Morphology, Cytogenetics and Survival In myelodysplasia with Del(20q) or Ider(20q): a Multicentric Study

Abstract Abstract 2942 Introduction: In 2004, a variant of del(20q) was described in MDS, i.e. an isochromosome of the long arm of chromosome 20 with interstitial loss of material [ider(20q)]. At the present time, 40 cases of ider(20q) have been reported in hematological malignancies, i.e. twenty-nine in MDS. The reported frequency of ider(20q) compared to del(20q) in MDS and AML, is 0.49% versus 2.49% and 0.26% versus 2.63%, respectively. No specific morphological anomaly was reported for MDS with del(20q) or ider(20q). Del(20q) preferentially involved the erythroid and megakaryocytic precursors and was associated with elliptocytosis. The prognostic significance of del(20q) is generally considered as good with some controversy. Until now, the prognostic significance of ider(20q) remains debated. Aims: The discovery of highly dysplastic neutrophils with neutrophil erythrophagocytosis in a patient with MDS and ider(20q) prompted us to analyze and compare the morphology of blood (PB) and bone marrow (BM) cells in MDS patients with del(20q) or ider(20q). The second aim was to compare clinical features of patients with MDS associated with del(20q) and ider(20q). Materials and methods: 34 patients with MDS displaying del(20q) and/or ider(20q) were included in this retrospective multicentric study. May-Grunwald-Giemsa stained PB and BM smears were analyzed for dysplasia in the erythroid, myeloid and megakaryocytic lineages. All cases were revised independently by at least 2 cytologists from 2 or 3 centers. Cytogenetic analysis was performed at diagnosis on short term BM cultures using conventional techniques, and karyotypes were described according to the International System for Human Cytogenetic Nomenclature (ISCN 2009). When possible (n=20), metaphase and interphase FISH studies were performed on the same suspension as used for conventional cytogenetic analysis to confirm the presence of ider(20q) when cytogenetically suspected, to look for subclones with del(20q), and to establish the percentage of nuclei with ider(20q) and del(20q). All patients were followed from the date of clinical diagnosis to transformation in acute leukemia or death. The presence of autoimmune manifestations and the International Prognostic Scoring System (IPSS) were also registered. Results: For the first time, morphological abnormalities on PB and BM were found sensitive and specific for this recently described cytogenetic entity. Indeed, hypogranulated and vacuolized neutrophils and neutrophil erythrophagocytosis (Figure 1) were present on PB and BM in MDS with ider(20q), with a sensitivity ranging from 65% to 75%. Moreover, the sensitivity and specificity of a morphological score based on the assessment of 9 morphological criteria reached 70% and 90.5%, respectively, in MDS patients with isolated ider(20q). This score included hypogranular and vacuolized neutrophils (in association with non dysplastic neutrophils), hypogranular and vacuolized eosinophils (with sometimes difficult differentiation between neutrophils and eosinophils), neutrophil erythrophagocytosis, elliptocytosis and thrombocytophagocytosis on PB. Hypogranular and vacuolized neutrophils and eosinophils, erythrophagocytosis by neutrophils and deeply lobulated and hyperlobulated megakaryocytes (stag-horn-like) were observed in BM. In the present study, the outcome of patients with del(20q) and with ider(20q) was not statistically different. Indeed, the number of progressions and deaths was slightly higher in the ider(20q) group. However, the frequency of autoimmune manifestations (AIMs) in del(20q) and ider(20q) group were 9.5% (2/21) and 8% (1/13), respectively (p>0.05). The mean IPSS was 0.5 in both groups. The OS (p=0.3744) and PFS (p=0.2497) did not differ statistically in MDS with del(20q) and ider(20q). All deaths were related to the MDS except for one patient with ider(20q) whose dead was from unknown origin. The median OS and PFS were 68 and 65 months, respectively, in the ider(20q) group whereas they were not reached in the del(20q) group. Conclusion: We proposed a morphological score based mainly on hypogranulated and vacuolized neutrophils and neutrophil erythrophagocytosis on PB and BM which can be used to predict ider(20q) in MDS patients. The prognosis seems not to differ between MDS with ider(20q) and del(20q). Disclosures: No relevant conflicts of interest to declare.

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Evaluation of the Intra-LABORATORY Reproducibility of Percentage of Blast Counting In MYELODYSPLASIA

Blood ◽

10.1182/blood.v118.21.4901.4901 ◽

2011 ◽

Vol 118 (21) ◽

pp. 4901-4901

Author(s):

Soraya Wuilleme ◽

Marion Eveillard ◽

Sophie Barillet ◽

Françoise Accard ◽

Hervé Avet-Loiseau ◽

...

