Stockholm, Sweden, June 13 – 16, 2013

cific amplification of the genomic breakpoints and reverse transcription poly - Myeloproliferative neoplasms - Clinical merase chain reaction (PCR).The patient was treated with imatinib and achieved hematologic and cytogenetic remission. Minimal residual disease screening over 3 years with nested PCR failed to detect CEP85L-PDGFRB B1556 mRNA or genomic DNA, confirming a long term molecular remission on ima - RECURRENT CEP85L-PDGFRB FUSION IN A PATIENT WITH A TRANSLO - tinib. In our view, the detection of the exact gene fusion is clinically relevant for CATION T(5;6) AND AN IMATINIB-RESPONSIVE MYELOPROLIFERATIVE effective long term management of these neoplasms as it enables specific fol - NEOPLASM WITH EOSINOPHILIA low up by sensitive molecular analysis. N Winkelmann 1,2* , C Hidalgo-Curtis 1,3, K Waghorn 1,3, J Score 1,3, H Dickin - son 4, A Jack 5, S Ali 6, N Cross 1,3 1Leukaemia Research Group , Wessex Regional Genetics Laboratory, Salis - B1557 bury, United Kingdom, 2Klinik für Innere Medizin II , Universitätsklinikum Jena, CLINICAL SIGNIFICANCE OF IMMATURE PLATELET FRACTION IN BCR- Jena, Germany, 3Faculty of , University of Southampton, Southamp - ABL1-NEGATIVE CHRONIC MYELOPROLIFERATIVE NEOPLASMS ton, 4Cytogenetics Unit, St. James´s University Hospital, 5Haematological N Vazzana 1* , R Spadano 1, S Di Zacomo 2, G Rolandi 3, A Dragani 1 Malignancy Diagnostic Service, St James’s University Hospital, Leeds, 6Hull 1Department of , Centre for Hemophilia and Rare Blood Disorders, Royal Infirmary, Hull Royal Infirmary, Hull, United Kingdom 2Department of , Molecular Biology Unit, 3Department of Transfusion Medicine, Coagulation Unit, Pescara, Italy Background: Fusion genes involving the catalytic domain of tyrosine kinases (TKs) play an important role in the pathogenesis of hematological malignancies Background: Platelet activation plays a pivotal role in the pathogenesis of and solid tumors. In BCR-ABL1 -negative myeloproliferative neoplasms (MPNs) BCR-ABL1-negative myeloproliferative neoplasms (MPN)-associated throm - several different tyrosine kinase fusion events have been described, most com - bosis. Evidence is mounting to support a potential usefulness of immature monly involving the genes encoding the platelet-derived growth factor recep - platelet fraction (IPF) measurement for vascular risk stratification in various tor alpha ( PDGFRA ) or beta ( PDGFRB ). Since the introduction of small mole - thrombotic disorders. cule kinase inhibitors, TK fusions have emerged as prime therapeutic targets. Aims: The aim of this study was to characterize the clinical and laboratory Here, we present the case of a 45 year old male with persistent unexplained determinants of IPF in patients with BCR-ABL1-negative MPN. In addition, we eosinophilia. A bone marrow aspirate and biopsy showed increased cellularity investigated the association between IPF and previous thrombosis. with myeloid expansion and marked eosinophilia without signs of monoclonal - Methods: One-hundred thirty-five patients have been studied. Sixty-one ity. Cytogenetic analysis on a bone marrow aspirate revealed a patients (45.2%) had ET, 25 (18.5%) PV, and 21 (15.6%) MPN-U. Among the 46,XY,t(5;6)(q3?3;q2?2). 28 (20.7%) patients with myelofibrosis, 25 patients had PMF, while 3 patients Aims: In this patient our objective was to investigate the underlying fusion gene had post-TE or post–PV myelofibrosis. Forty-eight patients had a history of of his translocation t(5;6) and if possible use it as a molecular marker during previous thrombotic event, including arterial thrombosis (n = 31), venous throm - treatment. bosis (n = 14), or both (n = 3) events. Complete blood counts, including the Methods: Break apart FISH using previously described in house probes measurement of IPF were performed in whole blood by the fully automated demonstrated that PDGFRB at 5q33 was disrupted but ETV6-PDGFRB, the hematology analyzer XE-2100 (Sysmex). most common fusion involving this gene, was not detected by RT-PCR.Stan - Results: In patients on cytoreductive but not in untreated patients, dard Gold Taq Polymerase based PCRs were performed on cDNA and gDNA IPF% was significantly higher in those with previous thrombosis than in non - extracted from peripheral blood leukocytes. Sanger Sequencing was performed thrombotic patients [2.4 (1.7-3.4) vs. 3.3 (2.4-5.1) %, P=0.011]. Similarly, in on cDNA and gDNA in forward and reverse. Peripheral blood samples were patients aged ≥ 60 years but not in younger patients, IPF% was significantly received from this patient at 7 different time points before and after treatment. higher in those with previous thrombosis than in nonthrombotic patients [2.6 (1.8 -3.7) vs. 3.6 (2.4 - 5.2)]. In the entire population, a significant inverse correla - tion has been observed between platelet count and IPF% (Rho = - 0.23, P=0.008). In addition, in non-PV patients, IPF% was not significantly different between JAK2 V617F positive vs. negative patients. Multivariate logistic regres - sion showed that only male gender (odds ratio, 3.3; 95% CI, 1.4 to 8.0; P=0.007) and the upper tertile of IPF% (odds ratio, 3.7; 95% CI, 1.2 to 10.7; P=0.018) are independently associated with a history of previous thrombosis, after adjust - ing for age, hematocrit, white blood cell count, platelet count, cardiovascular risk factors, underlining diagnosis, JAK2 mutational status and cytoreductive ther - apy. Summary / Conclusion: We found that increased platelet turnover, as reflect - ed by high IPF%, is associated with a history of thrombotic events in patients with MPN. In addition, our data support the hypothesis that current antithrom - botic therapy might not specifically address this mechanism of thrombogene - sis. New prospective studies are warranted to evaluate the usefulness of incor - porating IPF% in risk stratification models to better identify patients at increased risk for thrombotic complications and/or treatment failure.

B1558 DISEASE CHARACTERISTICS AND PERIPHERAL BLOOD CD34+ CELLS IN IDIOPATHIC MYELOFIBROSIS S Improta 1, M Villa 1* , A Gagliardi 1, C Tommasino 2, G Fossati 2, L Mastrullo 1 1U.O.C. Ematologia, 2U.O.C. Patologia Clinica, P.O. San Gennaro ASL Napoli Results: At the time of initial analysis no similar translocation had been report - 1 Centro, Napoli, Italy ed and therefore the diagnostic investigations suggested the likely presence of a novel fusion involving PDGFRB and an unknown partner gene on chromo - Background: Idiopathic Myelofibrosis (IMF) is chronic myeloproliferative neo - some 6. Initial attempts at RACE-PCR were unsuccessful and hampered by lim - plasm characterized by constitutive mobilization of hematopoietic stem cells ited availability of suitable pre-treatment material. Subsequently, a novel (HSC) and progenitor cells (HPC) into the peripheral blood (PB). The interac - C6orf204 -PDGFRB (now known as CEP85L-PDGFRB ) fusion was reported in tion between the chemokine CXCL12 and its receptor CXCR4 plays a pivotal a patient with T-cell acute lymphoblastic leukemia (T-ALL). Thus, we hypothe - role in determining the trafficking of CD34+ cells between the bone marrow (BM) sized that our patient may harbour the same or similar genetic defect. For the and the PB. RT-PCR, a product was amplified from the patient with the t(5;6) but not con - Aims: IMF is associated with downregulation of CXCR4 by CD34+ cells due trols which upon sequencing revealed an in frame cDNA fusion between exon to epigenetic events. Altered gene expression was corroborated by the detec - 11 of CEP85L and exon 12 of PDGFRB. For gDNA, Sequencing showed the tion of abnormally high CD9 or CD164, and low CXCR4, membrane protein genomic fusion was between intron 12 of CEP85L and exonic sequence of expression in IMF CD34+ cells. Moreover, endothelial precursor cells PDGFRB exon 11 (Figure 1) .Five months after the start of imatinib treatment (CD34+/CD133+) are increased in the blood of a subset of patients with IMF, neither the genomic nor the mRNA fusion was detectable by nested PCR or RT- and peripheral endothelial cells bear the same molecular markers as PCR. CEP85L-PDGFRB remained undetectable in all subsequent samples hematopoietic cells, suggesting a primary role of pathological endothelial cells with a follow up of over 3 years. Imatinib was continued without interruption dur - in this disease. ing this time The sensitivity of the PCR assay to detect the CEP85L-PDGFRB - Methods: We evaluated, by flow cytometry, the number of CD34 positive cells fusion gene was 10 -4 . in peripheral blood and the expression of CXCR4, CD9, CD117 and CD133 on Summary / Conclusion: Here, we report a recurrent CEP85L-PDGFRB fusion these cells. In our institution we are following 31 patients affected by IMF, in a patient with eosinophilia and an MPN. The fusion was confirmed by spe - according to WHO criteria (M: 18, F: 13; median age: 57 years, range: 48-68

