- Email: [email protected]
Cell cycle studies in myelodysplastic syndrome
3. MDS." Diagnostic Criteria and Cell Cycle
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platelets 13,000/btl, WBC 4,800/btl (neutrophils 14%, metamyelocytes 2%, myelocytes 1%, blasts 6%, eosinophils 3%, monocytes 21%, lymphocytes 53% and nucleated red blood cells 1%). The bone marrow was hypercellular with reduced megakaryocytes, 18.75% blasts without Auer rods, 5.5% monocytes and 28.75% erythroid progenitors. Dysplastic changes were found in all three lineages. Bone marrow cytogenetic analysis detected 45,XY,-7 [11]/ 45,XY, t(3 ;21)(q26; q22),-7 [ 11 ]. Progenitor cultures of bone marrow and peripheral blood showed poor growth and no GMCSF hypersensitivity. Disease was concluded as RAEB-t. Neither relative nor suitable unrelated donor was found. Patient underwent splenectomy and was treated with 6-MP. Morphological analysis of the spleen was compatible with the diagnosis of CMML. Records of medical history revealed recurrent otitis and bronchitis during the infant period together with IgA deficiency. Hepatosplenomegaly was found at the age of 4 months, when his Hb was 118 g/dl, platelets 276,000/btl, WBC 34,400/btl with 21% monocytes but no blasts. Bone marrow was hypercellular without increased blasts. Cytogenetic analysis was not performed. The boy was followed up until the age of 8 years at a local hematological center without any treatment. Diagnosis NF-1 was established at the age of 6 years. At the age of 8 years his spleen was enlarged 3 cm below costal margin, Hb was 115 g/dl, platelets 240,000/btl, WBC 15,200/btl (neutrophils 47%, eosinophils 5%, monocytes 4%, lymphocytes 46%). After a 6 year' s gap of follow-up, the disease flared up as RAEB-t. Allogeneic stem cell transplantation was performed 7 months from diagnosis. Patient died due to infection 6 months later. We present an unusual case of a patient with NF-1 who suffered from JMML with spontaneous improvement in the early childhood and relapsed with MDS/MPS overlap disease with monosomy 7 resembling RAEB-t or CMML at the age of 14 years.
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Hospital, Nashville, TN, USA; 4Esoterix Center for Innovation, Brentwood, TN, USA *E-mail: [email protected] To seek better diagnostic tools in Myelodysplastic syndrome (MDS) and a better understanding of its pathogenesis, we analyzed cell cycle parameters, DNA damage, and apoptosis in individual marrow precursor compartments using multiparametric flow cytometry in a series of MDS patients (n= 14). Diagnoses were confirmed and results analyzed with blinded review. Controls were patients lacking primary marrow disease. Flow analytes included CD45, SS, CD34, CD71, DRAQ 5 (DNA content), MPM-2 phosphorylation (mitoses), H2A.X phosphorylation (yH2A.X, phosphorylated by ATM in the presence of double strand DNA breaks), and Annexin V. Use of DRAQ 5 allowed analysis of DNA content in individual progenitor cell subsets (erythroid, myeloid, lymphoid, and stem cells); cycle results varied unexpectedly in individual subsets. Use of MPM-2 allowed confirmation of successful progression through the cell cycle (mitotic division), and simultaneous assessment of other parameters allowed this analysis in individual cell subsets. MDS patients demonstrated a significant increase in the G2 fraction in myeloid progenitors, with trends in S phase in myeloids and to a lesser extent post-G1 fractions in erythroid progenitors, with no proportionate increase in mitoses, yH2A.X binding was increased in erythroid and myeloid progenitors in MDS, with more modest elevation of Annexin V reactivity, yH2A.X and Annexin V reactivity varied with cell cycle compartment and lineage, and together were suggestive of DNA damage. Results suggest that MDS is characterized by DNA damage in hematopoietic progenitors, with post-G1 arrest or slowing of cell cycle progression, resulting in hypercellular marrow but failed marrow production of peripheral effector cells. These methods are useful for study of MDS biology. The abnormalities described may be useful as diagnostic tools.