Keyword(s):

Bone Marrow ◽

Scoring System ◽

Who Classification ◽

Conflicts Of Interest ◽

Scoring Systems ◽

Poor Quality ◽

International Prognostic Scoring System ◽

Inter Observer Variability ◽

Morphological Criteria ◽

Blast Cells

Abstract Abstract 4901 In the two International Prognostic Scoring Systems of Myelodysplastic Syndrom (MDS), the percentage of BLAST cells in the Bone Marrow is the most important parameter implicated in score: either directly in the IPSS (International Prognostic Scoring System) (1) or indirectly in the WPSS (Who classification-based prognostic scoring system) (2). In these two systems, as in the recommendations of the WHO 2001 (3), the morphological criteria defining blasts cells compared to promyelocyte cells are not specified. In 2005, the IWGM-MDS (International Working Group on Morphology of myelodysplastic syndrome) (4-6) has established morphological criteria defining Blasts cells. Our objective in this study is to evaluate the reproducibility, among five observers of our laboratory, of the counting of the marrow blasts of 73 myelodysplasia. This study was conducted in several stages. 1st step was to test the implementation of the MDS-Foundation (www.mds-foundation.org/virtualmicroscopy) by 5 observers. The 2nd step was to perform correlation test of the selected cells from RAEB (selected and stained in our laboratory conditions), for the 5 observers. Finally Step 3 was to assess the correlation of blast percentage of 73 bone marrow smears from MDS patients, selected due to the presence of an excess of blast cells in the first reading on the bone aspiration. Our results show that the correlation on counting blasts is generally satisfactory (percentage agreement: Test 1 = 86% and test 2 = 94%), while the concordance on the counting blasts of bone marow smears of 73 MDS patients and concordance on the WHO classification seems less satisfactory (agreement 3/5 observers. = 95% but agreed to 4–5/5 observers = 64%). These results can be explained partly by the inter-observer variability and by the variability of some parameters specific to smear marrow (poor quality smears, the staining and/or poverty of smears). In conclusion, the evaluation of the blasts in the MDS must be achieved: (i) at least 500 cells counted (ii) by at least two different observers (iii) by a third observer in discordant case. Despite these recommendations, the assessment of the blasts in myelodysplasia is difficult to achieve in many cases. Disclosures: No relevant conflicts of interest to declare.

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Cytogenetic Analysis in Patients with Newly Diagnosed Myelodysplastic Syndromes in Southern Italy

Blood ◽

10.1182/blood.v120.21.50623.50623 ◽

2012 ◽

Vol 120 (21) ◽

pp. 50623-50623

Author(s):

Esther Natalie Oliva ◽

Francesco Albano ◽

Nicola Di Renzo ◽

Vincenzo Pavone ◽

Stefano Molica ◽

...

Keyword(s):

Bone Marrow ◽

Myelodysplastic Syndromes ◽

Cytogenetic Analysis ◽

Southern Italy ◽

Conflicts Of Interest ◽

Refractory Anemia ◽

Newly Diagnosed ◽

Conventional Cytogenetic Analysis ◽

Conventional Analysis ◽

Italian Regions

Abstract Abstract 50623 Background: Bone marrow karyotype in myelodysplastic syndromes (MDS) is essential to define the prognosis and to guide treatment decisions, including targeted therapies. Due to the lack of an extensive national MDS registry in Italy, epidemiological data on MDS, including cytogenetics, throughout the territory is unknown. Objective: We evaluated the incidence of cytogenetic abnormalities amongst newly diagnosed MDS patients in in 2 southern Italian regions (Calabria and Puglia). Methods: A pilot project, denominated ANDROMEDA (ANalysis of cytOgenetics alteRatiOn in the MyEloDysplAstic syndromes) was developed in 17 centers to offer a service of conventional cytogenetic analysis for all consecutive patients undergoing diagnostic evaluation for cytopenia and suspected MDS between January 1 and December 31, 2011. The study conformed to the ethical standards set out in the Declaration of Helsinki and was approved by institutional review boards at each participating center. Patients were required to provide their written informed consent. Clinical characteristics of patients and bone marrow morphology, iron staining and histology were registered. Bone marrow samples were centralized for standard cytogenetic studies and fluorescence in situ hybridization to two dedicated genetics laboratories (one for each region), blind to patients' data. Results: Two hundred and thirty-five patients were evaluated and MDS diagnosis was confirmed in 220 cases (88. 3%), according to WHO criteria. The overall incidence of clonal chromosome abnormalities detected by conventional analysis was 36. 9%. Single abnormalities included +8 (13 cases, 5. 8%), del(5q) (12 cases, 5. 4%), –Y (11 cases, 5. 0%) and del(7)/-7 (4 cases, 1. 8%). Complex karyotypes were detected in 18 (8. 1%) cases. Among all cases only 10 (4. 5%) bone marrow samples were not evaluable for cytogenetic analysis. FISH revealed additional abnormalities not identified by conventional analysis only in 3 (1. 3%) out of 72 cases. Patients were classified in WHO subtypes: 39. 2% refractory cytopenia with unilineage dysplasia (RCUD), 1. 5% refractory anemia with ring sideroblasts (RARS), 32. 5% refractory cytopenia with multilineage (RCMD), 10. 8% refractory anemia with excess of blast-1 (RAEB-1), 9. 3% refractory anemia with excess of blast-2 (RAEB-2), 4. 1% MDS with deletion 5q (MDS 5q-) and 2. 6% MDS unclassifiable (MDS-U). Conclusions: These preliminary results demonstrate that the incidence of abnormal karyotype patterns and WHO subgroups in MDS patients in Southern Italy is comparable with that described in other geographical areas. It is confirmed that conventional cytogenetic analysis is a standard in the diagnostic workup of MDS of patients with a suspected myeloid malignancy in order to identify primary abnormalities and prognostic models. Disclosures: No relevant conflicts of interest to declare.

