Posters / Leukemia Research 35 (2011) S27–S142 S81

CD19+ cell surface marker. In blood, the decreased count of B cells

was found in RA+RCMD, RARS and 5q− patients (p < 0.01). In

MDS high risk marked variation of B cell count was observed.

The longtime follow up of several patients with low number of

B cells showed good reproducibility. Due to the determination of B

lymphocytes clonality (according to % of kappa and lambda in CD19)

we were able to exclude those MDS patients with simultaneous

latent B-CLL or benign monoclonal lymphocytosis. Surprisingly, the

decrease of B cells in low risk MDS patients was not connected with

unfavorable prognosis. In bone marrow B cell values oscilated and

no statistical difference in comparison with controls was found.

While studying gene expression profiles in CD34+ cells of 51 MDS

and 7 controls we identified a group of l3 genes with decreased

expression (POU2AF1, VPREB1, VPREB3, CD79A, EBF1, LEF1, BCL3,

IRF8, IRF4, BCL6, KLF6, TCF3 and RAG1). These are genes involved

in B cell development or activation. Furthermore, microRNAs were

studied in CD34+ cells of 39 MDS patients and 6 controls by

microarray analysis. Overexpression of miR-127 from the cluster

14q32 were ascertained. These microRNAs are involved in B-cell

differentiation. The B cell decrease is not a matter of age. However,

premature apoptosis of B cells in MDS cannot be excluded.

Conclusion: B cell decrease is encountered especially in low risk

MDS and is a constant finding in these patients. Decreased gene

expression and overexpression of microRNAs involved in B cell

development may contribute to low B lymphocyte count in MDS.

Supported by MSM 0021620808

207

Characterization of FMNL1 expression in peripheral blood and

bone marrow cells of MDS patients

M.R. Lopes1, P. Favaro1,2, F. Traina1, P. de Melo Campos1, J. Machado-

Neto1, I. Lorand-Metze1, F.F. Costa1, S. Olalla Saad1. 1Hematology and

Hemotherapy Center – University of Campinas/Hemocentro – Unicamp,

Instituto Nacional de Ciencia e Tecnologia do Sangue, Campinas,2Department of Biological Sciences, Federal University of Sao Paulo,

Diadema, Brazil

Myelodysplastic syndromes (MDS) are a heterogeneous group of

malignant stem cell disorders characterized by dysplastic and

ineffective blood cell production. Evidences have shown that T-cell

mediated marrow suppression is the cause of the cytopenia

in approximately 20–30% of MDS patients. FMNL1 belongs to

a conserved family of formin-related proteins, indispensable for

many fundamental actin-dependent processes. Human FMNL1 is

restrictedly expressed in hematopoietic cells, including lymphocytes,

and overexpressed in chronic lymphocytic leukemia and lymphoma

samples as well as in malignant cell lines. Moreover, FMNL1

participates in the regulation of the cytoskeleton in activated T

lymphocytes and the control of the polarization of centromeres,

required for cytotoxic function of these cells. The aim of this work

was to characterize FMNL1 expression in peripheral blood CD3+ and

in total bone marrow cells of patients with MDS and normal donors

and to compare FMNL1 expression level between low-risk, high-risk

MDS and normal donor cells. A total of seventy-one patients with

a diagnosis of MDS, receiving no treatment, from the Hematology

and Hemotherapy Center (Hemocentro), at UNICAMP, were included

in the study; twenty-two samples from normal donors were used

as controls. Patients were grouped into low-risk and high-risk

disease, as RA/RARS (40/8) vs. RAEB/RAEBt (17/6) (FAB classification),

and 0+1 cytopenia (n = 31) vs. 2+3 cytopenia (n = 40) (according to

number of cytopenias). This study was approved by the National

Ethical Committee Board. FMNL1 expression levels from CD3+ cells

(obtained by Ficoll-Hypaque followed by magnetic selection) or total

bone marrow cells were determined by quantitative PCR (q-PCR).

FMNL1 expression was significantly higher in MDS CD3+ peripheral

lymphocytes when compared with normal donor cells (P=0.01).

There was no statistical difference between low-risk and high-

risk MDS group CD3+ cells. In the bone marrow samples, FMNL1

expression was higher in low-risk compared to high-risk MDS

according to FAB (P =0.07) and number of cytopenias (P =0.01).