haematologica | 2013; 98(s1) | 619 18 th Congress of the European Hematology Association years). Hôpital Haut-Lévêque, Bordeaux, 3Shire, Boulogne-Billancourt, France, 4Shire Results: In all patients, at diagnosis, we found a high count of CD34+ cells in Pharmaceuticals Ltd, Basingstoke, United Kingdom, 5Centre d’Investigation PB (greater than 15x10 6/l; median: 2, 4x10 6/l, range: 1, 8-3,2x10 6/l) compared Clinique, Hôpital Saint Louis et Université Paris 7, Paris, France with normal controls and other Philadelphia-negative chronic myeloproliferative neoplasms. In all cases CD34+ cells were negative for CXCR4 while express - Background: Anagrelide (ANA) is indicated in the EU for at-risk patients (pts) ing high intensity CD9. About 40% of CD34+ cells expressed CD133, while 20% with essential thrombocythemia (ET) and at least one of: >60 years; platelet expressed CD117 at low intensity. In no case was detected coexpression of count >1000x10 9/L; history of thrombo-hemorrhagic events; in whom prior ther - CD133 and CD117, suggesting a simultaneous presence of two distinct apy (PT) is not sufficiently effective or well tolerated. The Summary of Product hematopoietic progenitors, endothelial progenitors and myeloid progenitors. Characteristics (SPC) recommends starting ANA at 1.0mg/day, in two divided We monitored every 6 months the phenotypic pattern of CD34+ cells, and after doses and maintaining this dose for ≥1 week. Then the dose may be individu - 36-48 months we observed an increase of myeloid precursors ally titrated to achieve the lowest effective dose required to reduce and/or main - (CD34+/CD117+: 45,7%) compared with a reduction of endothelial precursors tain platelet count <600x10 9/L, ideally at 150–400x10 9/L. The dose increment (CD34+/CD133+: 15,3%) in patients who showed clinical and laboratory signs must not exceed 0.5mg/day per week and 2.5mg is the maximum single dose. of disease progression. There is no recommendation on how to transition from PT to ANA. Summary / Conclusion: By comparing these findings with other clinical data, Aims: This observational study (NCT01192347) aimed to identify switch modal - our results seem to confirm that, according to the natural history of disease from ities used when introducing ANA and determine their influence on 6-month an initial stage towards a fibrotic phase (pancytopenia and/or splenomegaly), (mo) outcomes (including efficacy, tolerability and maintenance on ANA at 6 mo) there was a change in PB CD34+ cells. Immunophenotypic profile of PB CD34+ in 44 clinical sites across France. cells is associated in IMF with patients’ clinical characteristics and may have Methods: Pts were enrolled within 1 mo of switching to ANA; up to 1 mo of ret - potential prognostic application. rospective data were collected. As this was a non-interventional study, dosing schedule and follow-up visits were at the investigator’s discretion. Pts were fol - lowed up for 6 mos. All relevant data were collected and recorded from pt B1559 records at the end of the follow-up. THROMBOTIC CEREBRAL EVENTS IN ESSENTIAL THROMBOCYTHEMIA Results: In total, 177 pts were enrolled (safety set n=175), the majority were C Cecchetti 1* , A Aroldi 1, M Riva 1, E Pogliani 1, E Elli 1 female (62%) and aged >60 years (76%). Median age was 70 years. Median 1Hematology division, San Gerardo Hospital, Monza, Italy, Monza, Italy baseline platelet count was 553x10 9/L. Intolerance to therapy (65%) and inef - ficacy (41%) were the most frequent reasons for treatment switch (factors not Background: Patients with Essential Thrombocythemia (ET) are frequently mutually exclusive). ANA starting doses ranged from 0.3–1.5mg/day. The asymptomatic and many remain so. Symptomatic patients tend to present with SPCrecommended starting dose was used most frequently (53%). However thrombotic manifestations in distinct locations. The thromboses are more com - a notable proportion of pts started on 0.5mg/day (41%). The median ANA dose monly arterial than venous. The thrombotic cerebral events (TCE) are rare, also at study end was 1.5mg/day (range 0.3–4.0mg/day). The method of ANA intro - if ET patients may present a wide spectrum of neurologic symptoms (NS) at duction was consistent with the SPC in 76% of pts. Almost all pts switched to onset or during the course of the disease, secondary to microvascular involve - ANA from hydroxycarbamide (93%). Most pts discontinued PT before ANA was ment introduced (66%; Group A). 22% discontinued PT after introduction of ANA Aims: We retrospectively described CTE occurred in 310 patients affected by (Group B; 17% within the first mo [Subgroup B1] and 5% in the subsequent 5 ET referred in our centre from 1990 to 2012 in order to identify clinical and bio - mos [Subgroup B2]). A further 9% had not discontinued PT by the end of the logical features associated to these complications follow-up (Group C) and 5 pts (3%) were determined to have no PT. At the end Methods: We analysed the incidence of TCE; in particular we evaluated the of the follow-up, 85% of pts were still continuing on ANA, Groups: A (82%), B1 prevalence of JAK2V617F mutation in this setting and the possible role of JAK2 (93%), B2 (100%), C (81%). 71% of pts achieved platelet responses, Groups: mutation in the management of CTE A (67%), B1 (83%), B2 (100%), C (56%); 42% full response (<400x10 9/L) and Results: 33/310 patients (10.6%) presented TCE. The average age of TCE 29% partial response (400–600x10 9/L or a reduction of ≥200x10 9/L). The medi - patients was 71 years, with prevalence of female sex (21 vs. 12 patients). Only an final platelet count was 412x10 9/L and the absolute median change from 4 patients were high risk for 2 o more cardiovascular factors (hypertension, dia - baseline was -94.5x10 9/L. 75% of pts who received ANA in line with the SPC betes, obesity, dyslipidemia, smoking, thrombophilia). 54,5% of patients devel - achieved platelet response vs 54% of those not consistent with the SPC. 46% oped CTE before diagnosis of myeloproliferative neoplasm (MPN), with a medi - pts reported adverse drug reactions (ADRs) all described in the SPC. The most an time of 16 months of latent phase of myeloproliferative disease prior to con - frequent were palpitations (13%), headache (11%), diarrhea 6% and asthenia firmed diagnosis of ET. Of the 22 TCE preceding diagnosis, 18 were arterial 6%. 17% pts discontinued ANA due to ADRs (mainly palpitations or headache). events (8 transient ischemic attack, 9 strokes), only 2 events were venous (2 Summary / Conclusion: 85% of pts remained on ANA at the end of the 6-mo ocular thrombosis). 7 patients (21,2%) developed CTE as presentation of dis - follow-up. ANA was introduced using the SPCrecommended dosing sched - ease. All these patients received anti-platelets treatment starting from throm - ule in 76% of pts and 71% achieved platelet responses. Overall, ANA was well botic event. At time of diagnosis, cytoreductive therapy, mainly with hydrox - tolerated and the most frequent adverse events were in line with the SPC. yurea, was started. Therefore, 11 patients (33,3%) developed TCE after diag - Introducing ANA according to the SPC and subsequently withdrawing PT was nosis of ET, despite of anti-platelet and cytoreduction, with 15% of patients associated with the highest platelet response rates. (5/33) with recurrent neurological thrombosis. The blood count at diagnosis For the France Observatoire Xagrid (FOX) investigators. and at time of TCE was similar, in particular no leucocytosis or extreme thom - bocythemia were present. The median value of hematocrit, white blood cell count and platelets at time of CTE were 41%, 7.720/mm3 and 650.000/mm3, B1561 respectively. JAK2V617F mutation was evaluated in 30/33 patients, with sig - RISK OF LIMPHOPROLIFERATIVE NEOPLASMS IN PATIENTS WITH nificant prevalence of JAK2 positive versus negative patients (83,3% vs. CHRONIC MYELOPROLIFERATIVE NEOPLASMS 16,7%). All patients with recurrent CTE were JAK2 mutated. Overall, 91% of S Cancio 1* , G Soler 1, J Miguel Torregrosa 1, E Caparrós 1, M Osma 1, F Ortuño 1, patients (30/33) are alive, with an average follow-up of 57,6 months G Luengo-Gil 1, V Vicente 1, F Ferrer-Marín 1 Summary / Conclusion: we observed that 10,6% of ET patients presented 1Hematology and Clinical , Hospital Universitario Morales Meseguer, CTE, with high prevalence (75,7%) of cerebral events as first presentation at Murcia, Spain time of diagnosis or as first sign of latent disease, for CTE antecedent to diag - nosis. The majority of cerebral events were arterial. Mostly of patientswere Background: Over the last years, two large cohorts of patients have reported females and presented higher prevalence of JAK2V617F mutation. This may that chronic myeloproliferative neoplasms (cMPN) patients have a significant - lead to the fact that, in selected cases of CTE, where there is evidence of lab - ly higher risk of developing lymphoproliferative neoplasm (LPN) compared with oratory signs suggestive of MPN, the detection of the JAK2 V617F mutation pro - the general population [1,14% (22/1915) and 1, 34% (11/820), respectively vides an early diagnosis of MPN. Moreover, a subgroup of patients (33,3%) (Rumi, Haematologica 2011; Vannucchi, Cancer Epidemiol Biomarkers Prev developed CTE also after ET diagnosis and during antiplatelet and cytoreduc - 2009)]. In most cases, diagnosis of LPN was subsequent to the MPN one tive treatment, with high risk of thrombotic CTE recurrence (15%). This seems (91%), and only in 3 cases (9%) the diagnosis of LPN was synchronic or pre - to suggest that in this selected setting, an enhancement of thrombotic prophy - vious to the MPN one. If any genetic susceptibility exists, a random order of laxis could be proposed, for example with association of cytoreduction and onset of the myeloid or lymphoid neoplasm should be expected . Authors of anti-coagulant oral therapy, in presence of JAK2 mutation or other significant these series attribute the low number of cMPN patients with a previous or con - cardiovascular factors or thrombophilia predisposition current LPN due to a more aggressive behavior of lymphoid neoplasms, so these patients would die before cMPN develops. The molecular mechanisms underline the predisposition of cMPN patients to develop a LPN is not known. B1560 Aims: The aim of our study was to evaluate the frequency and time of onset CHARACTERIZATION OF DIFFERENT REGIMENS FOR INTRODUCING of LPN in patients with cMPN in our health area, and to investigate if specific SECOND-LINE ANAGRELIDE: RESULTS FROM A MULTICENTER STUDY genetic marker for predisposition to cMPN may also contribute to the risk of OF 177 PATIENTS IN FRANCE developing LPN. J Rey 1* , J Viallard 2, K Keddad 3, J Smith 4, P Wilde 4, J Kiladjian 5 Methods: All consecutive patients with newly diagnosed of cMPN in our unit 1Onco-Hematology, Paoli-Calmettes, Marseille, 2Service de Médecine Interne, between 2000 and 2011 were included in this study. Among 155 cMPN cases,