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Diagnostic and Prognostic Significance Of Isolated Trisomy 2 In Hematopoietic Malignancies

Blood ◽

10.1182/blood.v122.21.4970.4970 ◽

2013 ◽

Vol 122 (21) ◽

pp. 4970-4970

Author(s):

Umut Aypar ◽

Lindsey Waltman ◽

Kaaren Reichard ◽

Daniel L. Van Dyke

Keyword(s):

Bone Marrow ◽

Conflicts Of Interest ◽

Prognostic Significance ◽

Refractory Anemia ◽

Related Phenomenon ◽

Diagnostic Features ◽

Hematopoietic Malignancies ◽

Age Related ◽

Morphological Criteria ◽

Bone Marrow Blasts

Abstract Isolated trisomy 2 in hematopoietic malignancies is rare, having only been reported in eight cases in the literature. Of these cases, the majority are older male patients (7/8) ranging in age from 64 to 84 years. The underlying hematologic malignancies include: myelodysplastic syndrome (MDS), refractory anemia (RA) subtype; MDS, RA with excess blasts (RAEB-II) subtype; MDS, RAEB in transformation (RAEB-t); chronic myelomonocytic leukemia (CMML-t) in transformation; MDS transformed into AML; acute monoblastic and monocytic leukemia (AMoL; FAB M5); and AML in relapse. The molecular pathogenesis and prognostic significance of isolated trisomy 2 remains unknown due to the small number of reported cases. Herein, we report 11 cases of isolated trisomy 2 in hematologic disorders seen in the Mayo Clinic Cytogenetics laboratory from 1996-2012. The majority of patients were older males (7/11) ranging in age from 63 to 93 years. The underlying bone marrow pathologic diagnoses include: hypercellular bone marrow without diagnostic features of malignancy (cases 1 and 2); MDS, refractory cytopenia with multilineage dysplasia (RCMD) subtype (cases 3 and 4); RAEB-1 (cases 5 and 6); long-standing history of primary myelofibrosis now with 7% bone marrow blasts (case 7); acute myeloid leukemia (AML), not otherwise specified (cases 8 and 9); AML with myelodysplasia-related changes (cases 10 and 11). Trisomy 2 has been suggested to represent an age-related phenomenon as it is seen predominantly in older individuals demonstrating this abnormality. Our data suggests that this could be a possible explanation since all of the eleven cases were ages 63 and over. Based on the limited clinical information in our study, it appears that isolated trisomy 2 harbors little prognostic significance and that, rather, the prognostic significance is driven by the underlying pathologic diagnosis. For example, 3 of the 4 AML cases and the case of PMF with increasing bone marrow blasts survived only 7, 8, 6 weeks and 21 weeks post bone marrow biopsy/cytogenetic evaluation, respectively. Although our study only has two cases that lack diagnostic features of malignancy, one of these cases survived 10 years following the identification of the cytogenetic abnormality. Therefore, trisomy 2 as a sole abnormality should not be considered as definitive evidence for MDS in the absence of diagnostic morphological criteria (similar to trisomy 8 and 20q deletion). Disclosures: No relevant conflicts of interest to declare.

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Telomerase Activity Is High In Essential Thrombocythemia and Polycythemia Vera, but Not In Myelofibrosis – a Comprehensive Analysis on Telomerase Activity and Cytogenetics In Myeloproliferative Neoplasm and Myelodysplastic Syndromes

Blood ◽

10.1182/blood.v116.21.4462.4462 ◽

2010 ◽

Vol 116 (21) ◽

pp. 4462-4462

Author(s):

Miyoung Kim ◽

Bora Oh ◽

Tae Young Kim ◽

Sang Mee Hwang ◽

Dong Soon Lee

Keyword(s):