During the early stages of MDS, one mechanism contributing

to hypercellular marrow and peripheral blood cytopenia is the

significant increase in apoptosis in haematopoietic cells. The higher

expression of FMNL1 in MDS peripheral lymphocytes and in low-

risk bone marrow may be related to a clonal or oligo-clonal T cell

expansion. Further studies will be necessary to test this hypothesis.

Supported by FAPESP, CNPq and Instituto Nacional de Ciencia e

Tecnologia do Sangue-INCT do Sangue.

208

Iron chelation therapy with deferasirox in transfusion

dependent myelodysplastic syndrome patients. Preliminary

report from the prospective MDS0306 GIMEMA trial

E. Angelucci1, A.A. Di Tucci1, S. Storti2, D. Magro3, P. Tosi4,

S. Amadori5, P. Leoni6, M. Gobbi7, M. Brugiatelli8, F. Pane9,

G. Visani10, F. Nobile11, F. Lauria12, R. Fanin13, G. Specchia14,

P. Ditonno14, G. Rossi15, G.L. Forni7, A. Abbadessa16, A. Olivieri17,

S. Porcedda1, F. Pilo1, A. Piciocchi A18, M. Vignetti18, S. Tura19.1Haematology, Businco Hospital, Cagliari, 2Haematology, Campobasso,3Haematology, Catanzaro, 4Haematology, Rimini, 5Haematology, Roma,6Haematology, Ancona, 7Haematology, Genova, 8Haematology, Messina,9Haematology, Napoli, 10Haematology, Pesaro, 11Haematology, Reggio

Calabria, 12Haematology, Siena, 13Haematology, Udine, 14Haematology,

Bari, 15Haematology, Brescia, 16Haematology, Caserta, 17Haematology,

Potenza, 18GIMEMA Data Centre, Rome, 19Haematology, Bologna, Italy

The recent development of a safe and efficient once daily oral

iron chelator (Deferasirox, Exjade) made possible regular chelation

therapy in transfusion dependent MDS patients. However the

reported clinical experience is limited to selected populations. For

this reason the GIMEMA group developed a phase IIIb prospective

trial to test safety and efficacy of Deferasirox in a large population

of patients comparable to general MDS population.

One hundred and fifty-nine transfusion dependent IPSS low-

intermediate1 risk MDS patients were enrolled. Baseline

characteristics were (data are expressed as median unless otherwise

specified): age 72 years (range 24–87); 48 patients IPSS low risk

and 75 Intermediate1; duration of transfusion dependency before

treatment 20 months (12–36) corresponding to 38 (22–70) packed

red blood cells transfusions received. Baseline serum ferritin was

2000ng/ml (1471–3000). Baseline Charlson and CIRS comorbity

scores were 1 (0–1) and 0.2 (0.1–0.4), respectively. Patients started

treatment with the standard 20mg/kg Deferasirox dose but dose

adjustments on clinical indications were allowed.

Sixtyone patients (49%) prematurely interrupted the study (drop

out), 62 (51%) patients completed the planned year of treatment.

Serum Ferritin evolution in the 62 patients who completed the

protocol showed a statistically significant decrement during the 12

months follow up (P < 0.001, median value from 2000 to 1500ng/ml).

In multivariate model for drop out rate, high Charlson co-morbidity

score >1 (Odds Ratio 1.45) and duration of transfusion dependency

before chelation treatment < 24 months (Odds Ratio 0.97) were

significant risk factors for drop out. Drops out were mainly related

to progression to acute leukemia (10 patients), MDS clinical problem

(20 patients), unrelated causes mainly aging related (12 patients)

and 6 by others causes. Drug related toxicity was drop out cause in

13 patients (11% of the entire population). Main causes of toxicity

related drop out were increase of creatinine and gastro-intestinal

disturbance.

