- Email: [email protected]
P024 Computerized nuclear texture analysis for the characterization of atypic immature cells in MDS
2. Diagnosis and Prognostic Markers P022 Serum CD44 levels predict survival in patients with low-risk myelodysplastic syndromes R. Stauder1 ° , U. Germing2 , W. Sperr3 , P. Valent3 , H. Zwierzina4 , H. Ulmer5 , J. Loeffler-Ragg4 . 1 Department of Internal Medicine V, Medical University, Innsbruck, Innsbruck, Austria; 2 Heinrich-Heine-University, Germany; 3 Medical University of Vienna, Austria; 4 Medical University, Innsbruck, Austria; 5 Innsbruck Medical University, Austria *E-mail: [email protected] Aberrant expression of the adhesion molecule CD44 correlates with poor prognosis in various neoplasms. To evaluate the prognostic impact of CD44 in myelodysplastic syndromes (MDS) serum levels of soluble CD44 (sCD44) were measured in 130 MDS patients using an enzymelinked immunosorbent assay (ELISA). sCD44 levels were significantly elevated in MDS patients compared to healthy donors (p < 0.05), and were found to correlate with distinct FAB subtypes. The highest levels of sCD44 were found in patients with CMML and in patients with MDS transformed into acute myeloid leukemia (AML). In univariate analysis elevated levels of sCD44 s were significantly correlated with shorter overall survival in MDS-patients (9 versus 37 months; p < 000). In multivariate analysis sCD44 s displayed prognostic significance for survival independent from the International Prognosis Scoring System (IPSS). To test for refined prognostication, each IPSS risk group was split into two separate categories based on sCD44 s levels. Using this approach, MDS patients with a shorter survival were identified both in the IPSS low-risk (p = 0.014) and in the IPSS Int-1 group (p = 0.031). The CD44 s-adjusted IPSS defines a cohort of MDS patients with unfavorable prognosis within the low and intermediate-1 IPSS groups, which might be helpful in risk stratification and in therapeutic algorithms.
P023 Lymphoid micromegakarycytes is a unfavourable prognostic factor in patients with primary myelodysplastic syndromes Z. Xiao ° , W. Cui, L. Li, Y. Zhang, T. Qin. The State Key Laboratory of Experimental Hematology, Department of Clinical Hematology, Institute of Hematology and Blood Diseases Hospital, CAMS, Tianjin, China *E-mail: [email protected] Purpose: Several studies have demonstrated that the most valid prognostic scoring system used in MDS is the IPSS. Recently, a new prognostic scoring system, WPSS, was proposed based on the WHO classification. The prognostic significance of WPSS in Chinese patients with MDS was evaluated and a modified WPSS was provided.
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Methods: In order to assess the role of the main prognostic factors in MDS, 164 adult patients were retrospectively analyzed. Results: The median follow-up time was 19(1–138) months and the median survival was 36 months. 2-year survival rate was 60% and 5-year survival rate was 42%. In patients with very low-risk, low-risk, intermediate-risk, high-risk and very high-risk stratified by WPSS, 2-year survival was 100%, 96%, 81%, 38% and 14%, respectively; 5-year survival was 100%, 83%, 54%, 20% and 0, respectively (P < 0.001). Among the parameters, elevated MCV, MCH, N-ALP score and PAS negative were good prognostic factors while decreased Hb, BPC and blast more than 5%, dysplasia more than 1 lineage and lymphoid micromegakaryocyte (MEGly) presented in bone marrow, elevated LDH in serum were unfavourable prognostic factors by uni-variable analysis, and MCV, MEGly and Blast more than 5% had independent prognostic significance by multi-variable analysis (P = 0.011, 0.013 and 0.016, respectively). WPSS was modified by removing chromosomal karyotype and transfusion dependence and adding in MCV and MEGly, and the patients were stratified into low-risk, intermediaterisk and high-risk groups according to the scores. Patients with low-risk, intermediate-risk and high-risk stratified by modified WPSS, 2-year survival was 94%, 68% and 49%, respectively; 5-year survival was 86%, 53% and 14%, respectively(P < 0.001). Conclusions: WPSS based on WHO classification was adapted to prognostic determination in Chinese patients with MDS. Lymphoid micromegakaryocyte had independent prognostic significance in MDS patients.
P024 Computerized nuclear texture analysis for the characterization of atypic immature cells in MDS I. Lorand-Metze ° , J. Vido, R. Adam, K. Metze. State University of Campinas, Brazil *E-mail: [email protected] Purpose: Bone marrow (BM) cytology is essential for the diagnosis of MDS. However, in cases with many atypical cells it may be difficult to quantify precisely myeloblasts and promyelocytes. Our aim was to investigate whether computerized image analysis of routine cytology would be able to characterize these cell types. Methods: In MGG stained BM smears of 14 newly diagnosed MDS (8 RCMD, 4 RAEB, 2 RA) patients and 15 cases of normal BM, blasts, promyelocytes and atypic precursors were digitalized and interactively segmented. The classification of the cells was done by consensus of two observers. Nuclear morphometry and texture features derived from the co-occurrence matrix (Haralick features), fractal dimensions and Fast-Fourier-transformed (FFT) images were calculated.