Myelodysplastic Syndromes ◽

Cytogenetic Analysis ◽

Conflicts Of Interest ◽

Myeloproliferative Neoplasm ◽

Telomerase Activity ◽

Initial Diagnosis ◽

Conventional Cytogenetic Analysis ◽

Normal Controls ◽

Clonal Abnormalities

Abstract Abstract 4462 Introduction Telomere maintains genetic stability and cell replication by capping and protecting the end of the chromosomes. Telomere is synthesized by an enzyme called telomerase which contains a catalytic subunit, the human telomerase reverse transcriptase (hTERT). In this study, we compared the telomerase activity in two disease groups of distinctive pathophysiology - myeloproliferative neoplasm (MPN) and myelodysplastic syndromes (MDS) characterized by abnormally increased effective hematopoiesis and ineffective hematopoiesis resulting in intramedullary apoptosis, respectively. The result was also analyzed in association with the number of metaphase cells in conventional cytogenetic analysis. Materials and Methods Real time PCR to measure telomerase activity was performed using Quantitative Telomerase Detection kit (Allied Biotech, Inc., Ijamsville, MD) as per manufacturer¡&hibar;s instruction. The study included bone marrow cells of 99 normal controls, 114 with MPN (72 at initial diagnosis and 42 at follow up) and 73 with MDS patients (50 at initial diagnosis and 23 at follow up). MPN patients at initial diagnosis consisted of 22 CML in chronic phase, 18 ET, 11 PV, 12 PMF and 8 MPN, unclassifiable. MDS patients at initial diagnosis consisted of 19 RA or RCMD, 10 RAEB-1 and 13 RAEB-2. Results A trend of decreasing telomerase activity was observed as age increases in normal controls: 8239.19 (molecules/reaction) in age under 70yrs vs. 6669.46 in age over 70yrs (p>.05). Telomerase activity was higher in those with clonal chromosomal abnormalities detected in conventional cytogenetic analysis than those without clonal abnormalities (47600.82 vs. 11729.21, p>.05). In those with clonal abnormalities, patients with clonal abnormalities in >50.0% of metaphase cells analyzed showed significantly higher telomerase activity than patients with clonal abnormalities in <50.0% of metaphase cells (19183.06 vs. 6077.40, p=.014). Telomerase activity in MPN was significantly higher than that in normal controls (12990.12 vs. 8033.06, p=.027). A decreasing order of telomerase activity was observed in ET, PV and CML (20112.08, 17196.70 and 9835.25), and PMF showed significantly low telomerase activity than other subtypes (p=.005). In contrast, MDS showed slightly lower telomerase activity than normal controls, however, the difference was not statistically significant (7170.66 vs. 8033.06, p>.05). Telomerase activity was the highest in RAEB-2 and the lowest in RAEB-1 in between, without statistical significance (10532.24 and 6963.78, p=.441). Conclusion High telomerase activity in patients with high clonal cell proportion in conventional cytogenetic analysis could be explained by the hypothesis that cells with high proliferative activity form metaphase cells easily. High telomerase activity in MPN and low telomerase activity in MDS reflect the abnormally increased hematopoiesis and the ineffective hematopoiesis in each disease. ET and PV showed higher telomerase activity than CML, which could be explained by further analysis in line with clinical findings of the patients other than disease subtype. Low telomerase activity in PMF might result from the scarcity of normal hematopoietic cells and the high apoptotic fraction of megakaryocytes compared with anti-apoptotic profile of ET cells. Our result suggests that the potential of telomerase inhibitors should be investigated in patients with high telomerase activity. Disclosures: No relevant conflicts of interest to declare.

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U2AF1 mutations In Primary Myelofibrosis Cluster With Normal Karyotype and JAK2V617F and Are Strongly Associated With Anemia and Thrombocytopenia

Blood ◽

10.1182/blood.v122.21.4060.4060 ◽

2013 ◽

Vol 122 (21) ◽

pp. 4060-4060

Author(s):

Ayalew Tefferi ◽

Christy Finke ◽

Terra L Lasho ◽

Emnet A Wassie ◽

Ryan A Knudson ◽

...

Keyword(s):