Preliminary results from the GIMEMA MDS0306 study confirmed

feasibility of Deferasirox therapy in transfusion dependent MDS

patients. High Charlson co-morbidity score and duration of

transfusion dependency before chelation treatment were a

significant risk factors for drop out. Toxicity related drop out and

severe side effects were similar to those reported in other trials

even if the present population presented clinical characteristics

S82 Posters / Leukemia Research 35 (2011) S27–S142

of more advanced disease and age. Serum ferritin behavior

confirms Deferasirox efficacy. Data indicate that thalassemia derived

Deferasirox schedule could be not appropriate to elderly MDS

patients.

clinicaltrial.gov identifier NCT00469560

209

A retrospective analysis using 13-cis retinoic acid (13CRA) and

alpha tocopherol (AT) in MDS patients to prevent progression

E.C. Besa. Medical Oncology, Kimmel Cancer Center, Thomas Jefferson

University, Philadelphia, PA, USA

Multiple studies have been done involving the use of 13-cis retinoic

acid, by itself and in, combination with other drugs in patient

with MDS. Several reports show a disappearance of cytogenetic

abnormalities and clinical remission. However a randomized blinded

control study comparing placebo with high doses of 13-cis retinoic

acid failed to show any advantage of 13-cis retinoic acid over placebo.

But this study had a great deal of patients discontinue therapy

secondary to toxicity that was caused by the of 13-cis retinoic acid,

this may have contributed to the failure of the study to show any

advantage. Subsequent studies have shown that the beneficial effects

of 13-cis retinoic acid may be seen only in the early stages of the

disease, possibly in the low risk category per IPSS. The study by

Clark, et al. [8] randomizing MDS patients into 2 groups. The INT-2

and High risk MDS patients were randomized between high dose 13

CRA vs low dose Ara-C did not show any difference in response or

survival. However, in low risk MDS specially in RA group, the arm

treated with low doses of 13 CRA at 40mg/day demonstrated an

overall improved survival compared to placebo or supportive care

alone. Because of our observation that alpha tocopherol (AT) at doses

of 400–800 units per day alleviated skin toxicity of 13 CRA, we then

proceeded to treat a cohort of MDS patient with the combination. We

compared a group using high doses of 13 CRA at 160–200mg/day for

6 months (HDSD) in 29 patients versus using low doses of 40mg/day

until progression (LDLD) on 20 patients both with 800 units of AT.

Both groups were similar in age (mean, range) 66.2 (28–84) vs

69.6 (25–93) years, male/female ratio of 16/13 vs 11/9, duration

from diagnosis to treatment of 13.5 vs 14.5 mo. IPSS scores and

transfusion requirement. Responses were observed in both groups

overall (CR, PR and SD) 44.8% in HDSD and 75% in LDLD with similar

AML transformation in INT-1–2 of 2%. A better median survival was

observe with 5 patients still alive at 46 months in LDLD group

compared to 27.5 months in HDSD group with a difference of 18mo

(1.5 years). This suggest a lack of toxicity and good tolerance using

13CRA ar 40mg/d with AT for long term preventive measure in early

phase MDS may result in prolonged survival.

210

Should immunosuppressive therapy (IST) be used more often

in lower risk MDS?

S. Cereja1, S. Brechignac1, L. Ades1, T. Braun1, S. Boehrer1, Z. Hebibi1,

R. Sapena2, O. Beyne-Rauzy3, N. Vey4, H. Dombret5, D. Bordessoule6,

F. Dreyfus2, P. Fenaux1, C. Gardin1. 1Hopital Avicenne/Universite

Paris XIII, Bobigny, 2Hopital Cochin, Paris, 3Hopital Purpan, Toulouse,4Institut Paoli Calmettes, Marseille, 5Hopital Saint Louis, Paris,6CHU Limoges, Limoges, France

Aim: In low-int-1 risk MDS patients, immunosuppressive therapy

with ATG+/−CsA may yield a 30 to 50% erythroid durable response

rate, including red cell transfusion independence. Predictors of

response to IST in MDS include low/Int-1IPSS (especially int-1

without excess blasts due to presence, in addition of anemia, of

thrombocytopenia or intermediate (int) karyotype) and age ≤60y

and HLA DR-15 positivity. We reviewed the characteristics and

outcome of MDS patients treated with IST in our centre and assessed

the frequency of low/int-1 MDS patients potentially candidates to

IST, based on published prognostic factors, in our GFM patient

database.

Methods: In 6 years, we treated 12 MDS by IST. From mid 2003 to

end of 2008, 1311 MDS patients were entered into the GFM registry.

