic older subjects had numerically but statistically non-signifi cant higher mean EPO levels (mean EPO of 20.4 mIU/mL and 15.7 mIU/

mL for older and younger, respectively) In a study

of older adults with a mean age of 77.8 years ing in a home for the elderly, Kario and colleagues ascertained serum EPO levels [39] An elevated mean EPO level of 26.9 mIU/mL ( 15.2) was observed among the 116 non-anemic elderly, compared to

resid-a meresid-an of 15.8 mIU/mL in younger controls Kresid-ario

et al subsequently reported higher EPO levels

among non-anemic elderly (mean EPO of 24.3 mIU/mL) compared to younger subjects (mean EPO of 14.8 mIU/mL) [40] Also, they showed a signifi cant inverse relationship between EPO and hemoglobin (Hb) concentrations in non-anemic elderly subjects

(r  –0.302, p  0.001) Further, a signifi cant

associa-tion existed between advancing age and rising EPO

(r  0.220, p  0.01) These studies provided early

evidence for mildly increased EPO occurring with advancing age and/or slight reductions of hemo-globin concentration, even within the normal range.Powers and colleagues found EPO levels in

25 healthy elderly people, with an age range of 60–82 years, to be similar to those in younger con-trols (means of 10.8 and 13.1 mIU/mL, respectively) [38] In a larger cohort, the same author showed similar EPO levels in younger and older subjects, after adjusting for hematocrit [41] Joosten and

Trang 23

160 Andrew S Artz

colleagues quantifi ed endogenous EPO levels in

hospitalized non-anemic elderly patients with a

mean age of 81.5 years and mean hemoglobin of

14.2 g/dL, and compared them to young controls

with a mean hemoglobin of 14.6 g/dL [42] They

showed mean EPO levels did not differ by age

cohort, with levels of 7.5 and 9.5 mIU/mL for older

and younger subjects, respectively These studies by

Powers and Joosten appear contradictory to earlier

data on rising EPO levels with age in those without

anemia However, Powers’ subjects primarily

car-ried a diagnosis of rheumatoid arthritis, a condition

known to blunt EPO responses, and the participants

in the study by Joosten were hospitalized patients,

presumably having other comorbid conditions also

blunting EPO responsiveness Thus, one could not isolate age-related changes in EPO responsiveness

To identify the diurnal variations in endogenous EPO, Pasqualetti and colleagues measured serum EPO in 20 younger adults with a mean age of 45 years and mean hemoglobin of 13.9 g/dL and compared them to older adults with a mean age of 65 years and mean hemoglobin of 13.5 g/dL [46] In general, EPO levels peaked around 6 pm and were at their lowest around midnight Young and old subjects exhibited

a similar diurnal variation, although younger

sub-jects had higher mean daily levels (p 0.05) and

more diurnal variation (p 0.05) in EPO relative

to older subjects The signifi cance of these fi ndings remains unclear, although one may postulate that

Table 12.1 EPO levels in non-anemic elderly adults compared to younger adults.

Mori et al 1988 [37] n  78 (13 with chronic disease, n  127 Elderly mean EPO 20.4 10.4

16 with cancer, 10 without Age 22–46 years Younger 15.7 1.3etiology), home for the aged

Age 70–89 years

Powers et al 1989 [38] n 25 n 30 Elderly mean EPO 10.8 6.4

Age 60–82 years Age  65 years Younger 13.1 5.5

Kario et al 1991 [39] n 116 n 26, volunteers Elderly mean EPO 26.9 15.2

Mean age 77.8 years for all Mean age 42.4 Younger 15.8 EPO rose with age elderly

Kario et al 1992 [40] n 150, ambulatory n 111 Elderly mean EPO 24.3 (17.2–34.5) Age  60 years Mean age 46 years Younger 14.8

Age-related increase in EPO

Nafziger et al 1993 [41] n 30 Not provided Elderly mean EPO 11 3.5

Levels “similar” to non-anemic

Joosten et al 1993 [42] n 27, hospitalized n 30, hospital staff Elderly mean EPO 7.5

Mean age 81.5 years Mean age 32.5 years Younger mean EPO 9.5

Goodnough et al n 31, autologous blood n 40, from same Elderly with higher EPO using

1995 [43] donors prior to surgery population Log EPO and age

Mean age 71.1 years Age  65 years, mean No difference in EPO in subset of

47.6 years 18 patients

Ershler et al 2005 [44] n 143, healthy elderly None Mean EPO 13.0 and rose with

EPO, serum endogenous erythropoietin level (mIU/mL).