Conflicts Of Interest ◽

Mutation Screening ◽

Prognostic Significance ◽

Multivariable Analysis ◽

Normal Karyotype ◽

Primary Myelofibrosis ◽

Prognostic Models ◽

International Prognostic Scoring System ◽

Pathway Gene ◽

Idh Mutations

Abstract Background Spliceosome pathway gene mutations are recurrent in myeloid malignancies with the highest frequencies reported for myelodysplastic syndromes associated with ring sideroblasts (MDS-RS; 85%), MDS without RS (44%) and chronic myelomonocytic leukemia (CMML; 55%) (Nature 2011;478:64). SF3B1 mutations were the most frequent (75%) in MDS-RS and SRSF2 mutations in CMML (28%); U2AF1 mutational frequencies were 12% in MDS without RS and 8% in CMML. We have previously described SRSF2 (Blood. 2012;120:4168) and SF3B1 (Leukemia. 2012;26:1135) mutations in 17% and 6.5% of patients with primary myelofibrosis (PMF); prognostic relevance was shown for the former but not the latter. Objectives The objectives of the current study were to i) describe the incidence of U2AF1 mutations in PMF and their correlation with clinical features, karyotype and other mutations and ii) examine the prognostic significance of U2AF1 mutations in PMF, in the context of both conventional prognostic models and other prognostically-relevant mutations. Methods Information on clinical and laboratory parameters and karyotype was available in all study patients, at time of referral, which coincided with time of sample collection for mutation screening. Risk stratification was according to the Dynamic International Prognostic Scoring System (DIPSS)-plus system. U2AF1 and other mutations were analyzed using standard PCR techniques and bidirectional sequencing; for U2AF1, two hot spots that included residues S34 and Q157 were amplified. Results A total of 251 PMF patients (median age 63 years; 160 males) were studied. DIPSS-plus risk distribution was high in 32%, intermediate-2 in 38%, intermediate-1 in 17% and low in 13% of the patients. The frequency of each DIPSS-plus adverse feature was as follows: age >65 years (42%), transfusion need (32%), hemoglobin <10 g/dL (47%), leukocyte count >25 x 10(9)/L, (16%), platelet count <100 x 10(9)/L (22%), ≥1% blasts (57%), constitutional symptoms (35%), and unfavorable karyotype (9%). Karyotype was normal in 156 (63%) patients. At a median follow-up of 48 months, 158 (63%) deaths and 27 (11%) leukemic transformations were recorded. Mutational frequencies Forty-one (16.3%) patients harbored U2AF1 mutations: 16 (39%) Q157P, 10 (24%) Q157R, 8 (20%) S34F, 4 (10%) S34Y and one each for Q157P/E159A, Q157R/S34Y and Q157-Y158insYE. Frequencies for other mutations were 11% for SRSF2, 7.3% for SF3B1, 31% for ASXL1, 5.5% for EZH2, 5% for IDH1/2 and 58% for JAK2V617F. U2AF1 mutations were usually, but not always, exclusive of other spliceosomal mutations: one patient expressed all three spliceosomal mutations. The frequency of any one of the three spliceosomal mutations was 34%. Clinical correlates U2AF1 mutations were significantly associated with older age (p=0.02), JAK2V617F (p=0.002), mutant ASXL1 (p=0.04), transfusion need (p<0.0001), hemoglobin <10 g/dL (p<0.0001), platelets <100 x 10(9)/L (p<0.0001) and normal karyotype (p=0.006). The associations with anemia, thrombocytopenia, JAK2V617F and normal karyotype were inter-independent; the frequency of U2AF1 mutations in the presence of anemia was 29%, transfusion need 36%, thrombocytopenia 35%, JAK2V617F 23% and normal karyotype 21%. Prognostic interactions U2AF1 mutations were associated with inferior overall (p=0.004) but not leukemia-free (p=0.6) survival. However, the survival association was fully accounted for by the above-mentioned clustering of U2AF1 mutations with anemia and thrombocytopenia. Similarly, although multivariable analysis of U2AF1, SRSF2, ASXL1, EZH2 and IDH mutations identified the first three as being independently predictive of poor survival, only ASXL1 and SRSF2 remained significant when either anemia or thrombocytopenia was included as a co-variate. Conclusions U2AF1 mutations are the most frequent spliceosome pathway mutations in PMF, cluster with normal karyotype and JAK2V617F, and are strongly associated with anemia and thrombocytopenia; the latter suggests a pathogenetic contribution to ineffective hematopoiesis in PMF. The current study also suggests that more than one third of patients with PMF carry a spliceosome mutation. Disclosures: No relevant conflicts of interest to declare.

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Cytogenetics of 125 Untreated Patients with Binet Stages B or C B-CLL. Preliminary Results of a Multicentric Study from the French Cooperative Study Group on CLL.

Blood ◽

10.1182/blood.v106.11.5003.5003 ◽

2005 ◽

Vol 106 (11) ◽

pp. 5003-5003

Author(s):

Florence Nguyen-Khac ◽

Nathalie Leporrier ◽

Isabelle Radford-Weiss ◽

Christine Lefebvre ◽

Christian Bastard ◽

...

Keyword(s):