620 | haematologica | 2013; 98(s1) Stockholm, Sweden, June 13 – 16, 2013

92 (60%) were diagnosed of essential thrombocitemia (ET), 37 (24%) of B1563 polycitemia vera (PV), 24 (15.5%) of myelofibrosis and 1 (0,5%) of systemic GENETIC ABNORMALITIES ASSOCIATED WITH THROMBOSIS IN mastocytosis (SM) by using WHO criteria. In addition to carry out JAK2V617F, PATIENTS WITH PH-NEGATIVE CHRONIC MYELOPROLIFERATIVE NEO - c-KIT (D816V) mutation analysis for the SM case, we performed mutational PLASMS screening of hotspots: CBL (exons 8 and 9), ASXL1 (exon 12), N/K-RAS (exons O Mishcheniuk 1* , O Kostyukevich 2, I Dmitrenko 1, I Prokopenko 1, V Sholoyko 1, 1 and 2), IDH1/2 (exon 4), TP53 (exons 4-10) and complete coding regions for S Klymenko 1 RUNX1 and TET2. Mutation analysis was made by conventional Sanger 1Hematology and Transplantology, National Research Center for Radiation sequencing. Medicine NAMS of Ukraine, 2Hematology, Center of Prophylactic and Clinical Results: Of the 155 patients included, 4 (2.6%) developed a LPN: in one of Medicine, Kiev, Ukraine them, LPN onset was 2 years before PV whereas in the other 3 patients (2 TE and 1 SM), diagnosis of both neoplasms was simultaneous. All four cases were Background: Given the fact that one of the main causes of death in Ph-neg - men over 60 years. Distribution of LPN cases was as following: 2 cases of ative chronic myeloproliferative neoplasms (CMPN) patients is thrombotic com - CLL, both 0-A stage; 1 case of T-LGLL (TCR gamma/delta+), and 1 case of B- plication, a consistent pattern of genetic abnormalities associated with throm - cutaneous non-Hodgkin lymphoma. This last patient was the only one who bosis might be useful for polycythaemia vera (PV), thrombocythaemia (ET), and required treatment (CHOP-R and radiotherapy), whereas the rest of the patients primary myelofibrosis (PMF) patients risk assessment. remain in therapeutic abstention. In 2 of 3 patients with classic MPN, Aims: The aim of this study was to check whether JAK2 V617F, factor V gene JAK2V617G mutation was detected, whereas the patient with SM presented the G1691A (factor V Lieden, FVL) and G20210A prothrombin gene (PT) mutation c-KIT mutation (D816V). We were not able to detect any additional mutation has the predictive value for thrombosis in patients with Ph-negative CMPN. genes we had evaluated in patients with both myeloid and lymphoid neoplasms. Methods: We examined clinical and molecular genetic parameters of 103 PV, Summary / Conclusion: Accordingly to previous studies, the risk of develop - 43 ET, 36 PMF patients, and 156 persons with transient myeloproliferative reac - ing a LPN in patients with cMPN is greater in men than women and no corre - tions (TMR). lation with mutational status of JAK2 seems to underlie. By contrast, we did not Results: Seven PMF patients (19%), 6 ET patients (14%), 32 PV patients find a higher incidence of LPN between patients with cMPN over the time, since (32%) and 30 TMR individuals (19%) had at least one confirmed arterial and/or in our series, both onset randomly or simultaneously, likely because venous thrombosis. In JAK2 V617F positive PMF patients thrombotic compli - of the inclusion of LPN patients in early stages of the disease. Despite not cation occurred more often than in PMF patients without mutation (6 of 18 vs. described in the two large serious listed above, SM may also be associate 1 of 18 patients, P=0.043). Relative risk of thrombosis in JAK2 V617F positive rarely to LPN. Although we did not find any additional mutation other than well PMF was 6.0 (95% CI=1.01-44.7; p˂0.05). Episodes of atypical thrombotic known JAK2 V617F in classical MPN and c- KIT (D816V) in SM, we cannot complications manifested in 13 (28%) unselected Ph-negative CMPN patients exclude that different genes other than JAK2 V617F might favor the genetic with thrombotic complication, 2 of them (4%) had both typical and atypical instability predisposing to both LPN and cMPN or that a mutator phenotype thrombosis, all of these patients were JAK2 V617F positive. The FVL allele has exists in these cMPN patients. been detected in 3.3% of Ph-negative CMPN patients (6 of 182 cases) and in 7.7% (12 of 156) of TMR persons. The difference between groups was on the border of statistical significance (P=0.07). The FVL mutation was detected in 3 B1562 of 38 Ph-negative CMPN patients with thrombotic event (7.9%) and in 3 patients SERIAL ANALYSIS OF GENOMIC ABERRATIONS IN DIFFERENT PHASE without thrombosis (2.9%, P=0.2). The separate analysis shown that the preva - OF PATIENTS WITH MYELOPROLIFERATIVE AND lence of FVL mutation was higher in PMF patients with thrombosis compared MYELODYSPLASTIC/MYELOPROLIFERATIVE NEOPLASMS to those without thrombotic episode (2 of 7 vs. 0 of 29 patients, P=0.03). The C Hahm 1,2, Y Mun 3, C Seong 3, W Chung 1, J Huh 1* prevalence of FVL mutation in PV patients did not differ between groups of 1Department of Laboratory Medicine, Ewha Womans University School of Med - patients with and without thrombosis. The PT mutation has been detected in 3 icine, 2Department of Laboratory Medicine, Eone Reference Laboratory, (2.1%) of 142 Ph-negative CMPN patients. The prevalence of PT mutation was 3Department of , Ewha Womans University School of Medi - higher in ET patients with thrombosis compared to patients without this com - cine, Seoul, Korea, Republic Of plication (2 of 6 vs. 0 of 37, P=0.02). Transient myeloproliferative reactions patients with thrombotic episodes demonstrated a higher prevalence of FVL Background: Myeloproliferative neoplasm (MPN) and myelodysplastic/myelo - mutation than those without thrombosis (6 of 30 vs. 6 of 126, p˂0.05). The FVL proliferative neoplasm (MDS/MPN) may transform into secondary myelofibro - mutation in TMR patients was associated with a 4.2-fold (95%, CI=1.46-12.06; sis (MF), and evolve into acute myeloid leukemia (AML), possibly preceded by p˂0.05) increase of probability of having any thrombosis. Relative risk of venous a myelodysplastic phase. Genetic mechanisms underlying disease progres - thrombosis was 5.6 (95% CI =1.79-17.28; p˂0.05) in carriers of FVL allele. The sion remain to be cleared, while chromosomal aberrations are relatively rare in PT mutation has been identified in 8 (5.1%) individuals with TMR. The preva - the chronic phase of MPN. lence of mutant allele was higher in TMR patients with thrombosis compared Aims: The purpose of this study was to identify serial genomic aberrations that to those without the thrombotic event (4 of 30 vs. 4 of 126, P=0.027). It was are associated with disease progression in MPN and MDS/MPN, using whole found that individuals with PT mutation demonstrated a 4.1-fold (95% CI=1.09- genome single nucleotide polymorphism based array (SNP-A), which can detect 15.33, P<0.05) greater probability of having thrombosis. All FVL and PT muta - cryptic aberrations or copy neutral loss of heterozygosity (CN-LOH). tions detected in our study were heterozygous. Methods: We investigated serial genomic changes of MPN and MDS/MPN Summary / Conclusion: Our data confirm that JAK2 V617F mutation is the patients in different phases of disease. The study group included 10 MPN and predictor of thrombotic complication in Ph-negative CMPN and support the 4 MDS/MPN patients (7, polycythemia vera (PV); 2, essential thrombocythemia assumption that FVL and PT heterozygous mutation might further increase the (ET); 1, primary myelofibrosis; 2, MDS/MPN-unclassifiable; 1, chronic myelomono - risk of thrombosis in PMF and ET patients respectively. cytic leukemia; 1, atypical CML). Median follow up time was 78 months (range 8-154). Whole genome SNP-A (SNP 6.0, Affymetrix, CA) based karyotyping was performed according to manufacturer’s instruction. B1564 Results: Four patients (1 PV, 1 ET, 2 MDS/MPN-U) progressed to secondary HEMOGLOBINOPATHIA YPSILANTI – A RARE, BUT IMPORTANT DIFFER - MF or AML. All PV patients except one developing to MF showed CN-LOH of ENTIAL DIAGNOSIS TO POLYCYTHEMIA VERA chromosome 9p at diagnosis and during follow up. One patient with PV devel - M Nygaard 1,1* , J Petersen 2, O Bjerrum 1 oping to MF showed serial genomic changes by SNP-A; CN-LOH of 9p shown 1Dept Hematology , Rigshospitalet, Copenhagen , 2Dept Hematology , Herlev at diagnosis changed to gain (copy number 3) of 9p after 18 months and then Hospital, Herlev, Denmark additional lesions such as 1q gain and 6p deletion were accompanied at 100 months after diagnosis. Moreover, the copy number of 9p increased to four. One Background: Hemoglobin Ypsilanti is a rare high oxygen affinity hemoglobin ET patient developing to MF showed CN-LOH of 9p, 6q deletion and CN-LOH variant involving an amino acid substitution and is inherited autosomally dom - of 14q at diagnosis and the same genomic changes showed during 106 months inant. Due to the high oxygen affinity of hemoglobin Ypsilanti, hypoxia arises in follow up. In the other ET patient without disease progression, any genomic the tissues, and secondary erythrocytosis develops. Before the inclusion of aberrations were not found during 84 months. One MDS/MPN, unclassifiable JAK2/exon 12 mutations in the diagnostic criteria, a patient with erythrocytosis patient demonstrated chromosome 8 gain, 13q deletion and CN-LOH of 20q at could be difficult to diagnose properly with polycythemia vera (PV). The JAK2 diagnosis and throughout follow-up and he progressed to AML at 14 months mutation is present in more than 95% of patients with PV, and in the rest, a after diagnosis. During blastic phase, the same previous aberrations were mutation in exon 12 is almost always found. The number of patients with non- detected and additional lesions such as 5q and 17p deletions were observed. clonal PV is therefore very low, but it is very important to obtain a correct diag - The other MDS/MPN, unclassifiable patient developing to MF showed normal nosis due to correct patient information and treatment strategy. result by SNP-A during 94 months. Aims: To highlight the importance of proper differential diagnostics in PV. Summary / Conclusion: This study suggests that disease progression from Methods: We present a case report of a mother and daughter who were ini - chronic phase to secondary MF or AML is associated with genomic changes that tially diagnosed with PV. can be identified by SNP-A. Results: In 2003, the mother (age 51 years) was referred to the hematologi - cal department due to a high hemoglobin (B-Hgb) level diagnosed during admis - sion for a suspected transient cerebral ischemic attack. In 2007, her daughter (age 24 years) was also referred due to a high B-Hgb level and a three week