Trang 24

more tightly regulated EPO in the young resulted in

stronger temporal changes

Endogenous EPO response to anemia

in the elderly

As previously stated, determining a normal EPO

response to hemoglobin changes represents a major

challenge in accurate characterization of EPO

kinet-ics Clearly, young subjects with hemolytic anemia or

subjected to phlebotomy have a brisk rise in serum

EPO levels An absolute EPO defi ciency causes

ane-mia in renal failure, and a relative EPO defi ciency

contributes to anemia in various conditions such as

cancer, autoimmune conditions, prematurity, and

human immunodefi ciency Studies evaluating serum

endogenous EPO in anemic elderly subjects have

allowed a better understanding of changes in serum

EPO and erythropoiesis with aging

Studies in anemic elderly

Mori et al examined iron-defi cient anemic older

subjects from ages 70 to 89 years, and found elevated

endogenous EPO levels with lower hemoglobin

concentrations [37] The preserved EPO

respon-siveness in elderly iron-defi cient anemic subjects

indicated that age does not necessarily confer an

absolute EPO defi ciency

In one of the earliest studies describing a relative

EPO defi ciency in the elderly, Carpenter and

col-leagues detailed hematologic parameters and EPO

concentration for a large group of younger and older

subjects (aged 70 years and over) [47] EPO levels

rose among the iron-defi cient elderly with

worsen-ing anemia In contrast, for the elderly with

normo-cytic anemia (n 375), the EPO response to anemia

was reduced relative to the elderly with microcytic or

macrocytic anemia and relative to all iron-defi cient

anemic subjects Similarly, Kario and others reported

an inverse semilogarithmic relationship between

EPO and Hb concentrations in subjects with Hb

con-centrations less than 12.0 g/dL (r  –0.559, p  0.001)

EPO concentrations in the elderly were lower than

those in young subjects with iron-defi ciency anemia

with the same Hb level [39] In a follow-up study, the

same group showed elevated EPO levels with falling hemoglobin, but this was restricted to the elderly with iron-defi ciency anemia The authors suggested that while elderly subjects with iron defi ciency may have a degree of preserved EPO responsiveness

to falling hemoglobin, anemia of advancing age is characterized by a relative EPO defi ciency, and they hypothesized that pharmacologic EPO replacement may reverse the defi cit [40]

Joosten and others investigated hospitalized older subjects and found that among 24 iron-defi cient anemic older persons with a mean age of 83 years and mean hemoglobin of 10.0 g/dL, lower hemo-globin correlated with higher EPO levels [42] How-ever, in the older subjects with anemia due to chronic disease or acute infection, the lack of correlation between EPO and hemoglobin concentration sug-gested a blunted EPO response Whether conditions characterized by infl ammation or aging in this hos-pitalized cohort blunted the EPO response could not

be determined

Among 31 iron-defi cient elderly subjects, Nafziger and colleagues observed the expected inverse correlation, with lower hemoglobin and higher EPO concentrations [41] However, com-pared to 33 subjects less than 60 years of age, the correlation of rising EPO with lower hemoglobin values was less pronounced Moreover, there was

a trend towards lower EPO levels in older defi cient anemic patients relative to younger anemic subjects, reaching signifi cance when hemo-globin concentration fell below 10 g/dL, suggesting that iron defi ciency and severe anemia represent additional stimuli to EPO secretion that remain sub-optimal in the elderly

iron-In a study of older Japanese subjects compared

to patients under 60 years of age, Matsuo and leagues enumerated EPO levels and the immature reticulocyte count, a measure of marrow erythro-poietic response, among iron-defi cient subjects [48] Higher reticulocyte counts refl ect more brisk erythropoiesis Although median EPO levels did not differ in the young and old, the reticulocyte count/log EPO concentration ratio was 3.3 for older