Prospective Trial ◽

Prognostic Significance ◽

Cooperative Study Group ◽

Multicentric Study ◽

Variable Regions ◽

Mutational Status ◽

Morphological Criteria ◽

Trisomy 12 ◽

Igvh Mutational Status ◽

Balanced Translocations

Abstract Numerous cytogenetic and Fluorescence In Situ Hybridization (F) data have been previously published with the aim of defining prognosis subgroups of B-CLL patients. These studies included heterogeneous series of patients (treated or not, and at various stages of the disease). We have studied with conventional cytogenetic and F analyses an homogeneous cohort of 125 untreated Binet stages B or C B-CLL patients enrolled in a prospective trial on the basis of biological and morphological criteria, with a Matutes score 4/5. Karyotype (K) was systematically performed and F analysis carried out using four probes CEP12, 13q14, 11q22 (ATM), 17p13 (TP53) on metaphases and interphase nuclei. Out of 104 successful K, 65 (63%) showed abnormalities. Among them, 32% exhibited complex K, 25% balanced translocations and 35% unbalanced translocations. The most frequent abnormalities were 11q– (13%), +12 (13%), 13q– (10%), 6q– (8%), 17p– (5%), 14q32 rearrangement (5%), X or Y loss (4%), +19 (4%). Two translocations not previously reported involving the 14q32 locus were observed. F analyses were performed on 116 patients, 108 with the four probes, 8 with only 1–3 probes. 82 out of the 108 patients (76%) analyzed with 4 probes showed one or more abnormalities. Deletions of 13q were observed in 52%, ATM in 26%, TP53 in 10%, and trisomy 12 was present in 13%. Abnormal F patterns were observed in 51% of patients with normal K with at least one of the 4 probes, versus 92% of patients with abnormal K. Among abnormal K, F analyses detected additional cryptic deletions in 6/19 cases for ATM (31%), 29/39 for 13q14 (74%), and 4/9 for TP53 (45%). TP53 deletion was never detected among normal K, but in 15% of abnormal K. Among them, there were more TP53 deletion in complex K (24% vs 2%, p=0.005). Additional karyotypic abnormalities were found with a single F anomaly in 6/7 (86%) cases with del ATM, 12/17 (71%) with del13q, 4/6 (67%) with +12, 1/2 with del TP53. In the group of patients with normal F pattern, 81% had a normal K whereas 19% had an abnormal one (p<0.0001). A preliminary correlation with the immunoglobulin heavy chain variable regions (IgVH) mutational status was performed for 54 patients. The unmutated status (41/54, 76%) was more frequently associated with abnormal K (deletions 11q, ATM, and TP53, unbalanced translocations, and complex K), than mutated status. There were more del13q alone (25% vs 3%, p=0.01) and trisomy 12 in mutated patients when compared with unmutated ones. This preliminary study underscores the interest of performing both conventional cytogenetic (classically stimulated) and systematic F analysis in advanced stage CLL. Results are complementary and, as previously reported, display a better prognostic significance than F alone. Furthermore, the systematic use of the 6q21, 14q32 and CEP19 probes in F analyses should be recommended when the K is normal. Comparison with other relevant biological parameters (ZAP70, sCD23, CD38) is ongoing.

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Towards a Better Understanding of Epidemiology, Survival and Treatment in Myeloproliferative Neoplasms: Results of the European Leukemianet Registry (ERNEST study)

Blood ◽

10.1182/blood.v124.21.1849.1849 ◽

2014 ◽

Vol 124 (21) ◽

pp. 1849-1849 ◽

Cited By ~ 1

Author(s):

Tiziano Barbui ◽

Arianna Masciulli ◽

Marco Scarano ◽

Gianni Tognoni ◽

Sara Sisti ◽

...

Keyword(s):

Conflicts Of Interest ◽

Myeloproliferative Neoplasms ◽

Prognostic Significance ◽

Multivariable Analysis ◽

Cox Proportional Hazards ◽

Therapeutic Interventions ◽

Rank Test ◽

International Prognostic Scoring System ◽

Continuous Variables

Abstract Background The promising development of perspectives in innovative therapeutic interventions in myelofibrosis (MF) makes the establishment of large collaborative networks ready to assure reliable comparability of cases and increasingly relevant data. This will provide assurance regarding efficiency and consistency in the development of clinical and epidemiological knowledge. We report here the results of the pilot phase of ERNEST, whose acronym defines its purpose: European Registry for Myeloproliferative Neoplasms: towards a better understanding of Epidemiology, Survival and Treatment. This project promoted by the European LeukemiaNet (ELN) collaboration is coordinated by the Fondazione Mario Negri Sud (Italy) and supported by an unrestricted educational grant by Novartis. Patients and Methods In order to test the feasibility of a prospective epidemiological outcome-oriented registry across centers, expected to vary in the characteristics of their populations and care practices, a retrospective analysis was implemented based on a strictly pre-defined protocol. Patients with Primary Myelofibrosis (PMF), Post- Essential Thrombocythemia Myelofibrosis (PET-MF) and Post- Polycythemia Vera Myelofibrosis (PPV-MF) which were diagnosed in the participant centers between January 2001 and December 2012, with available follow-up information, were eligible for inclusion. Chi-square test and Mann-Whitney test were used to compare data at presentation by diagnosis for categorical and continuous variables respectively. Standard time-to-event methods were used for data analysis, including log-rank test, Kaplan-Meier survival graphs, and Cox proportional hazards models to calculate hazard ratios along with 95% confidence intervals (CI). Multivariable analysis was performed adjusting for unbalanced and relevant prognostic covariates. Results From February 2013 to May 2014, we received data of 1209 evaluable patients from 13 centers in 5 European countries (Italy, Germany, Spain, United Kingdom, Sweden): 61% were PMF, 20% PET-MF and 19% PPV-MF (median age: 66 years). 23%, 37% and 40% of diagnosis were performed between 2001-2004, 2005-2008 and 2009-2012 respectively. Variability was found for the presence of constitutional symptoms (from 43% in PET-MF to 49% in PPV-MF); an excess of splenomegaly emerged in PPV-MF cohort (84% vs 74% and 75% in PMF and PET-MF respectively). Mean value + SD of Hb, WBC and PLT were: 13.9 + 16.5 g/dl, 17.4 + 30.8 109/l, 372 + 316 109/l respectively with higher levels of Hb and WBC in PPV-MF patients than PET-MF and PMF (p<0.001). No variability was seen for presence of peripheral blasts and cytogenetic abnormalities. During follow-up (median duration: 2 years) 405 patients (33.5%) died without any differences among diagnosis subtypes. Leukemic transformation was experienced by 8% of the whole cohort (9% in PMF, 7% in PET-MF and 8%in PPV-MF). A multivariable’s Cox analysis was performed on the whole cohort including sex, diagnosis and International Prognostic Scoring System (IPSS) as covariates of interest. Besides male sex [Hazard Ratio (HR) 1.49 (95% CI (1.21-1.83), p<0.001] the prognostic significance of IPSS was confirmed with an HR 2.19 [(95% CI (1.64-2.92), p<0.001] for IPSS 2, and HR 4.20 [(95% CI (3.20-5.53), p<0.001] for IPSS >3, as compared with the reference category of patients with IPSS 0-1. As shown in Figure 1, an exploratory analysis documented different patterns of predictivity when the analysis was stratified according to the diagnosis subtypes. The determinants of the prognostic value of IPSS in PPV-MF vs PET-MF (and PMF, data not shown) would certainly deserve fully adjusted analysis in prospective well defined cohorts of patients. Conclusions The intensive quality control needed to assure the reliability, representativeness and the comparability of the data across international centers with expertise in this field confirms both the interest, but also the challenge of a cooperative epidemiological effort capable of representing a knowledge producing shared resource in the area of MF and other rare disease. Based also on the methodological and operational challenge resulting from this pilot study, a prospective study has been activated starting on September 2013. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