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Results: Form factors, gray levels, and 10 texture features were different between the cell types. Focusing on the atypical immature cells, the values of 10 variables were close to that of promyelocytes, one feature was very similar to that of blasts, and finally 8 texture features showed intermediate values between blasts and promyelocytes. Conclusion: Bone marrow atypical immature cells are sometimes very difficult to classify correctly in routine cytology. Our study shows that these cells present nuclear texture features intermediate between blasts and promyelocytes. They could be abnormally maturating blasts or atypical promyelocytes. This issue is critical for the correct classification and prognosis of MDS. Computerized image analysis could be helpful in order to overcome this problem. In cases with a high degree of cell atypias it may be difficult to distinguish myeloblasts from promyelocytes. Consensus criteria have been established, but in daily practice there may remain difficulties in classifying individual cells. Supported by CNPq and FAPESP.
P025 Gain of 1q as a potential adverse prognostic marker in myelodysplastic syndrome D.S Lee1 ° , H.-K. Park1 , H.R. Lee1 , H.I. Cho1 , H.K. Kim1 , S.S. Yoon2 , B.K. Kim2 . 1 Department of Laboratory Medicine Seoul National University College of Medicine, Jongno-gu, Korea; 2 Department of Internal Medicine Seoul National University College of Medicine, Korea *E-mail: [email protected] Background: The gain of the 1q region, which is a recurrent chromosomal aberration in B lymphoproliferative disorder, has been reported one of the most common anomalies in Korean myelodysplastic patients. Recently, risk based application of hypomethylating agents or tailored therapy in MDS rely on the prognostic variables of International Prognostic Scoring System (IPSS). To investigate the possibility of 1q gain as a new prognostic marker, we evaluated the prognostic impact of 1q gain, along with comparison with IPSS variables. Methods: A total of 117 patients with newly diagnosed MDS between 1997 and 2007 at the Seoul National University Hospital were investigated. Fluorescence in situ hybridization (FISH) studies with 5 specific probe (EGR1 for 5q31 deletion, D7S522 for 7q31 deletion, CEP8, D20S108 for 20q12 deletion, LSI 1p36/1q25 for 1q gain) and conventional G-banding karyotyping were performed on bone marrow aspirates. Other laboratory findings, such as hemoglobin (Hb), absolute neutrophil count (ANC), platelet count, bone marrow blast percent and IPSS score, and clinical data were collected through the individual medical records. Results: The median age was 54 years and the maleto-female ratio was 1.4. Using WHO classification,
refractory anemia(RA) was 27.4% and the other subgroups as follows: RA with ringed sideroblasts(RARS), 3.4%; refractory cytopenia with multilineage dysplasia(RCMD), 8.5%; RCMD with ringed sideroblasts(RCMD-RS), 0.9%; RA with excess blasts-1(RAEB-1), 26.5%; RAEB-2, 31.6%; and 5q− syndrome, 1.7%. Cytogenetic abnormalities by FISH and G-banding were detected in 58 patients (49.6%). Most frequent anomaly was trisomy 8 occurring in 28 patients (23.9% of the 117 patients, 48.3% of the 58 patients with clonal cytogenetic abnormalities). Gain of 1q was the second common anomalies seen in 18 patients (15.4%) and other anomalies were −7/del7q (13.7%), −5/del5q (13.7%), and del20q (2.6%). G-banding showed gain of 1q in 7 cases, additional 11 patients with gain of 1q were revealed by FISH only. Patients with 1q gain showed a poor survival (median survival 23 months; n = 18) compared to patients without 1q gain (median survival 60 months; p = 0.02). EGR1 and D7S522 deletion by FISH also had a shorter median survival (8 months vs. 60 months p = 0.0001, 16 months vs. 60 months p = 0.005). The initial platelet count and blast count were found to affect overall survival, whereas CEP8 FISH, D20S108 FISH, Hb and ANC did not. Discussion: Our results show that gain of 1q is associated with an adverse clinical outcome and can be considered as a poor cytogenetic risk factor of IPSS. In the Western study, the prevalence of 1q gain was low because most studies report G-banding result only. But it may be increased up to 2.5 fold higher by using FISH analysis in combination with G-banding. A gain of 1q could be a candidate as an adverse prognostic marker in clinical practice, which could help for risk-adapted therapies.