col-and 8.1 for younger subjects (p 0.01), suggesting

Trang 25

162 Andrew S Artz

a diminished marrow response to circulating

endogenous EPO levels among the elderly The lack

of adequate iron for red-cell synthesis confounds a

reliable determination of the marrow

erythropoi-etic responsiveness to endogenous EPO secretion

Alternatively, fewer iron stores or reduced capacity

with aging to mobilize iron may have accounted for

reduced erythropoiesis

Pieroni and colleagues studied anemia of chronic

infl ammation in 56 older subjects with cancer,

infec-tion, or another infl ammatory disorder, and

com-pared them to a control group of older subjects In

general, they observed blunted EPO

responsive-ness to anemia, until moderate to severe anemia of

8.5 g/dL [49] The underlying conditions may also

have impaired EPO responsiveness, resulting in

blunted EPO levels Nevertheless, these data, in

con-cert with the study by Joosten and colleagues [42],

demonstrate that markedly low hemoglobin levels

provide a stronger EPO stimulus

Kamenetz and colleagues evaluated 17 elderly

sub-jects with unexplained anemia, ranging in age from

65 to 90 years, and compared them to older

non-anemic controls and subjects with anemia and recent

stroke [50] Anemia was defi ned as a hemoglobin

con-centration 12 g/dL EPO levels did not signifi cantly

differ across the three different groups, refl ecting

impaired EPO responsiveness in the anemic patients,

as one would expect elevated EPO levels in anemic

subjects

In a study of 71 healthy blood donors undergoing

aggressive phlebotomy, Goodnough and colleagues

characterized the endogenous EPO response [43]

Older subjects had similar EPO responsiveness

to phlebotomy Red-cell expansion in response to

phlebotomy showed a non-signifi cant trend for

being reduced in the elderly (p 0.10)

In a case series of patients referred to a

hematol-ogy practice, seven patients were described with

unexplained anemia who later received

pharma-cologic therapy for the anemia At baseline,

hemo-globin and endogenous EPO were 10.0 g/dL and

10.8 mIU/mL, respectively, providing evidence for

a blunted EPO response in patients primarily with

unexplained anemia [51]

We analyzed 900 elderly subjects drawn from fi ve nursing homes to determine anemia prevalence [52] Eighty-one subjects with anemia or borderline low hemoglobin from the cohort were randomly selected for further characterization [53] Among 60 patients with confi rmed anemia, iron defi ciency and unex-plained anemia accounted for 23% and 45% of cases, respectively (Table 12.2) Correlating serum EPO lev-els to hemoglobin concentration revealed increased EPO levels with iron defi ciency but no correlation for seniors with unexplained anemia (Fig 12.1) The rise

in EPO among those with iron defi ciency appeared less than expected, suggesting that a blunted EPO response, albeit to a lesser degree than in unexplained anemia, existed with iron defi ciency In the anemic residents with adequate renal function (glomerular

fi ltration rate 30 mL/minute and serum creatinine

2.0 g/dL), calculated creatinine clearance remained independently associated with lower EPO levels when

controlling for age, gender, and Hb value (p 0.008), suggesting that even mild renal insuffi ciency promotes impaired EPO responsiveness

Although a relative EPO defi ciency for the degree of anemia has been described for numerous conditions, such as cancer, prematurity, human immunodefi -ciency, autoimmune conditions, and the intensive care unit, limited data assess the impact of older age

in these diseases on endogenous EPO ness In a small cohort of 20 cancer patients aged

responsive-70 years and over undergoing platinum-based therapy, Cascinu and others reported mean endog-enous EPO levels similar to those in patients younger than 70 years (51 and 62 mIU/mL, respectively) [54] Absent correlations of serum EPO for hemoglobin, limited inferences regarding age can be made

chemo-EPO responsiveness in the elderly with anemia: conclusions

The studies of anemic older patients permit several conclusions A relative EPO defi ciency characterizes the anemia associated with aging (unexplained ane-mia) Chronic disease does not adequately explain the blunted responses, suggesting EPO defi ciency may be a feature of advancing age and/or occult

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