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EVALUATION of the Intra-LABORATORY Reproducibility of Percentage of Blast Counting IN MYELODYSPLASIA

Blood ◽

10.1182/blood.v116.21.4981.4981 ◽

2010 ◽

Vol 116 (21) ◽

pp. 4981-4981

Author(s):

Soraya Wuilleme ◽

Marion Eveillard ◽

Sophie Barille ◽

Francoise Accard ◽

Laurence Lodé ◽

...

Keyword(s):

Bone Marrow ◽

Scoring System ◽

Who Classification ◽

Conflicts Of Interest ◽

Scoring Systems ◽

Poor Quality ◽

International Prognostic Scoring System ◽

Inter Observer Variability ◽

Morphological Criteria ◽

Blast Cells

Abstract Abstract 4981 Background. In the two International Prognostic Scoring Systems of Myelodysplastic Syndrom (MDS), the percentage of BLAST cells in the Bone Marrow is the most important parameter implicated in score: either directly in the IPSS (International Prognostic Scoring System) (1) or indirectly in the WPSS (Who classification-based prognostic scoring system) (2). In these two systems, as in the recommendations of the WHO 2001 (3), the morphological criteria defining blasts cells compared to promyelocyte cells are not specified. In 2005, the IWGM-MDS (International Working Group on Morphology of myelodysplastic syndrome) (4-6) has established morphological criteria defining Blasts cells. PURPOSE OF THE WORK. Our objective in this study is to evaluate the reproducibility, among five observers of our laboratory, of the counting of the marrow blasts of 73 myelodysplasia. Materials and Methods. This study was conducted in several stages. 1st step was to test the implementation of the MDS-Foundation (http://www.mds- foundation.org/virtualmicroscopy) by 5 observers. The 2nd step was to perform correlation test of the selected cells from RAEB (selected and stained in our laboratory conditions), for the 5 observers. Finally Step 3 was to assess the correlation of blast percentage of 73 bone marrow smears from MDS patients, selected due to the presence of an excess of blast cells in the first reading on the bone aspiration. Results. Our results show that the correlation on counting blasts is generally satisfactory (percentage agreement: Test 1 = 86% and test 2 = 94%), while the concordance on the counting blasts of bone marow smears of 73 MDS patients and concordance on the WHO classification seems less satisfactory (agreement 3/5 observers. = 95% but agreed to 4–5/5 observers = 64%). These results can be explained partly by the inter-observer variability and by the variability of some parameters specific to smear marrow (poor quality smears, the staining and/or poverty of smears). Conclusion. The evaluation of the blasts in the MDS must be achieved: (i) at least 500 cells counted (ii) by at least two different observers (iii) by a third observer in discordant case. Despite these recommendations, the assessment of the blasts in myelodysplasia is difficult to achieve in many cases. Disclosures: No relevant conflicts of interest to declare.

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Cytogenetic Analysis in Patients with Newly Diagnosed Myelodysplastic Syndromes in Southern Italy

Blood ◽

10.1182/blood.v120.21.5200.5200 ◽

2012 ◽

Vol 120 (21) ◽

pp. 5200-5200

Author(s):

Esther Natalie Oliva ◽

Francesco Albano ◽

Nicola Di Renzo ◽

Vincenzo Pavone ◽

Stefano Molica ◽

...