haematologica | 2013; 98(s1) | 621 18 th Congress of the European Hematology Association history of headache. Neither had leukocytosis, thrombocytosis or hepato- therapy; its negative impact on quality of life is significant. The ulcers occur spleno-megaly. Chest X-rays and peripheral blood smears were normal. Bone more frequently in patients with CVI and JAK-2 mutation; however their preva - marrow biopsies showed hypercellularity and in the case of the mother, the lence in patients without CVI is not negligible. The role of HU in development bone marrow examinations were interpreted in concordance with a myelopro - of LU is substantial and the discontinuation of HU therapy is required in most liferative disorder. S-EPO was within normal range for both. EPO-stimulated patients. growth of erythroid colonies were normal in both patients. The JAK2/ and exon 12 analyses showed wild-type in both mother and daughter when these analy - ses became available. The mother later developed pulmonary embolism after B1566 immobilization following . Both were treated with venesection regular - THE ROLE OF HISTOLOGICAL FEATURES IN PATIENTS WITH ly according to guidelines for PV to a hct < 45 %, and being comfortable with POLICYTHAEMIA VERA this. The mother also received anticoagulant therapy, first as acetylsalicylate R Latagliata 1* , M Breccia 1, I Carmosino 1, F Vozella 1, A Romano 1, C Montagna 1, and later warfarin after the pulmonary embolism. In 2012, a hemoglobin analy - A Salaroli 1, A Serrao 1, G Loglisci 1, M Mancini 1, A Tafuri 1, G Alimena 1 sis by β-globin gene sequencing was performed and both were heterozygous 1Cellular Biotechnologies and Hematology, UNIVERSITÀ “LA SAPIENZA”, for the mutation CD 99 G>T (Asp99Tyr), and therefore a carrier of the Hgb-vari - Roma, Italy ant Ypsilanti. The regular venesection was stopped and B-Hgb allowed to increase within normal. The daughter has given birth twice to apparently nor - Background: According to WHO revised classification, bone marrow biopsy mal offsprings. is no more needed in the diagnostic process of Policythaemia Vera (PV). How - Summary / Conclusion: The pursuit of a correct diagnosis is important - espe - ever, it was widely used in the past years and is still performed in many PV cially today where molecular biology offers the possibility of accurate diagno - patients at baseline. sis. Erythrocytosis due to high oxygen affinity hemoglobin is usually well toler - Aims: At present, very few data exist on the correlation between histological ated in younger patients, but in elderly patients the risk of thrombosis is features and patient follow-up. increased. Standard venesection treatment with high oxygen affinity hemoglo - Methods: To address this issue, we revised retrospectively the histological binopathies may not be rational, since the high oxygen affinity causes tissue features at baseline of 195 patients with PV [M/F 100/95, median age 58.4 yrs hypoxia, if there is no compensatory erythrocytosis. Contrary to PV, the risk of (IQR 48.1 – 67.2), median Hb 18.4 g/dl (IQR 17.1 – 20.1), median Ht 56.6% transformation to myelofibrosis or leukemia is not present, which has a great (IQR 53.0 – 61.8), median WBC 10.4 x 10 9/l (IQR 8.4 – 14.0), median PLTs 505 impact psychologically. Furthermore, cytoreductive therapy with e.g. hydrox - x 10 9/l (IQR 346 – 717)] observed at our Institution from 1/1982 to 12/2010. yurea or interferon- α is not indicated and may cause side effects, affecting Results: The following histological features were considered: bone marrow quality of life negatively and may imply long-term complications. It is important fibrosis [grade 0 in 128 patients (65.6%) and grade > 0 in 67 patients (34.4%)], to suspect high oxygen affinity hemoglobinopathy when there is a family his - number of megakaryocytes [normal in 34 patients (17.4%) and increased with tory of erythrocytosis and/or when young persons are affected; especially when or without clusters in 161 patients (82.6%)], marrow cellularity [normal or no apparent cause or clonal marker is identified. The finding of high oxygen increased with erythroid hyperplasia in 152 patients (77.9%) and increased affinity hemoglobin is important, for patient information and optimal treatment with granulo-megakaryocytic hyperplasia in 43 patients (22.1%)], lymphoid strategy. reactive infiltration [absent in 165 patients (84.6%) and present in 30 patients (15.4%)]. The different histological features at baseline were compared with thrombotic episodes during follow-up [reported in 45/195 patients (23.1%)], B1565 evolution in myelofibrotic phase (MP) [21/195 patients (10.8%)], evolution in THE PREVALENCE OF LEG ULCERS IN PATIENTS WITH MYELOPROLIF - blastic phase [15/195 patients (7.7%)] and overall survival. Marrow cellularity, ERATIVE DISORDERS TREATED WITH HYDROXYUREA number of megakaryocytes and lymphoid infiltration did not show any signifi - H Poul 1* , P Kessler 1, E Drnkova 2, I Zemanova 2 cant prognostic role: on the contrary, the presence of marrow fibrosis > grade 1Department of hematology and transfusion medicine, 2Department of angiol - 0 was associated with an increased occurrence of thrombotic episodes during ogy, Hospital Pelhřimov, Pelhřimov, Czech Republic follow-up (P=0.020) and an increased rate of evolution in MP (P=0.040). Summary / Conclusion: In conclusion, while the histological marrow evalua - Background: Leg ulcers (LU) represent a common adverse side effect of long- tion is not required for the diagnosis of PV, the recognition of fibrosis seems to term hydroxyurea (HU) therapy, which is often neglected. have a role in the prognostication of adverse events during follow-up and should Aims: To describe the prevalence of LU in patients with myeloproliferative dis - be evaluated at baseline. orders (MPD) treated with HU and association of LU with chronic venous insuf - ficiency (CVI) and presence of JAK-2 (V617F) mutation. Methods: 70 patients aged 39 to 89 years, 33 (47%) women and 37 (53%) men B1567 treated with HU for MPD (polycytemia vera 34 (49%), essential thrombo - COMPARISON OF THE EXPRESSION CANCER-TESTES (CT) ANTIGEN cythemia 14 (20%), primary myelofibrosis 20 (29%), MPD unspecified 2 (2%). PROFILES IN CML AND POLYCYTHEMIA VERA (PV). The average duration of HU therapy was 6.66 years with the average HU dose V Misyurin 1* , A Misyurin 1,2,3, M Sokolova 4, A Krutov 1, L Kesaeva 1, I Soldato - of 2325.8g per patient. JAK-2 (V617F) mutation was examined in 45 (64%) va 1, E Misyurina 1, A Mastchan 5, S Rumyantsev 5, A Rumyantsev 5, A Barysh - patients; positive results were found in 31 (69%) cases. All patients were nikov 2 assessed using the CEAP classification of CVI, including ultrasonography of 1Lab for and Genetic Engineering, Federal Research and lower limb veins. Clinical Centre of Pediatric Hematology, Oncology and , 2N.N. Results: CVI symptoms were found in 51 (73%) patients, 35 (50%) of them had Blokhin Russian Cancer Research Center, 3GeneTechnology LLC, 4Research both lower limbs affected. LU developed in 16 (23%) patients during the peri - Center for Hematology, 5Federal Research and Clinical Centre of Pediatric od of HU treatment. In 11(69%) of them LU was associated with CVI, while in Hematology, Oncology and Immunology, Moscow, Russian Federation 6(38%) cases the ulcer affected the leg without CVI. (1 patient with ulcers of both legs had only one limb with CVI). 19 patients without CVI were followed Background: Since 1951 when W. Dameshek supposed closed interrelation for 19-870 (median 367) weeks and LU developed in 3 (15.8%) of them. 51 within the group of myeloproliferative syndromes many things had happened patients with CVI were followed for 20-933 (median 318) weeks and LU devel - and it turned out that CML differs from other similar diseases by presence of oped in 12 (23.6%) of them; the difference was not significant (P=0.4828). The Ph-chromosome and BCR-ABL gene and that Jak2 V617F and a few other time to LU development was 10 to 547 (mean 233) weeks and 196 to 845 mutations are the molecular hallmarks of PV, ET and IMF. Nevertheless, all (mean 409) weeks in patients with CVI and without CVI, respectively; the dif - these diseases in some features are still quite similar notwithstanding our igno - ference was not significant (P=0.1495). The mean cumulative dose of HU rance about exact causes of this similarity. Cancer-testis (CT) antigens almost administered before LU development was 1848 (55 – 4817) g and 3831 (1635- universally express in different malignancies varying in frequency and intensi - 8648) g in patients with CVI and in patients without CVI, respectively; the dif - ty of expression of particular members of CT antigen family. We have sup - ference was not significant (P=0.1931). The mean duration of HU therapy till posed that CT antigen expression profiles may explain or at least further high - LU development was 257 (10-846) weeks in 7 patients with JAK-2 mutation and light molecular similarity and/or difference between CML and Ph-negative Jak2 313 (197-535) weeks in 3 patients without JAK-2 mutation. The mean cumu - V617F-positive chronic myelogenouse disorder (CMD). lative dose of HU administered was 2238 (57-8648) g and 3243 (1083-7011) Aims: To perform gene expression profiling of a number of CT antigens in g in patients with JAK-2 mutation and in patients without mutation, respective - CML and Jak2 V617F-positive polycythemia vera (PV). ly. 2 (18%) ulcers healed within 18 and 33 weeks during HU therapy. In 15 cas - Methods: In this study we used RQ PCR to analyze and compare mRNA es HU therapy was stopped due to non-healing ulcers and an alternative ther - expression of CT antigens SP17, GAGE1, HAGE, NY-ESO1, MAGE1, PASD1, apy was administered. 14 (93%) ulcers disappeared after discontinuation of HU SCP, SEMG, SLLP1, SPANXA, SSX1 and PRAME in blood cells. Quantitative therapy, 1 patient died shortly after HU suspension. The average time from HU analysis of expression level cancer-testis antigens was carried out relatively discontinuation to healing of LU were 30 (8-73) weeks and 44 (5-121) weeks using the expression of a housekeeping gene ABL as endogenous control to in ulcers associated and non-associated with CVI, respectively; the difference compensate for irregular cell numbers. was not significant (P=0.5035). The average time of healing was 47 (8-121) Results: In blood of CML chronic phase (CP) primary pts (N=36) we observed weeks and 9 (5-13) weeks in patients with JAK-2 mutation and in patients with - HAGE1, SLLP1, SPANXA and PRAME gene expression with frequencies of out JAK-2 mutation, respectively. 61,1% (22/36), 36,1% (13/36), 2, 8% (1/36) and 8, 3% (3/36), respectively. In case Summary / Conclusion: LU is a common complication associated with HU of Jak2 V617F-positive PV pts (N=33) we observed only HAGE1 (63,6%,

622 | haematologica | 2013; 98(s1) Stockholm, Sweden, June 13 – 16, 2013

21/33), SLLP1 (84,8%, 28/33) and PRAME (12,1%, 4/33) gene expression. In is considered as idiopathic (IHES). contrast to CP of CML, expression frequency of SLLP1 and PRAME in PV Aims: to represent our experience in diagnostic and therapy of myeloprolifer - samples was higher (84,8% and 12,1% against 36,1% and 8, 3%). Expression ative diseases (MPD) with eosinophilia and IHES with signs of MPD (myelopro - rate of HAGE gene in both diseases was generally similar (61,1% and 63,6%). liferative variant of IHES). Expression level of genes found in CP of CML did not differ significantly to com - Methods: from 1985 to 2012 years there were 96 patients with HES followed pare with PV. We did not find mRNA expression both in CP of CML and PV: up in our centre. The ratio of male and female was 72:24 and the median age SP17, GAGE1, NY-ESO1, MAGE1, PASD1, SCP1, SEMG and SSX1. In was 38 (17 – 70). Diagnostic steps were directed to verify diseases with primary AP&BC of CML we have observed additional expression of CT22 (14,4%, 1/7), and secondary eosinophilia. MAGE1 (14,4%, 1/7) and GAGE1 (14,4%, 1/7) (Fig.1). For the proof of the clonal MPD existence standard cytogenetic study and, from Summary / Conclusion: Our data suggest obvious similarity of CT expression the 2003 year, the polymerase chain reaction (PCR) with FIP1L1-PDGFRA and profiles of CP of CML and Jak2 V617F-positive PV. This finding reflects clini - ETV6-PDGFRB primers were applied. cal relation being observed among these two leukemias. Both CP of CML and Results: reactive eosinophilia was proved in 16% (14/96) cases: adrenal PV differ in CT expression profile from CML in AP&BC. hemangiopericytoma – 1; multifocal liver damage unknown origin disappeared without cure – 1, allergic reaction – 1; autoimmune diseases – 8, T-cell lym - phomas – 3. It is important to point that the first three cases of eosinophilia could be established postfactum only. The standard cytogenetic study was done in 80 and PCR was done in 61 rest cases of HES. Different chromosomal aber - rations were found by G-banding only in 8% of cases (6/80) whereas PCR could prove clonality in 50% (30/61) of cases with HES. In most cases the FIP1L1-PDGFRA – gene expression was found (28/30). All the rest cases of HES with unestablished cause of eosinophilia were considered as idiopathic. In 19/46 cases there were signs of myeloproliferation and the syndrome was interpreted as myeloproliferative variant of HES. The therapy of clonal MPDs and myeloproliferative variant of HES included schemes with different cytosta - tics, alpha-interferon, hydroxyurea. From the 2003 we began to use imatinib at a dose 100-400 mg o.d. Complete hematologic response (CHR) was received only on imatinib and alpha-interferon. The effectiveness of imatinib was 87% (20/23) in cases PDGFRA- and PDGFRB-positive and 36% (4/11) in cases where PCR was negative or not done. In 14 of 15 cases the complete molec - ular response was received too. At a therapy with alpha-interferon CHR was received in 3 of 7 cases (43%). Neither no hydroxyurea allowed to get any sustained and prolonged result. Summary / Conclusion: About 1/3 of HES cases are represented by clonal MPD to verify which a special methods such as PCR or FISH are required. In B1568 a small percent of HES the association eosinophilia with some diseases can EFFICACY OF PEGYLATED INTERFERON IN MYELO-PROLIFERATIVE be confirmed only after resolution of symptoms. If the target therapy is nonsuf - DISORDERS ficient the possibilities of conservative therapy are limited. For the young S Mc Guckin 1* , J Westwood 1, J Lambert 1, M Scully 1 patients is reasonable to conduct bone marrow transplantation. 1Haematology, University College London hospital, London, United Kingdom

