Poster Presentations – 13th International Symposium on Myelodyspastic Syndromes / Leukemia Research 39 S1 (2015) S1–S166 S105 correlated with the type of MDS and the IPSS score, and was tended to increase with disease stage progression. Overall survival and event-free survival differed significantly in accordance with the increasing WT1 transcripts levels by univariate analyses (p=0.038 and P=0.003, respectively). Multivariate analysis also revealed that higher expression of the WT1 and the revised-IPSS category were independent predictive markers for EFS (P=0.031 and P<0.001,respectively). It is noteworthy that the overexpression of WT1 showed a significant effect on EFS in patients with bone marrow blasts less than 5% (P=0.016). In summary, the expression of the WT1 gene can be a useful marker for the diagnosis and risk evaluation of MDS. 210 DECITABINE TREATMENT COULD AMELIORATE PRIMARY IRON- OVERLOAD IN MYELODYSPLASTIC SYNDROME PATIENTS C. Chunkang 1 , S. Gu 1 , Y. Zhao 1 , J. Guo 1 , C. Fei 1 , F. Xv 1 , C. Xiao 1 , X. Li 1 1 Hematology, Shanghai Sixth People’s Hospital, Shanghai, China In order to research how does hypomethylating agents ameliorate iron metabolism in myelodysplastic syndrome (MDS), we performed methylation-specific polymerase chain reaction (MSP), bisulfate genomic sequencing polymerase chain reaction (BSP), quantitative real-time PCR and western blot of hemojuvelin (HJV) and ELISA assay for hepcidin before and after demethylating therapy (decitabine) to determine whether the change of HJV methylation status would have an influence on hepcidin expression. 11 of 22 MDS patients achieved CR or PR according to IWG criteria (50%). HJV mRNA was induced in decitabine responders (P=0.006 comparing pre/post decitabine treatment) but not in non-responders (P=0.121). Similarly, hepcidin serum expression increased from 320.77 ± 34.8 µg/L to 366.77 ± 21.90 µg/L (P=0.012) in responders but did not significantly change in non-responders (P=0.058), while no difference of adjusted serum ferritin (ASF) was found. In conclusion, hypermethylaiton of HJV promoter region could silence the gene expression and demethylating therapy might ameliorate iron-overload through HJV demethylation. 211 SPANISH REGISTRY OF ERYTHROPOIETIC STIMULATING AGENTS STUDY: THE LARGEST RETROSPECTIVE STUDY OF ESAS FOR THE TREATMENT OF ANEMIA IN LOWER RISK MDS PATIENTS M. Diez Campelo 1 , J.I. Lorenzo 1 , L. Benlloch 1 , M. Lopez-Pavia 1 , E. Such 1 , T. Bernal 1 , E. Luño 1 , J. Davila 1 , F. Ramos 1 , M. Calabuig 1 , H. Pomares 1 , B. Gonzalez 1 , B. Merchan 1 , E. Barranco 1 , R. Sancho Tello 1 , M. Callejas 1 , M.J. Requena 1 , M.J. Jimenez 1 , M. Pedreño 1 , A.I. Vicente 1 , A. Medina 1 , A. Campeny 1 , M. Cortes Sansa 1 , C. Pedro 1 , J.F. Falantes 1 , M.J. Arilla 1 , A. Barez 1 , R. Garcia 1 , M.J. Arcos 1 , V. Gomez 1 , C. Muñoz 1 , C. Cervero 1 , J. Casaño 1 , R. de Paz 1 , L. Amigo 1 , A. Insunza 1 , J.A. Muñoz 1 , M.T. Cedena 1 , M. Gomez 1 , P. Font 1 , R. del Campo 1 , C. Fernandez Lago 1 , J.A. Gonzalez Hurtado 1 , M.D. Linares Latorre 1 , A. Mora Casado 1 , M. Vahi 1 , G. F. Sanz 1 , M.C. Cañizo 1 1 Hematology, GESMD, Valencia, Spain Fig. 1. HJV mRNA expression and hepcidin expression before and after decitabine treatment in MDS patients. a. Relative expression of HJV gene in 11 MDS patients (CR & PR group) before and after decitabine treatment (P=0.006). b. Hepcidin expression of 11 MDS patients (CR & PR group) before and after decitabine treatment (P= 0.012). c. Relative expression of HJV gene in 11 MDS patients (HI & PD group) before and after decitabine treatment (P=0.234). d. Hepcidin expression of 11 MDS patients (HI & PD group) before and after decitabine treatment (P= 0.155). Fig. 2. MSP analysis of HJV and BMP2 protein assay in MDS patients who received decitabine treatment. MSP analysis of HJV and BMP2 protein assays in MDS patients who received decitabine treatment (1–6). The outcome of decitabine treatment is: 1.HI, 2.PD, 3-5. CR, 6. PR. Row A: MSP analysis of HJV in MDS patients before decitabine treatment; Row B: MSP analysis of HJV in MDS patients after decitabine treatment. (M: methylated band, U: unmethylated band) As described above, Patient No.2 was PD, while Patient B reached CR. The BMP2 protein expression of Patient No.3 is much higher than Patient No.2. Fig. 3. BSP assay of HJV promoter region in a MDS patient before and after decitabine treatment. a. Before decitabine treatment; b. After decitabine treatment. Red arrow points out the methylation site of CpG islands in HJV gene promoter. As we can see, all the five sites are cytimidine (C) before decitabine treatment, while they changed to thymine (T) after decitabine treatment. The methylation frequency of HJV gene in this MDS patient was reduced after decitabine treatment.