P026 A limited benefit of isolated 5q deletion on prognosis − results of a long term retrospective analysis J. Cermak ° , D. Mikulenkova, J. Brezinova, K. Michalova. Institute of Hematology and Blood Transfusion, Praha, Czech Republic *E-mail: [email protected] A retrospective analysis of a long term outcome was performed in a cohort of 30 patients with isolated 5q deletion treated in Institute of Hematology in Prague before lenalidomide era in the years 1983–2006. Eleven patients fulfilled WHO diagnostic criteria of 5q− syndrome (5q−sy), 19 patients had isolated deletion of 5q (isoldel5q) not connected with typical 5q− syndrome. Median survival of patients with 5q−sy of 57.0 months was significantly different from that of patients with isoldel5q (18 months, p = 0.02). Estimated 3 year, 5 year and 10 year overall survival were 82%, 73% and 64% for 5q−sy and 37%, 26% and 21% for isoldel5q. A disease progression towards advanced MDS with excess of blasts was observed in 11 out of 19 patients (58%) with isoldel5q in contrast
P023 Lymphoid micromegakarycytes is a unfavourable prognostic factor in patients with primary myelodysplastic syndromes Z. Xiao ° , W. Cui, L. Li, Y. Zhang, T. Qin. The State Key Laboratory of Experimental Hematology, Department of Clinical Hematology, Institute of Hematology and Blood Diseases Hospital, CAMS, Tianjin, China *E-mail: [email protected] Purpose: Several studies have demonstrated that the most valid prognostic scoring system used in MDS is the IPSS. Recently, a new prognostic scoring system, WPSS, was proposed based on the WHO classification. The prognostic significance of WPSS in Chinese patients with MDS was evaluated and a modified WPSS was provided.
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Methods: In order to assess the role of the main prognostic factors in MDS, 164 adult patients were retrospectively analyzed. Results: The median follow-up time was 19(1–138) months and the median survival was 36 months. 2-year survival rate was 60% and 5-year survival rate was 42%. In patients with very low-risk, low-risk, intermediate-risk, high-risk and very high-risk stratified by WPSS, 2-year survival was 100%, 96%, 81%, 38% and 14%, respectively; 5-year survival was 100%, 83%, 54%, 20% and 0, respectively (P < 0.001). Among the parameters, elevated MCV, MCH, N-ALP score and PAS negative were good prognostic factors while decreased Hb, BPC and blast more than 5%, dysplasia more than 1 lineage and lymphoid micromegakaryocyte (MEGly) presented in bone marrow, elevated LDH in serum were unfavourable prognostic factors by uni-variable analysis, and MCV, MEGly and Blast more than 5% had independent prognostic significance by multi-variable analysis (P = 0.011, 0.013 and 0.016, respectively). WPSS was modified by removing chromosomal karyotype and transfusion dependence and adding in MCV and MEGly, and the patients were stratified into low-risk, intermediaterisk and high-risk groups according to the scores. Patients with low-risk, intermediate-risk and high-risk stratified by modified WPSS, 2-year survival was 94%, 68% and 49%, respectively; 5-year survival was 86%, 53% and 14%, respectively(P < 0.001). Conclusions: WPSS based on WHO classification was adapted to prognostic determination in Chinese patients with MDS. Lymphoid micromegakaryocyte had independent prognostic significance in MDS patients.
P024 Computerized nuclear texture analysis for the characterization of atypic immature cells in MDS I. Lorand-Metze ° , J. Vido, R. Adam, K. Metze. State University of Campinas, Brazil *E-mail: [email protected] Purpose: Bone marrow (BM) cytology is essential for the diagnosis of MDS. However, in cases with many atypical cells it may be difficult to quantify precisely myeloblasts and promyelocytes. Our aim was to investigate whether computerized image analysis of routine cytology would be able to characterize these cell types. Methods: In MGG stained BM smears of 14 newly diagnosed MDS (8 RCMD, 4 RAEB, 2 RA) patients and 15 cases of normal BM, blasts, promyelocytes and atypic precursors were digitalized and interactively segmented. The classification of the cells was done by consensus of two observers. Nuclear morphometry and texture features derived from the co-occurrence matrix (Haralick features), fractal dimensions and Fast-Fourier-transformed (FFT) images were calculated.
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Posters
Results: Form factors, gray levels, and 10 texture features were different between the cell types. Focusing on the atypical immature cells, the values of 10 variables were close to that of promyelocytes, one feature was very similar to that of blasts, and finally 8 texture features showed intermediate values between blasts and promyelocytes. Conclusion: Bone marrow atypical immature cells are sometimes very difficult to classify correctly in routine cytology. Our study shows that these cells present nuclear texture features intermediate between blasts and promyelocytes. They could be abnormally maturating blasts or atypical promyelocytes. This issue is critical for the correct classification and prognosis of MDS. Computerized image analysis could be helpful in order to overcome this problem. In cases with a high degree of cell atypias it may be difficult to distinguish myeloblasts from promyelocytes. Consensus criteria have been established, but in daily practice there may remain difficulties in classifying individual cells. Supported by CNPq and FAPESP.