Background: Pegylated Interferon (Pegasus) is an alternative treatment for B1570 patients with myelo-proliferative disorders (MPD), including polycythaemia vera USING MYELOPROLIFERATIVE NEOPLASM SYMPTOM ASSESSMENT (PV) and essential thrombocytosis (ET). FORM (MPN-SAF) TO EVALUATE QUALITY OF LIFE FOR PATIENTS WITH Aims: Review efficacy of Pegylated Interferon in Myeloproliferative Disorders. ESSENTIAL THROMBOCYTHEMIA IN QATAR Methods: We reviewed the use of Pegasus in 13 patients with MPDs (ET=10, M Yassin 1* , H Elayoubi 1, r kamzoul 1, N Eldewik 2 PV=3) at a single institution over a 13 month period; 7/10 ET and 3/3 PV patients 1Hematology/BMT, National centre for cancer care and research, 2qatar med - were JAK 2 V617F positive. ical genetic centre, HMC, Doha, Qatar Results: 10/13 switched from existing including Hydroxcarbamide, Anagrelide and 3x/week Interferon.We recorded: reasons for switching to Pega - Background: Myeloproliferative neoplasms (MPNs), that is, essential throm - sus, starting dose, dose alterations, and any significant side-effects (including bocythemia (ET), polycythemia vera (PV), and myelofibrosis (MF) can lead to derangement in LFTs). Time to response was determined from sequential significant morbidity and mortality among affected patients. Specifically can be platelet/haematocrit readings. 5/13 patients (3 ET and 2 PV) commenced week - predisposed to thrombohemorrhagic events and vascular complications.pro - ly Pegasus as a single therapy, of which 3 (all ET) were treatment naïve. The gressive cytopenias, constitutional symptoms, cachexia , splenomegaly, and remaining 8 patients all had existing therapies withdrawn following initiation of risk of blastic transformation Pegasus, completing the bridging period over 1 to 7 weeks. 1 ET patient Aims: Symptomatic burden in (MPNs) is present in most of MPN patients we remains on Hydroxycarbamide alongside Pegasus. Reasons for switching to sought to use broadly applicable instrument (MPN-SAF) to assess symptoms Pegasus included: ease of weekly administration (n=2), planning to or being in (ET) among populations of Qatar. pregnant (n=4), side-effects of existing therapy (n=3), lower limb ulceration Methods: Using the MF-SAF as a base instrument, we added several key addi - (n=1), non-response to current therapies (n=3). Prior to commencement of tional symptoms previously identified as present in all subtypes of MPNs includ - Pegasus, baseline median platelet count was 387 x10/L (range of 173 – ing headaches, loss of concentration, dizziness, extremity tingling, 1065x10/L). Median platelet counts were 413 (124-889x10 9/L) at 1 month insomnia,sexual problems and mood changes on a 0 (absent) to 10 (worst- and 355 (109-740 x10 9/L) at 3 months after starting therapy. Doses ranged imaginable) scale. The MPN-SAF was administered jointly with the EORTC- from 45µg every 2 weeks to 135µg/week. Within the PV group, there was no QLQ-C30 as the co-validation instrument using prospective cohorts in Qatar change in median haematocrit between baseline (0.50), 1 month (0.48) and 3 (Patients referred to National Centre for Cancer Care and Research). months (0.49). No reported side effects were noted, although 2/13 patients had Results: 123 MPN-SAF surveys were administered (English (45%), Arabic a mean ALT rise of >90IU/L from baseline, following initiation of therapy. (55%) , 78 ET patients were JAK 2 V617F positive and 45patients were JAK2 Summary / Conclusion: Pegylated IFN appears a safe alternative approach V617F negative) an average of 3.6 years from their MPN diagnosis. Participants in MPD therapy with prompt and sustained control of laboratory parameters. were of, age range (22 – 58 years) and gender (52% female) characteristic of disease. Prior hemorrhage (10%) and thrombosis (25%) 78% of patients cur - rently received cytoreductive therapy. 19 items assessed in the MPN-SAF B1569 demonstrated consistently that the most common symptoms were decreased HYPEREOSINOPHILIC SYNDROME: DIAGNOSTICS AND THERAPY. SIN - quality of life (93%), fatigue (84%), insomnia (65%), sad mood (65%), and sex - GLE CENTRE EXPERIENCE uality problems (62%). The least common symptoms (<50% prevalence) were I Nemchenko 1* , A Turkina 1, E Chelysheva 1, G Gusarova 1, A Misiurin 2, L Kesae - fevers (15%), weight loss (10%), abdominal pain (23%), cough (34%), va 2, A Kovrigina 3, E Semenova 3, E Domracheva 4 headache (50%), and bone pain (48%) Interestingly, night sweats (present in 1Scientific Advisory Department of Chemotherapy of Myeloproliferative Disor - 58%) The majority found the MPN-SAF easy to understand (90%) and ders, Hematology Research Centre, 2Molecular Genetics Laboratory “addressed most of my MPN symptoms” (93%). Comparison to EORTC-QLQ- “Genotechnology”, 3Department of Pathohystology, 4Cytogenetical laboratory, C30: Strong correlations existed between individual items represented on both Moscow, Russian Federation the MPN-SAF and the EORTC-QLQC30 including pain, fatigue, appetite and insomnia (all P<0. 001). Additionally key symptomatic elements were highly Background: Hypereosinophilic syndrome (HES) is a group of rare diseases correlated with the EORTC QLQ-C30 functional subscales. Comparison to with persistent eosinophilia >1.5x10/L and organ imparement. It may be оf non - Perceptions: Comparison of the results of the MPN-SAF to enrolling neoplastic or neoplastic origin. If the cause of eosinophilia is not found the HES ’ blinded opinion of patients symptoms (7 assessed - night sweats,

haematologica | 2013; 98(s1) | 623 18 th Congress of the European Hematology Association fevers, fatigue, weight loss,and bone pain) showed excellent correlation with myeloid leukaemia (CML) and myeloproliferative neoplasms (MPNs) have not corresponding patients’ responses (all P<0. 001). Serial MPN-SAF: Pearson been investigated. correlations indicate that most MPN-SAF items are well correlated (r >0. 5, P<. Aims: Using the SEER-Medicare database, fourteen common community 001) upon repeat survey administration. Items characteristic of advanced dis - acquired infections were compared between myeloid malignancy patients AML ease, including weight loss, fever, and cough displayed lower Pearson corre - (n=8,489) and CML (n=3,626) diagnosed 1991-2005; MDS (n=3,072) and MPN lations (r=0. 46, –0. 08, and 0. 38 respectively) (n=2,001) diagnosed 2001-2005) and controls (200,000 for AML/CML and Summary / Conclusion: MPN-SAF is comprehensive and reliable instrument 97,681 for MDS/MPN patients). which is available in multiple languages (including Arabic and English) to eval - Methods: Odds ratios (ORs) and 95% confidence intervals were obtained uate MPN-associated symptoms. The MPN-SAF is recommended as a uniform using polytomous logistical regression with adjustment for gender, age and symptom assessment tool for MPN patient year of diagnosis/selection. Multiple comparisons were considered using Bon - ferroni correction and significant associations analysed over select time peri - NB these data are part of project Sponsored by QNRF NPRP 4-471-31-48. ods. Results: Respiratory tract infections, including bronchitis (ORs 1.20, 1.25), influenza (ORs 1.16, 1.41), pharyngitis (ORs 1.13, 1.22), pneumonia (ORs B1571 1.28, 1.52), sinusitis (ORs 1.23, 1.25) were associated with AML and MDS, PREGNANCY MANAGEMENT AND OUTCOMES IN WOMEN WITH MYELO - respectively. Cystitis (ORs 1.13, 1.26), cellulitis (ORs 1.31, 1.51) and herpes PROLIFERATIVE DISEASES zoster (ORs 1.18, 1.31) were associated with both AML and MDS respective - E Polushkina 1* , R Shmakov 1, M Sokolova 2, N Khoroshko 2 ly while gastroenteritis (OR 1.38) was only associated with MDS. Bronchitis (OR 1Obstetric Physiological Department, Federal State Budget Institution Research 1.21), pneumonia (ORs 1.49), cellulitis (ORs 1.43) and sinusitis (OR 1.19) were Center for , Gynecology and Perinatology Health Ministry of Russia, associated with CML while cellulitis (OR 1.34) was the only infection associat - 2Scientific Advisory Department of myeloproliferative disorders, Federal State ed with MPNs. Claims for most infections occurred 31-72 months preceding Budgetary Institution Hematology Research Center of Health Ministry of Rus - diagnosis although several remained significant >72 months preceding diag - sia, Moscow, Russian Federation nosis. Summary / Conclusion: Stronger associations between community acquired Background: Myeloproliferative diseases (MPD) rarely occur in women of infections and AML/MDS were observed compared to CML/MPNs indicating reproductive age. But in recent years it becomes more often that young women that chronic antigenic stimulation may be important in the malignant transfor - suffer from these diseases. The appearance of new drugs and therapeutic mation of immature blood cells of myeloid lineage. strategies provides good results in survival rate and life prognosis for patients. All this requires special options for management of pregnancy in women with MPD. B1574 Aims: To develop the protocol of preconception planning and pregnancy man - INTERLEUKIN-8 LEVELS IN ESSENTIAL THROMBOCYTHEMIA: CLONAL agement and to evaluate pregnancy outcomes and complications in women OR INFLAMMATORY TRIGGER? with MPD. E Cacciola 1* , A Cipolla 1, E Di Francesco 1, E Seria 1, M Torre 1, R Cacciola 1 Methods: Retrospectively we have analyzed 41pregnancies in 21 women 1Clinical and Molecular Biomedicine, Institute of Haemostasis, Catania, Italy (group 1) who did not receive a special treatment of MPD. The prospective group (group 2) included 46 women who were treated according to our algo - Background: The patients with essential thrombocythemia (ET) overproduce rithm. Our trial included women with main MPD: essential thrombocythemia, proinflammatory cytokines known to promote the hematopoiesis, such as inter - polycythemia vera, primary myelofibrosis. Pregnancy management included leukin-8 (IL-8). The JAK2V617F mutation expressing cells are featured by an examination of blood cell count and hemostasis system twice a month, besides inherited hypersensitivity to cytokine stimulation. this the inherited trombophylia testing, lupus anticoagulant, homocystein lev - Aims: Therefore, we evaluated platelets, red blood cell (RBC), haemoglobin el, antiphospholipid syndrome diagnostics and hematologic examination includ - (Hb) concentration, hematocrit (HCT) and white blood cell (WBC), as myelo - ing trepanobiopsy and JAK2V617F mutation. Besides thorough laboratory proliferative markers, fibrinogen (Fg) and C-reactive protein (CRP), as inflam - examination our protocol of pregnancy planning and management included matory indicators, JAK2V617F mutation and IL-8. cytoreductive therapy, antiaggregants, low-molecular-weight heparin, plasma - Methods: We recruited 50 patients with ET who fulfilled WHO criteria. Their pheresis, vitamins of group B. For the cytoreductive therapy we prescribed mean duration of disease was 10 years (range, 5-20 years). Of 50 ET patients, Interferon alfa which is the safest option in preconception planning and preg - 25 were JAK2V17F mutated (11 males and 14 females, mean age 59 years) nancy management for women with MPD. and 25 were JAK2V617F WT (9 males and 16 females, mean age 58 years). Results: Medical abortion was made in 3 (7,3 %) women, spontaneous mis - All patients were on aspirin. carriages occurred in 65,8% in group 1 without special treatment. First and sec - Results: The JAK2 mutated patients had higher platelets, RBC, Hb, HCT and ond trimester spontaneous abortions prevailed among all the miscarriages – WBC (909±253x109/L, 5.42±7x106/L, 15.2±1.7 g/dl, 46±4%, 11±2.7x109/L) 15 (36,6%) cases, stillbirth occurred in 10 (24,4%) cases. Preterm labor were than JAK2 WT patients (753±129x109/L, 5±7x106/L, 12±2 g/dl, 39±3%, 7± in 14,6 % pregnancies, full-term delivery occurred in 17,1% of cases. Medical 1.5x109/L) (P=0.009, P=0.001, p <.0001, p <.0001, p <.0001), whereas the abortion was made in 2 (4,3%) women of group 2. Spontaneous miscarriages JAK2 WT patients had higher Fg and CRP (414±3 mg/dl and 6±1 mg/L) than occurred only in 4, 5% in the group of women who were treated according to our JAK2 mutated patients (286±37 mg/dl and 0.8±1 mg/L) (p <. 0001 and p protocol (2 pregnancies). Five (10,9%) pregnancies ended preterm, while 37 <.0001, respectively). The JAK2 mutated patients and JAK2 WT patients had (80,4%) labor were fill-term ( P= 0,000001, OR – 0,07; 95% C.I.: 0,018;0,232). elevated IL-8 (274±158 pg/ml and 222±90 pg/ml, respectively). Pregnancy was uncomplicated in 2 (15,4%) and 14 (33,3%) cases in 1 and 2 Summary / Conclusion: On basis of these results it is conceivable that IL-8 groups respectively. The most often pregnancy complications were threaten - may represent the clonal or inflammatory counterpart in mutated and WT ing miscarriage - 10 (76,9%) and 22 (52,4%) cases, – 4 (30,8%) and patients, respectively. Hence, IL-8 may be an unfavourable prognostic index in 13 (31%) in 1 and 2 groups respectively. Placental insufficiency complicated mutated ET patients.r 30,8% pregnancies including intrauterine growth retardation (IUGR) in 2 cas - es in group 1. Although all pregnancies in group 2 were carefully observed and treated 11,2% of them were complicated by placental isufficiency with IUGR in B1575 3 (10,3%) cases. PRIMARY MYELOFIBROSIS – A SURVEY BASED ON THE 20-YEARS’ Summary / Conclusion: Thus pregnancy losses in women suffering from MPD EXPERIENCE OF A SINGLE CENTER occur in 65,8% without special treatment and complications of pregnancy – in V Shuvaev 1* , A Abdulkadyrova 1, V Udaleva 1, R Golovchenko 1, I Zotova 1, I Mar - 84,6% cases. The development of protocol for preconception planning and tynkevich 1, L Martynenko 1, M Ivanova 1, N Cybakova 1, E Petrova 1, M pregnancy management resulted in considerable decrease in miscarriages Kozlovskaya 1, M Fominykh 1, K Abdulkadyrov 1 to 4, 5% ( P= 0,000005, OR – 24,2; 95% C.I.: 4, 8;225,9) and a tendency of lower - 1Russian Research Institute of Hematology and Transfusiology, Saint-Peters - ing of pregnancy complications to 66,7% (P=0,5). burg, Russian Federation