Results: The 12 patients treated with IST included 8M/4F. Median age

was 63.5 y, (50%pts older than 65y). WHO classification was RA=3,

RCMD=7, RAEB1=2; cytogenetics normal/−Y =10; +8 =1; failed =

1pts). IPSS was Int-2 in 1, Int-1 in 10 and ND in 1pt. All pts were

RBC transfusion dependent and 11/12 also had platelets ≤50G/L. Pts

received ATG (n =7) or ATG+CsA (n =5). Five (42%) pts, all with RA

or RCMD, were responders at 6 months, (4 RBC-TI including 3 CR

and one HI-E+ HI-P) and 7 were non-responders 4/6 DR-15 positive

pts responded versus 1/5 DR-15 negative pts. Three of the 6pts aged

>65 y responded. With a median follow-up of 21.5 months erythroid

response was ongoing after 17, 21, 44, 47 and 62months, respectively.

Three NR pts died at 11, 16 and 18 months. In the GFM database, pts

with transfusion dependent anemia and IPSS low or int-1, and IPSS

int-1 with <5% marrow blasts represented respectively 20 and 5.5%

of the total pt number considering only pts aged <60y, 22 and 6.5%

considering pts <65y, and 24 and 6.5% considering pts <70y.

Conclusions: We confirm that durable erythroid responses to IST

can be obtained in selected int-1 MDS patients. In our database

the proportion of potential candidates to IST however was very

low whatever the age of patients. Outpatient therapy with low-

dose alemtuzumab may extend the use of IST in older patients, if

confirmed to have a lower toxicity compared to ATG+/−CsA.

211

Deferasirox vs. deferiprone treatment in iron overloaded

patients with myelodysplastic syndrome – a study on

113 patients with low-risk MDS

J. Cermak1, A. Jonasova2, J. Vondrakova3, L. Cervinek4,

P. Belohlavkova5, D. Sponerova1, L. Novakova1, L. Walterova6,

R. Neuwirtova2. 1Department of Clinical Hematology, Institute

of Hematology and Blood Transfusion, 21st Internal Department,

Faculty Hospital, Charles University, Prague, 3Department of

Hematology-Oncology, Faculty Hospital, Palacky University, Olomouc,4Department of Internal Medicine, Faculty Hospital Masaryk

University, Brno, 5Department of Hematology and Internal Medicine,

Faculty Hospital, Charles University, Hradec Kralove, 6Department of

Hematology-Oncology, Regional Hospital, Liberec, Czech Republic

One hundred and thirteen MDS patients with <10% of bone

marrow blasts were treated either with deferiprone (DFPO) or

with deferasirox (DFSX); 48 patients with average initial serum

ferritin level of 2739.5mg/l (range 825–11287mg/l) received DFPO in

a daily dose of 40–90mg/kg, 65 patients with initial serum ferritin

level of 2677.5mg/l (range 780–9923mg/l) received DFSX in a daily

dose of 10–40mg/kg. Median duration of chelation treatment was

10.9 months for DFPO and 13.7 months for DFSX (range 4–36

months). Chelation was effective (maintained or decreased iron

stores) in 73% DFPO treated patients and in 85% DFSX treated

patients with serum ferritin ≤2000mg/l and in 46% of DFPO

patients and 80% DFSX patients with serum ferritin > 2000mg/l.Combination of chelators with rHuEPO (30–40kU/week) enabled

effective chelation in 7 additional patients with serum ferritin

>3000mg/l. Incidence of adverse effects was 62.5% in DFPO group and

56.9% in DFSX group and led to discontinuation of treatment in 29%

of DFPO patients and 6% of DFSX patients. GIT symptoms represented

the most frequent adverse effect (37.5% of DFPO and 33.8% of DFSX

patients) that limited an effective escalation of the daily dose of

the drug. Decreased number of granulocytes was observed in 13%

of DFPO patients and agranulocytosis occurred in 2 DFPO treated

patients (4%), the incidence was similar as in thalassemic patients.

Granulocyte counts restored after cessation of deferiprone treatment

and administration of G-CSF in all but one patient. An increase in

serum creatinine level was observed in 20% of DFSX treated patients.

Deferasirox administered in a daily dose of 15–40mg (depending on

initial serum ferritin level) represents an effective and safe treatment

for iron overloaded patients with MDS. The incidence of adverse