Keyword(s):

Bone Marrow ◽

Myelodysplastic Syndromes ◽

Cytogenetic Analysis ◽

Southern Italy ◽

Conflicts Of Interest ◽

Refractory Anemia ◽

Newly Diagnosed ◽

Conventional Cytogenetic Analysis ◽

Conventional Analysis ◽

Italian Regions

Abstract Abstract 5200 Background: Bone marrow karyotype in myelodysplastic syndromes (MDS) is essential to define the prognosis and to guide treatment decisions, including targeted therapies. Due to the lack of an extensive national MDS registry in Italy, epidemiological data on MDS, including cytogenetics, throughout the territory is unknown. Objective: We evaluated the incidence of cytogenetic abnormalities amongst newly diagnosed MDS patients in in 2 southern Italian regions (Calabria and Puglia). Methods: A pilot project, denominated ANDROMEDA (ANalysis of cytOgenetics alteRatiOn in the MyEloDysplAstic syndromes) was developed in 17 centers to offer a service of conventional cytogenetic analysis for all consecutive patients undergoing diagnostic evaluation for cytopenia and suspected MDS between January 1 and December 31, 2011. The study conformed to the ethical standards set out in the Declaration of Helsinki and was approved by institutional review boards at each participating center. Patients were required to provide their written informed consent. Clinical characteristics of patients and bone marrow morphology, iron staining and histology were registered. Bone marrow samples were centralized for standard cytogenetic studies and fluorescence in situ hybridization to two dedicated genetics laboratories (one for each region), blind to patients' data. Results: Two hundred and thirty-five patients were evaluated and MDS diagnosis was confirmed in 220 cases (88. 3%), according to WHO criteria. The overall incidence of clonal chromosome abnormalities detected by conventional analysis was 36. 9%. Single abnormalities included +8 (13 cases, 5. 8%), del(5q) (12 cases, 5. 4%), –Y (11 cases, 5. 0%) and del(7)/-7 (4 cases, 1. 8%). Complex karyotypes were detected in 18 (8. 1%) cases. Among all cases only 10 (4. 5%) bone marrow samples were not evaluable for cytogenetic analysis. FISH revealed additional abnormalities not identified by conventional analysis only in 3 (1. 3%) out of 72 cases. Patients were classified in WHO subtypes: 39. 2% refractory cytopenia with unilineage dysplasia (RCUD), 1. 5% refractory anemia with ring sideroblasts (RARS), 32. 5% refractory cytopenia with multilineage (RCMD), 10. 8% refractory anemia with excess of blast-1 (RAEB-1), 9. 3% refractory anemia with excess of blast-2 (RAEB-2), 4. 1% MDS with deletion 5q (MDS 5q-) and 2. 6% MDS unclassifiable (MDS-U). Conclusions: These preliminary results demonstrate that the incidence of abnormal karyotype patterns and WHO subgroups in MDS patients in Southern Italy is comparable with that described in other geographical areas. It is confirmed that conventional cytogenetic analysis is a standard in the diagnostic workup of MDS of patients with a suspected myeloid malignancy in order to identify primary abnormalities and prognostic models. Disclosures: No relevant conflicts of interest to declare.

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Prognostic Implications of Cytogenetic Features in Myelodysplastic Syndromes

Oncology & Hematology Review (US) ◽

10.17925/ohr.2013.09.1.60 ◽

2013 ◽

Vol 09 (01) ◽

pp. 60 ◽

Cited By ~ 1

Author(s):

Carolina B Belli ◽

Yesica Bestach ◽

Laura Kornblihtt ◽

Irene B Larripa ◽

◽

...

Keyword(s):

Clinical Practice ◽

Myelodysplastic Syndromes ◽

Cytogenetic Analysis ◽

Prognostic Significance ◽

Cytogenetic Response ◽

International Prognostic Scoring System ◽

Risk Indicator ◽

Intermediate Group ◽

The Past ◽

Prognostic Implications

Myelodysplastic Syndromes (MDS) are a heterogeneous group of hematologic diseases characterized by refractory cytopenia(s) and variable risk of leukemic progression. Cytogenetic analysis is important in day-to-day clinical practice helping to define subgroups of MDS patients who share similarities in the course of the disease. There are recurring aberrations affecting chromosomes 5, 7, 8, and 20. While all of them do not suggest a therapeutic approach, their presence has been considered as a risk indicator since the original international prognostic scoring system (IPSS) was published. The most recent cytogenetic stratifications tried to find the prognostic significance of less frequent alterations which have been longer included in the intermediate group. Moreover, monitoring of karyotype changes is suggested to evaluate cytogenetic response to treatments and the acquisition of new aberrations associated to an unfavorable outcome. This review focuses on different cytogenetic risk stratifications that have been published during the past 20 years and the molecular background of the most relevant chromosomal findings.

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