Background: As a result of advances in decryption of molecular mechanisms B1573 of pathogenesis and invention of target drugs, the chronic myeloproliferative COMMON COMMUNITY INFECTIONS AND THEIR ASSOCIATION WITH diseases including primary myelofibrosis (PMF) undergo the renaissance of MYELOID MALIGNANCIES interest nowadays. The information about historical control is needed to assess G Titmarsh 1* , M McMullin 2, C McShane 1, M Clarke 1, E Engels 3, L Anderson 1 potential effect and additional costs of new diagnostic and therapeutic technolo - 1Centre for , Queen’s University, 2CCRCB, Belfast, United King - gies. dom, 3National Cancer Institute, Bethesda, United States Aims: The objective of our study was to review the experience of PMF diag - nostic and treatment in our center for the past twenty years. Background: Antigenic stimulation precedes many haematological malignan - Methods: Our institution has a primary hematological outpatient department cies. Limited evidence suggests that community acquired infections may which works for about a half of Saint-Petersburg city inhabitants – that is about increase risk of acute myeloid leukaemia (AML) and 2.4 million people.We reviewed patients’ charts to obtain information about (MDS). However, associations between community infections and chronic incidence, symptoms, diagnostic test results, treatment options and relation -

624 | haematologica | 2013; 98(s1) Stockholm, Sweden, June 13 – 16, 2013 ship to prognostic factors. Statistical methods included descriptive and Kaplan- value) after starting of induction imatinib therapy. Duration of induction therapy Meyer method with log-rank test for survival comparisons in Statistica 7.0 pack - was at least 3 months Confirmation of complete molecular response was per - age. formed 3-12 months after starting the imatinib therapy. After normalization of eosinophils count and achieving of molecular response maintance therapy was started ( imatinib 100 mg weekly). Confirmatory blood count was performed every 3 months and molecular analysis was performed every 6-12 months. All performed blood count showed normal eosinophils count and nested PCR was negative for the FIP1L1/ PDGFRA gene during maintenance therapy in all patients. Summary / Conclusion: As the maintenance therapy, imatinib 100mg week - ly is effective to sustain the molecular remission in patients with diagnosed CEL.

B1577 A RETROSPECTIVE ANALYSIS ON THE IMPACT OF PROGNSOTIC FAC - TORS IN 362 PATIENTS WITH PRIMARY MYELOFIBROSIS FOLLOWED IN THE LAZIO REGION DURING A 20 YEARS FOLLOW-UP PERIOD S Mecarocci 1* , A Rago 1, R Latagliata 2, A Andriani 2, M Cedrone 2, L Annino 2, A Spadea 2, M Breccia 2, M Montanaro 2, N Villivà 2, S Crescenzi 2, R Porrini 2, E Montefusco 2, M De Muro 2, G Avvisati 2, F Spirito 2, A Bagnato 2, G Alimena 2, G Cimino 1 1Sapienza University, latina, 2On Behalf of the Gruppo Laziale SMPC, Ph1 neg, roma, Italy

Background: Primary myelofibrosis (PMF) is a rare chronic myeloproliferative disorders characterized by a heterogeneous clinical presentation, a relatively Results: Since 1993 to 2012 there were 296 newly diagnosed PMF patients in shortened survival and a propensity to develop acute myeloid leukemia (i.e. our center. This yields stable incidence varied from 0.76 to 1.56 with mean of blast phase). 1.06 new patient per 100 000 inhabitants per year. The age interval was Aims: To correlate the clinic-biologic features at presentation with risk of blast between 16 to 83 years with median of 62 years. The gender ratio was nearly phase evolution, we retrospectively collected the clinical records of 362 patients 2:1 as female:male (192 females and 104 males).The most prevalent symptoms followed in 12 hematologic units of the Latium region and diagnosed between of disease were: splenomegaly (71.4%), constitutional symptoms (fever, night 1981 and 2010. sweats, weight loss) (33.3%), thrombosis 25.4%, portal hypertension (5.1%) Methods: Diagnosis was made according to the criteria accepted at the time with esophageal variceal bleeding (1.6%). The most common lab abnormalities when the patient was diagnosed. Two hundred-nine (65%) patients were males, were leukocytosis (80%), thrombocytosis (69.5%), anemia (39.3%), and throm - 153 females (38%) (M/F= 1.4). Patients mean age was 67 years. Hb < 10 g/dl bocytopenia (10.2%). Bone marrow fibrosis as histological findings as grade 0 was present in 85/274 (31%) of patients. Eigtheen/270 (6.6%) patients pre - (prefibrotic stage) were noted in 26.7%, grade 1 in 20%, grade 2 in 29.2% and sented a WBC count >25 x 10 9/L. The presence of Jak2V617F mutation was grade 3 in 24.2% of patients. Cytogenetic banding analyses were performed in checked in 143 patients. However, only 83 patients had JAK2 V617 mutation 76 cases. Normal karyotype was revealed in 72.4% cases, no mytoses had assessed within 1 year from diagnosis. Among these 83 patients 51 (61%) been obtained in 7.9%. Cytogenetic abnormalities were in 19.7% patients with were JA2V617F-positive. 224 (62%) patients received conventional chemo- trisomy 8 and complex karyotype as most frequent (13.3% of whole clonal therapies. abnormalities for each). JAK2V617F was detected in 50 of 101 (49.7%) exam - Results: At time of analysis, 180 patients were alive, 137 dead and 45 lost to ined patients. In addition MPLW515L mutation was revealed in 1 patient. The follow-up. With a median follow-up of 48.4 months (range: 1-252) the actuari - results of stratification by IPSS/DIPSS+ risk score were the following: low-risk al survival rates at 5, 10 and 20 years were 70%, 35 % and 22%, respectively. 27.0%/25.7%, intermediate-I - 38.4%/35.9%, intermediate-II – 14.3%/26.0%, Forty-two of 362 patient (11.6%) progressed to a blast phase after a median high-risk – 20.3%/12.4%. Patients were treated mainly with hydroxycarbamide time from diagnosis of 92 months (range: 2-250 months) for a cumulative inci - monotherapy or in combination with other drugs in 81.9% cases, interferon was dence rate of 13%, 28% and 28% at 5, 10 and 20 years, respectively. At univari - used in 21.0%, glucocorticoids were used in 11.1% patients. Overall 10-years ate and multivariate analyses only Hb < 10 gr/dl and presence of blasts (>1%) survival of patients was 44.4% with median of 7.6 years. Blastic transformation resulted the only independent prognostic factors significantly affecting the blast occurred in 5.7% patients with median time to blast crisis as of 5.1 years. Over - phase occurrence. JAK2 V617F did not impact on progression to blast phase. all survival highly significantly influenced by risk stratification as IPSS and A score system combining these two factors (score 0= none; score 1= 1 of the DIPSS+. Survival curves according DIPSS+ groups are presented in fig.1. 2 factors; score 2 = both factors) significantly discriminate patients at high, Summary / Conclusion: PMF is one of the most common hematological malig - intermediate and low risk of evolution to blast phase (Figure 1). nancies with reduced life duration. Risk stratification systems had high predic - Summary / Conclusion: In conclusion, our retrospective analysis on a large tive value. Introduction of innovative drugs should be evaluated in comparison series of patients demonstrates that progression to blast phase of PMF patients with historical control. may be predicted by a score system that includes anemia and presence of blasts. These latter are easily and worldwide detectable parameters able to help risk stratification and decision making for PMF patients, an issue that has B1576 became extremely relevant in light of the forthcoming availability of the new tar - 100 MG WEEKLY IMATINIB MAINTENANCE THERAPY IS SUFFICIENT TO geted drugs. SUSTAIN THE MOLECULAR RESPONSE IN PATIENTS WITH CHRONIC EOSINOPHILIC LEUKEMIA W Prejzner 1* , A Leszczynska 1, A Hellmann 1 B1578 1Department of Hematology and Transplantology, Medical University of Gdan - THE ASSOCIATION BETWEEN JAK2 46/1 HAPLOTYPE AND SUSCEPTI - sk, Gdansk, Poland BILITY TO MYELOPROLIFERATIVE NEOPLASMS J Ouyang 1* , X Feng 1, M Zhou 1 Background: CEL is a rare myeloproliferative neoplasm with prominent 1Hematology, the DrumTower Hospital of Nanjing University eosinophilia and presence of a FIP1L1- PDGFRA gene. This fusion gene is Nanjing China, Nanjing, China much more sensitive to imatinib than BCR-ABL gene, and usually a dose of 100 mg daily is sufficient to achieve response to therapy and molecular remission Aims: To investigate wether JAK2 46/1 haplotype was the genetic susceptibil - in most of CEL patient. This imatinib dose is used in induction and mainte - ity gene for MPN patients and the relationship between JAK2V617F and 46/1 nance therapy. haplotype. Aims: Is the imatinib dose of 100mg weekly sufficient to maintain molecular Methods: MALDI-TOF MS was used to analyze two tag SNPs (rs10974944, remission in patinets with CEL? rs12343867) of JAK2 46/1 haplotype in 77 MPN patients with known Methods: 5 patients were diagnosed at the Department of Hematology and JAK2V617F status and 92 healthy persons as control. Transplantology Medical University of Gdansk. All patients are male and diag - Results: The 46/1 haplotype in genotype distribution between the two groups nosed was confirm by the presence of FIP1L1-PDGFRA gene detected by the had a statistically significant difference (P <0.05); Individuals carrying at least PCR. The median age at diagnosis was 47 ( range 39-64) . At the diagnosis one variant allele, compared with the wild-type individuals, had an significant - median number of WBC was 16 G/L ( 10,3-29 G/L) and median eosinophils ly increased risk of MPN by 2.31 times[the OR (95% CI) were 2.31 (1.21-4.39) count was 9,2 G/l ( 5, 36-12,0 G/l). Spleen was enlarged ( palpable below costal and 2.31 (1.2-4.47)];While the MPN risk of people carrying the variant homozy - margin ) in 3 of 5 patients. gotes was increased by 7-9 times[the OR (95% CI) were 7.54 (2.27-24.99) and Results: The normalization of eosinophils was achieved at the 25 days ( mean 8.9 (2.69-29.42)]. Genotype distribution difference between the JAK2 V617F-

haematologica | 2013; 98(s1) | 625 18 th Congress of the European Hematology Association negative group and positive group was statistically significant (P <0.05). The other factors into the DIPSS-plus scoring system was based on single institu - V617F-negative group and control group did not exist a statistically significant tion data, which warrant further validation. difference (P> 0.05),while the distribution between the JAK2 V617F-positive Aims: The aim of this study was to analyze the factors associated to disease group and control group had a significant difference (P <0.05). The test of progression and evaluate whether the new DIPSS-plus prognosis model can peripheral blood cell levels by single factor analysis of variance showed that be validated in our independent dataset. the average blood cell levels of patients with 46/1 haplotype mutation compared with that of wild-type patients, white blood cells increased significantly (11.5608/11.5008 vs. 15.3108/16.3779, P <0.05) while neither hemoglobin nor platelet levels between two groups had a significant difference (P> 0.05). Summary / Conclusion: Our results indicate that the 46/1 haplotype mutation contributes significantly to the occurrence of MPN in populations in Nanjing region of China.