P025 Gain of 1q as a potential adverse prognostic marker in myelodysplastic syndrome D.S Lee1 ° , H.-K. Park1 , H.R. Lee1 , H.I. Cho1 , H.K. Kim1 , S.S. Yoon2 , B.K. Kim2 . 1 Department of Laboratory Medicine Seoul National University College of Medicine, Jongno-gu, Korea; 2 Department of Internal Medicine Seoul National University College of Medicine, Korea *E-mail: [email protected] Background: The gain of the 1q region, which is a recurrent chromosomal aberration in B lymphoproliferative disorder, has been reported one of the most common anomalies in Korean myelodysplastic patients. Recently, risk based application of hypomethylating agents or tailored therapy in MDS rely on the prognostic variables of International Prognostic Scoring System (IPSS). To investigate the possibility of 1q gain as a new prognostic marker, we evaluated the prognostic impact of 1q gain, along with comparison with IPSS variables. Methods: A total of 117 patients with newly diagnosed MDS between 1997 and 2007 at the Seoul National University Hospital were investigated. Fluorescence in situ hybridization (FISH) studies with 5 specific probe (EGR1 for 5q31 deletion, D7S522 for 7q31 deletion, CEP8, D20S108 for 20q12 deletion, LSI 1p36/1q25 for 1q gain) and conventional G-banding karyotyping were performed on bone marrow aspirates. Other laboratory findings, such as hemoglobin (Hb), absolute neutrophil count (ANC), platelet count, bone marrow blast percent and IPSS score, and clinical data were collected through the individual medical records. Results: The median age was 54 years and the maleto-female ratio was 1.4. Using WHO classification,
refractory anemia(RA) was 27.4% and the other subgroups as follows: RA with ringed sideroblasts(RARS), 3.4%; refractory cytopenia with multilineage dysplasia(RCMD), 8.5%; RCMD with ringed sideroblasts(RCMD-RS), 0.9%; RA with excess blasts-1(RAEB-1), 26.5%; RAEB-2, 31.6%; and 5q− syndrome, 1.7%. Cytogenetic abnormalities by FISH and G-banding were detected in 58 patients (49.6%). Most frequent anomaly was trisomy 8 occurring in 28 patients (23.9% of the 117 patients, 48.3% of the 58 patients with clonal cytogenetic abnormalities). Gain of 1q was the second common anomalies seen in 18 patients (15.4%) and other anomalies were −7/del7q (13.7%), −5/del5q (13.7%), and del20q (2.6%). G-banding showed gain of 1q in 7 cases, additional 11 patients with gain of 1q were revealed by FISH only. Patients with 1q gain showed a poor survival (median survival 23 months; n = 18) compared to patients without 1q gain (median survival 60 months; p = 0.02). EGR1 and D7S522 deletion by FISH also had a shorter median survival (8 months vs. 60 months p = 0.0001, 16 months vs. 60 months p = 0.005). The initial platelet count and blast count were found to affect overall survival, whereas CEP8 FISH, D20S108 FISH, Hb and ANC did not. Discussion: Our results show that gain of 1q is associated with an adverse clinical outcome and can be considered as a poor cytogenetic risk factor of IPSS. In the Western study, the prevalence of 1q gain was low because most studies report G-banding result only. But it may be increased up to 2.5 fold higher by using FISH analysis in combination with G-banding. A gain of 1q could be a candidate as an adverse prognostic marker in clinical practice, which could help for risk-adapted therapies.
P026 A limited benefit of isolated 5q deletion on prognosis − results of a long term retrospective analysis J. Cermak ° , D. Mikulenkova, J. Brezinova, K. Michalova. Institute of Hematology and Blood Transfusion, Praha, Czech Republic *E-mail: [email protected] A retrospective analysis of a long term outcome was performed in a cohort of 30 patients with isolated 5q deletion treated in Institute of Hematology in Prague before lenalidomide era in the years 1983–2006. Eleven patients fulfilled WHO diagnostic criteria of 5q− syndrome (5q−sy), 19 patients had isolated deletion of 5q (isoldel5q) not connected with typical 5q− syndrome. Median survival of patients with 5q−sy of 57.0 months was significantly different from that of patients with isoldel5q (18 months, p = 0.02). Estimated 3 year, 5 year and 10 year overall survival were 82%, 73% and 64% for 5q−sy and 37%, 26% and 21% for isoldel5q. A disease progression towards advanced MDS with excess of blasts was observed in 11 out of 19 patients (58%) with isoldel5q in contrast