B1579 PRIMARY AUTOIMMUNE MYELOFIBROSIS: REPORT OF TWO CASES N Bru 1* , R Vallansot 1, C Talarn 1, M Cervera 1, R Aguinaco 1, T Gimenez 1, J Do Nascimento 1, L Sisinni 1, L Calvente 1, L Escoda 1 1Haematology, Hospital Universitari Joan XXIII, Tarragona, Spain

Background: Primary Autoimmune Myelofibrosis (PAIMF) is a rare disease characterized by pancytopenia, mild leukoerythroblastosis, isolated serologi - cal immune abnormalities and no splenomegaly. Differential diagnosis include idiopathic myelofibrosis, immune myelofibrosis associated with other disor - ders, particularly sistemic lupus erythematous (SLE) and secondary myelofi - brosis. Aims: To describe clinical features, diagnosis and outcome of two patients with PAIMF at our institution. Results: Case 1. A 53 year old female with no relevant medical history was admitted because of several weeks of progressive asthenia and pancytopenia. Isolated bruising in the legs were present and no splenomegaly was found. The Methods: All patients consecutively diagnosed in our center of MF according haemogram showed leukocytes 3.3 x10 9/L (ANC 1.2x10 9/L), Hb 10.5g/dL, Hct to WHO criteria since 2000 were eligible. Clinical-biological characteristics at 29%, Platelets 42x10 9/L. The immune panel revealed positive anti-nuclear anti - presentation and complications during evolution were registered. Based on bodies 1/40 in a mottled pattern and polyclonal hipergammaglobulinemia. JAK- EUMNET criteria, progression was defined as appearance of constitutional 2(V617F) was negative. Paroxysmal nocturnal hemoglobinuria (PNH) was ruled symptoms; or, decrease of Hb ≥ 20 g/L, transfusion requirement for non-trans - out by flow cytometry. Bone marrow trephine biopsy showed normal morphol - fusion-dependent patients, or increase ≥ 50% of transfusion requirement; or, ogy and distribution of haematological lineages with megakaryocytes precur - substantial increase of spleen size; or leukemic transformation. Association sors surrounded by reticulinic fibrosis. Methylprednisolone at 1 mg/kg/d was ini - between progression and factors at presentation such as grading of bone mar - tiated with a rapid improvement of clinical symptoms, reaching normal haema - row fibrosis, splenomegaly, LDH, Jak2V617F, circulating CD34+ cells in periph - tological values within 4 weeks of treatment. Prednisone was discontinued eral blood (PB), unfavorable cytogenetic (CG), anemia, constitutional symp - after 4 months and the patient remained asymptomatic, with normal toms, DIPPS and DIPPS-plus risk scale were evaluated by Χ2 test. Progres - haemogram until last follow up, 14 months after treatment discontinuation. sion free survival (PFS) and overall survival (OS) analysis was carried out with Case 2. A 76 year old female with a history of hypertension and diabetes melli - the Kaplan-Meier method. The log-rank test and Cox regression were applied tus was admitted because of constitutional symptoms and pancytopenia. At for univariate and multivariate analysis, respectively. physical examination she was pale with no splenomegaly. The haemogram Results: 26 patients were included. Patients´ characteristics at presentation showed leukocytes 2.61x10 9/L (ANC 1x10 9/L with Dhöle bodies and mild leuko - are shown in Table 1. CG analysis were available in 23 cases: standard risk in erythroblastosis), Hb 6g/dL, Hct 19%, Platelets 94x10 9/L. The immune panel 87% of the evaluable cases (normal: n=15; del 20q: n=3; del 13q: n=2) and revealed positive anti-nuclear antibodies 1/40 in a mottled pattern. JAK- unfavorable risk in the other 13% (+8: n= 2; -5: n=1). Almost 80% of the patients 2(V617F) was negative and PNH was ruled out by flow cytometry. Bone mar - were treated with hydroxyurea and 69% (18/26) did not have any complication. row aspiration was dry tap and trephine biopsy showed hypercelularity with iso - The other 8 patients had different complications including new thrombotic (n=2) lated groups of small megakaryocytes and areas of reticulinic fibrosis. Methyl - or hemorrhagic (n=5) events, infections (n=1), other neoplasms (n=1) or pro - prednisolone at 1 mg/kg/d was initiated with rapid improvement of clinical symp - gression (n=5, 2 because of leukemic transformation and the other 3 because toms, reaching normal haematological values within 3 weeks. After 2 months of marrow failure). Two factors associated to disease progression: constitution - treatment was discontinued and the patient remained asymptomatic with nor - al symptoms (P=0.01) and CD34+ cells in PB (174±100/uL vs 79±90/uL, mal haemogram until last follow up 5 months after treatment discontinuation. P=0.08). PFS was 88 months (range 72-103). Due to sample size, we grouped Summary / Conclusion: PAIMF is an infrequent and benign disease that patients in two groups for DIPPS-Plus: Low+Intermediate (Int)-1 vs Int-2+High should be distinguished from other disorders having myelofibrosis. The main Risk. Applied at the time of diagnosis, DIPPS-Plus was predictive of PFS difference with idiopathic myelofibrosis is mild leukoeritroblastosis without (105±4 in low/Int-1 vs 69±13 months in Int-2/high risk, P=0.01). Univariate marked teardrop poikilocytosis in peripheral blood, lack of clustered or atypi - analysis of the weight of each variable in DIPPS-Plus showed that constitution - cal megakaryocites in bone marrow and absence of splenomegaly. PAIMF al symptoms, leukocytosis >25x10 9/L and transfusion requirement are predic - should also be distinguished from myelofibrosis associated with immune dis - tive of shorter PFS. In the multivariate analysis, however, only the presence of orders, mainly SLE. In cases of co-existing SLE or other immune disorders, constitutional symptoms and leukocytosis >25x10 9/L bordered statistical sig - combined immunosuppressive agents are usually necessary. Due to the excel - nificance. OS was 76 months (range 57-96). An association between risk lent response to corticosteroid therapy, empirical treatment should be consid - increases in the DIPPS-Plus model and decrease in OS (91, 86, 74 y 34 months ered to patients with atypical myelofibrosis. for low, int-1, int-2 and high risk respectively) was observed. Summary / Conclusion: Our data suggest that DIPSS-plus prognosis model can classify patients into categories with a different PFS, when applied either B1580 at diagnosis or throughout disease evolution. CLINICAL-BIOLOGICAL CHARACTERISTICS AND RISK FACTORS ASSO - CIATED TO PROGRESSION IN A SERIES OF MYELOFIBROSIS PATIENTS IN A SINGLE CENTER: VALIDATION OF DIPPS-PLUS PROGNOSIS MOD - B1581 EL TWO CASES OF JAK2 V617F-NEGATIVE PV WITH THE N542-Z543DEL S Cancio 1* , J Torregrosa 1, F Ortuño 1, A Jerez 1, G Ortuño 2, Vi Vicente 1, F Fer - MUTATION IN JAK2 12TH EXON. rer-Marín 1 L Galán 1* , J Nieto 1, F De La Fuente Gonzalo 1, P Gradilla 1, F Fernández 1, A Vil - 1Hematology and Clinical Oncology, Hospital Universitario Morales Meseguer, legas 1,2, M Zarzuela 1, E Mandly 1, S Redondo 1, H Saccomani 1, J Vázquez 2Pathology Department, Hospital Universitario Reina Sofía, Murcia, Spain Paganini 1, G Gómez 1, J Mediavilla 1 1Hematology, Hospital Clínico San Carlos, 2on behalf of Spanish Erithropathol - Background: Identification of myelofibrosis (MF) patients with expected short - ogy Group, Madrid, Spain er survival is important for appropriate decision making. For risk stratification, it is recommended to use International Prognostic Scoring System (IPSS) at Background: The discovery of V617F mutation in Janus kinase gene (JAK2) diagnosis and Dynamic-IPSS (DIPSS) anytime during the disease course. revolutionized the classification and diagnosis of BCR/ABL-negative chronic Marked inter-patient variability within IPSS and DIPSS risk groups has been myeloproliferative disorders (BCR/ABL- CMPDs), which includes polycythemia observed, suggesting a potential role of other risk factors. Incorporation of 3 vera (PV), essential thrombocytosis (ET) and primary myelofibrosis (MF). Nev -

626 | haematologica | 2013; 98(s1) Stockholm, Sweden, June 13 – 16, 2013 ertheless, in a great number of cases, these pathologies exist in absence of this Results: Gastrointestinal symptoms (nausea, urge to vomit, abdominal pain, mutation, so it must be other alterations related with their origin and develop - feeling of satiety) were found in 36.99% of patients with CMPD. The highest per - ment. It has been demonstrated that mutations in 12 th exon of JAK2 are centage of gastrointestinal symptoms was observed in patients with IMF involved in the PV development. Until date, 37 different mutations has been (46.15%), while in the other group recorded percentage was significantly less, found in the adjacent region to pseudokinase domain of JAK2 (JH2), all of them but not statistically significant. The presence of various forms of gastrointesti - curses with a PV phenotype similar to that caused by JAK2 V617F mutation. nal bleeding (melena, hematochezia, hematemesis) were observed in less than Aims: Description of two cases of JAK2 V617F-negative PV with a mutation one fifth of patients with CMPD (19.18%). The highest percentage of bleeding (n542-z543del) in JAK2 12 th exon. was originating from the proximal gastrointestinal tract (80.2%), as a result of Methods: We studied 29 patients diagnosed as PV according to the WHO cri - gastric and duodenal ulcers. Only 18.8% of bleedings were from the distal part teria (2008). Hematological data were obtained with a hematologic counter of the gastrointestinal tract. In groups with PV, ET, between 30 and 40% of (Coulter LH750). patients were with ulcer disease that was significantly higher in comparison to those with IMF and MPS (p <0.001). The highest percentage was in patients with PV, and immediately after them are those with ET. Patients with PV had an increased incidence of peptic ulcer disease, caused by the thrombosis of small arteries in the mucous membrane of the stomach and intestines, the effects of gastric juice, which is almost always hyperacid in those patients, basophilia and hyperhistaminemia. Summary / Conclusion: Our investigations have shown an increased inci - dence of gastrointestinal bleedings in patients with true polycythemia, but also a significantly higher incidence of those complications in patients with essen - tial thrombocythemia.

B1583 USE OF RADIO-ACTIVE PHOSPHORUS (P 32) IN THE MANAGEMENT OF POLYCYTHAEMIA RUBRA VERA (PRV) AND PRIMARY THROMBO - CYTHAEMIA (ET) IN EAST KENT UNIVERSITY HOSPITALS NHS TRUST To determinate the presence of the JAK2 V627F mutation a PCR-ARMS was (EKUFHT) UK (01/01/2008 – 31/12/ performed with DNA from whole blood obtained by manual methods (salting S Alhassan 1* , I Dharmasena 1, j lindsay 1, c pocock 1, r senthiraman 1, g shabo 2 out). In some cases in which the mutation was not found, due to clinical evi - 1Haematology, 2Nuclear Mdicine, East Kent University Foundation Hospitals dences (like low erythropoietin levels), we sequenced JAK2 12 th exon, using NHS Trust, Canterbury, United Kingdom the same biological source, but employing an automatic method for the extrac - tion (Biorobot EZ1). Automatic sequencing was performed following the suppli - Background: : As per the British Committee for Standardisation of Haematol - er’s instructions in an ABI PRISM® 3100 Genetic Analizer sistem. ogy (BCSH) guidelines for ET and PRV published in 2010 and 2005 respec - Results: From all cases studied, 23 of them (79.3%) showed the JAK2 V627F tively, P32 therapy is effective in controlling blood counts with few acute side mutation, 2 (6.9%) carries alterations in the 12 th exon of the same gene (in both effects. The usual dose is 150-300MBq, and can be repeated after 3 months. cases, the n542-z543del deletion in heterocygotic state, previously described The use of P32 is recommended as second line therapy in those who are old - by Scott et al. 2007) and in 4 cases (13.8%) any of the mutations studied were er than 70 years who fail to respond to or are intolerant of the first line therapy found. Clinical features of these patients are summarized in the attached table; (hydroxycarbamide). P32, the radioactive isotope of Phosphorous, had long they shown middle age at diagnosis (35-50 years), low erythropoietin (EPO) lev - been successfully used in treatment of myeloproliferative disease, namely PRV els, hemoglobin (Hb), hematocrit (Ht) and erythrocyte count (RBC) values over and ET. It is a pure beta emitting radionuclide with a physical half-life of 14.3 the normal range but no splenomegaly neither thrombotic events. days, given intravenously or orally in aqueous solution. It is actively incorporat - Summary / Conclusion: The 86,2% of patients diagnosed as PV carried muta - ed into the nucleic acids of rapidly proliferating cells, thus suppressing hyper - tions in JAK2 (23 V617F + 2 n542-z543del). Both mutations affect the function proliferative cell lines rather than eradicating them. Early side-effects of tran - of JH2 domain, which loses their regulatory function on the catalytic domain, sient leucopoenia and thrombocytopenia are observed. The late potential for being constitutively active and over activate their related molecular pathways. leukaemogenicity is a risk but its incidence is comparable to that associated with This explains, at least in part, the features of the disease. the chemotherapeutic agents commonly used in management of these condi - So, is not strange that the phenotype caused by both mutations was similar but, tions. (EANM procedure guideline for 32 P phosphate treatment of myeloprolif - in cases with exon12 mutations, the age at diagnosis was lower and the hema - erative diseases; Eur J Nucl Med Mol Imaging (2007)34:1324-27) tological parameters a little higher (with predominant erythrocytosis). These Aims: To audit the efficacy of radio-active phosphorus(P32) in the management data are consistent with those founded in the literature. of PRV and ET and the impact on the haematology out-patient clinic case-load Furthermore, there are four cases where mutations studied were not found, but in EKUFHT. showing a clear PV phenotype (so, it must be caused also by other molecular Methods: Data regarding the total number of episodes, Sex and Age group was alterations in JAK2 or other genes). These facts show that other mutations in collected via the Patient Administration System (PAS) of EKUFH NHS Trust. adition to JAK2 V617F, even if minorities should have a great importance in Data regarding the original diagnosis and outcome was collected from the CMPDs. That might explain phenotypic differences between patients and forms APEX system used in the department of EKHUFHT and from outpa - of the disease (PV, ET, MF) and their implications in the clinical, diagnosis and tient clinic letters. Response was assessed using the following criteria: Com - treatment of the CMPDs must be studied more deeply. plete Response (CR) = HCT < 0.450 + Platelet count <600X10 9/L at 3-6 months post P32 injection + Not on any cytotoxic drugs OR venesection OR Interfer - on-alpha OR Anagrelide + being considered for discharge from clinic; Partial B1582 Response (PR) = reduced dose of cytotoxic therapy OR reduced frequency of PHILADELPHIA CHROMOSOME - NEGATIVE MYELOPROLIFERATIVE venesections OR on Anagrelide/Interferon alpha (alone/in combination).No DISEASES AND GASTROINTESTINAL BLEEDINGS Response (NR) = No change in management plan. E Simonovic 1* , L Macukanovic/Golubovic 2, M Mirjana 1, V Colic 1 Results: Between 01/01/2008 – 31/12/2012 there were 43 episodes (21 1Internal, General Hospital Leskovac, Leskovac, 2Internal, Clinic of Haematol - male/22 female) of usage of radio-active phosphorus (P32) in EKHUFT (23 ogy, Nis, Serbia (53.5%) episodes for treatment of PRV and 20 (46.5%) episodes for treatment of ET). 25 (58.1%) episodes were CR, 8 (18.6%) were PR and 10 (23.3%) Background: Philadelphia chromosome-negative chronic myeloproliferative were NR. This led to 18 episodes of discharge from the haematology out-patient diseases (CMPD) are a group of malignant clonal diseases of hematopoietic clinics (41.9% of the total episodes). stem cells, which are characterized by an overgrowth of one or more blood lines Summary / Conclusion: Radio-active phosphorus (P32) is an effective modal - with normal or nearly normal maturation of those cells in the bone marrow and ity of treatment for those over 70 years of age with a diagnosis of PRV or ET, extramedullary hematopoietic organs. In dependence on the dominating type which reduces/abolishes outpatient clinic attendance; thus contributing to of cells, morphological and clinical characteristics, we include in this group the improved quality of life. following diseases: 1) Polycythemia vera (PV); 2) Agnogenic myeloid metapla - sia with myelofibrosis or idiopathic myelofibrosis (IMF); 3) Essential thrombo - cythaemia (ET); 4) Myeloproliferative diseases that cannot be classified (MPS). Aims: The aim of the paper is to determine what is the incidence of gastroin - testinal bleedings in patients with chronic myeloproliferative disorders. Methods: The investigation included 155 patients of both sexes aged between 17 and 83 years with a diagnosis of CMPD. Patients with a diagnosis of CMPD are divided into four groups. We used methods of clinical examination, labora - tory tests, ultrasound examination of the abdominal organs and esophagogas - troduodenoscopy and rectosigmoidoscopy.

haematologica | 2013; 98(s1) | 627 18 th Congress of the European Hematology Association

B1584 B1585 2 CASES OF IDIOPATHIC HYPEREOSINOPHILIC SYNDROME: A RARE SINGLE-AGENT LENALIDOMIDE IS EFFECTIVE FOR TRANSFUSION DISORDER WITH SEVERE THROMBOHAEMORRHAGIC COMPLICA - INDIPENDENCE IN A PATIENT WITH REFRACTORY ANEMIA WITH RING TIONS SIDEROBLASTS, THROMBOCYTOSIS AND JAK2 (V617F) N Prasannan 1* , A Soultati 1, C Harrison 1, T Yeghen 2, D Radia 1 I Nichele 1* , M Ruggeri 1, F Rodeghiero 1 1Haematology, Guys and St Thomas Hospital, United Kingdom, 2Haematology, 1S. Bortolo Hospital, Vicenza, Italy, Vicenza, Italy Lewisham Hospital NHS Trust, London, United Kingdom Background: Idiopathic hypereosinophilic syndrome (HES) is a rare diagno - Background: Refractory anemia with ring sideroblasts associated with marked sis with a poor prognosis. Management of HES is challenging due to hetero - thrombocytosis (RARS-T) is a rare myelodysplastic/myeloproliferative disorder, geneous clinical presentation. Complications include haemorrhage, thrombo - that has been proposed as a provisonal entity in the 2001 and 2008 WHO clas - sis, multi-organ failure, rapid clinical deterioration and death. Thromboses can sification. JAK2-V617F has been shown in the majority of these patients. The vary from ischemic colitis, sinus thrombosis to DIC and TTP (Ohguchi clinical course of RARS-T could be better than that of RARS and worse than ,2009).There is an urgent need to control the eosinophil count and prevent that of essential thrombocythemia; however, the clinical and therapeutic expe - thrombosis and haemorrhage. Conventional treatment includes steroids, rience with such cases is limited, due to the rarity of this disease. Lenalidomide hydroxycarbamide and interferon-alpha. Several case reports document the is effective in patients with myelodysplastic syndromes with or without the utility of cytotoxic agents such as AraC either alone or in combination. del(5q) cytogenetic abnormality to reach transfusion independence, while no Aims: We describe two cases of HES with eosinophil counts > 100x10 9/L high - efficacy data of Lenalidomide are available in essential thrombocythemia to lighting diagnostic and management challenges. reduce platelet count. The efficacy of single-agent lenalidomide in RARS-T Methods: CASE 1: 60 year old female on rivaroxaban thromboprophylaxis has been recently reported in one report (two patients described). post knee replacement presented with swelling and discolouration of her right Aims: We report here the clinical outcome of Lenalidomide in the treatment of leg. Ultrasound Doppler was negative for a deep vein thrombosis. DVT was one patient with RARS-T and JAK2 V617F. excluded and intramuscular bleed was diagnosed, rivaroxaban was stopped. Methods: A 58-year-old caucasian man was admitted at our hospital in Sep - FBC was Hb 7.6g/dl, WCC 7.5x10 9/L, eosinophils 0.3x10 9/L and platelets tember 2006 for anemia (Hb 9.8 g/dL) and thrombocytosis (platelet count: 1163 265x10 9/L. She was readmitted the following week with exertional x10 9/L). The bone marrow showed approximately 80% cellularity, increased dyspnoea.FBC now Hb 10.5g/dl, WCC 100x109/L (Eo 83x10 9/L). Prednisolone atypical megacaryocytes with often lobulated nuclei and 100mg and heparin anticoagulation commenced for suspected drug- related with 30% ring sideroblasts. Cytogenetic analysis showed a normal karyotype, hypereosinophilic syndrome. BM: mature eosinophilic hyperplasia with normal FISH examination was negative, while PCR revealed the presence of JAK2 maturation. Cytogenetics and FIPILI-PDGFRA status negative. Within 24 hours V617F; a diagnosis of RARS-T associated to JAK2 mutation was made. Due she developed acute onset right hand weakness and CT/MRI imaging suggest - to increase platelets count (1340x10 9/L), hydroxyurea was started after 11 ed transverse sinus thrombosis. Hydroxycarbamide was started, eosinophils months from diagnosis, with worsening of anemia (Hb 9.6 g/dL pre-therapy to remained high. 4 days later, acute confusion occurred. WCC was 134 x10 9/L 8.3 g/dL after three months of cytoreduction). Mild steroid therapy (prednisone (Eo 118x10 9/L). AraC 20mg bid normalised eosinophil counts within a week. 10 mg/day for 16 months) showed a transient efficacy while treatment with However falling GCS and hypoxia developed. Repeat CT confirmed multiple recombinant erytropoietin (30.000 units/weekly) was no successful. Thus, blood cerebral infarcts with small haemorrhages, and anticoagulation discontinued. red cells transfusion treatment was started, after 40 months from diagnosis. 7 days later a myocardial infarction occurred and neutropenic sepsis with acute Because of a high transfusion needs (2 units every months), Lenalidomide 10 eosinophilic lung disease. Multi-organ failure progressed with further acute mg daily for 21 days consecutive was started after 48 months from diagnosis cerebral events leading to signs of brainstem infarction/bleed and the patient and Hydroxyurea was concurrently interrupted. died a few days later. CASE 2: 55 year old male admitted with headache, flu- Results: After 3 cycles of lenalidomide, anemia improved (Hb 9 g/dL) and the like symptoms and bilateral purpuric rash. WCC 16.4x109/L,mild neutrophilia platelet count decreased to 700 x 10 9/L. and anaemia. CT revealed hepatosplenomegaly, mediastinal lymphadenopa - After 6 cycles, the hemoglobin level reached a stable value at > 9 g/dL without thy and adrenal mass. Investigations for vasculitis and infective causes were need of blood red cell transfusion and the platelet count reached the stable val - excluded. The rash progressed WCC 69x10 9/L, Eo 39x10 9/L, BM: reactive ue of 500-600 x 10 9 /L. eosinophilia, no evidence of CML/MPN. Commenced prednisolone for sus - Currently, the patient successfully continues 21 days-cycles of 10 mg lenalido - pected vasculitis. Extension of his rash, ulcers and pneumonia occurred with mide and he maintains the transfusion independence, after a total of 28 cycle WCC 184x10 9/L, Eo69.9x10 9/L. Methylprednisolone and broad spectrum in 2 years of follow-up. No adverse event have been recorded until now. antibiotics were initiated. He developed ascites and coagulopathy. Repeat BM Summary / Conclusion: Therapeutic experiences with RARS-T cases are consistent with HES, commenced Hydroxycarbamide, but continued to deteri - limited. We confirm that Lenalidomide as single agent is efficacy in the treat - orate with confusion, progressive MRSA pneumonia needing ventilation. Inter - ment of our patient with RARS-T and JAK2 V617F, to reach transfusion inde - feron alpha was not effective. Neurological progression with unilateral weak - pendence and to maintain the platelet count within the target threshold after a ness; CT: multiple cerebral infarctions, ground glass shadowing in lungs and long follow-up. Biological and histological analysis to demonstrate the morpho - bronchial lavage revealed eosinophilic infiltrate. AraC 10mg bid controlled the logical and molecular response are being carried out. eosinophil count. Ventilation was required for 7 weeks and on-going Hydroxy - carbamide and interferon alpha. He was discharged for intensive rehabilitation and continues to improve. Results: As above Summary / Conclusion: Our cases illustrate diagnostic and management challenges; rapid disease course reinforces the need to urgently control the counts to minimise the thrombo-haemorrhagic complications. Early use of AraC seems an effective cytoreductive agent in HES with extreme eosinophilia.

628 | haematologica | 2013; 98(s1)