M I N I M A L C R I T E R I A FOR DIAGNOSIS OF MDS
J. Bennett 1 *, G. Mufti 2. ICancer Center, University of Rochester, Rochester, NY, USA; 2Department of Hematology, King' s College, London; The MDS Foundation, Crosswicks, NJ, USA *E-mail: j [email protected] The W.H.O. has defined the minimal criteria for Dysplasia of cell lines as >10% dysplasia. However, the precise descriptions were lacking. An international morphology committee met on three occasions in 2005 and reviewed many cases of MDS. Criteria were established both for types of blasts; ringed sideroblasts; hypogranularity of neutrophils; megakaryoctic dysplasia (small nuclei and multiple nucleoli and megaloblastic erythroid forms. Each observer recorded the data electronically. In addition over 200 marrow cells were classified by each participant from the same images on a web site. The data is being analysed by a biostatistician for concordance and reproducibility and will be presented at the meeting. In addition plans are underway to organize a registry for Idiopathic Cytopenias of Unknown Significance (ICUS) for those patients who do not meet the minimal criteria.
[•-] CELL CYCLE STUDIES
IN M Y E L O D Y S P L A S T I C
SYN D ROME D.R. Head 1 *, J.W. Jacobberger 2, M. Jagasia 1, S. Goodman 1'3, L. Flye4, S. McClintock-Treep 1, E Boulos 1, E Jiang 1, G. Stelzer4, R. Briggs 1, K. Shults 4. 1Vanderbilt University Medical Center,
Nashville, TN, USA; 2Case Comprehensive Cancer Center, Cleveland, OH, USA; 3Nashville Veterans Administration
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M Y E L O D Y S P L A S T I C S Y N D R O M E S IN C H I L D H O O D : EPIDEMIOLOGICAL ASPECTS
M. Hamaji 1 *, L.F. Lopes 1, M. do Rosario Latorre 2. 1Department
of Pediatrics, Hospital A. C. Camargo, S6o Paulo, Brazi# 2Department of Epidemiology, Universidade de S6o Paulo, S6o Paulo, Brazil *E-mail: [email protected] The data most commonly found on the etiology of the myelodysplastic syndromes (MDS) in children are based on genetic alterations, while there are few studies on exposure to toxic substances of patients and their parents. The possible etiology of environmental factors in MDS in childhood is unknown, and the influence of exposure to pesticides in adolescent and children has been argued. The main objective of this work was to take the personal and familial history of pediatric patients with MDS and compare them with data of children with acute lymphocytic leukemia (ALL), children with acute myeloid leukemia (AML) and children with no neoplasia. A questionnaire was elaborated and applied through direct interview and, in rare cases, by telephone. 150 children were interviewed: 50 with no neoplasia, 40 MDS, 50 ALL and 10 AML. There was a statistically significant association between skin color and diagnosis (p =0.031, with 100% of children with AML being Caucasian against 67% of those with MDS), birthweight (p = 0.039, with highest prevalence of low birthweight in MDS), previously having lived in an agricultural zone (p=0.003, with highest prevalence in those with AML, 20%, against 0% in those with
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platelets 13,000/btl, WBC 4,800/btl (neutrophils 14%, metamyelocytes 2%, myelocytes 1%, blasts 6%, eosinophils 3%, monocytes 21%, lymphocytes 53% and nucleated red blood cells 1%). The bone marrow was hypercellular with reduced megakaryocytes, 18.75% blasts without Auer rods, 5.5% monocytes and 28.75% erythroid progenitors. Dysplastic changes were found in all three lineages. Bone marrow cytogenetic analysis detected 45,XY,-7 [11]/ 45,XY, t(3 ;21)(q26; q22),-7 [ 11 ]. Progenitor cultures of bone marrow and peripheral blood showed poor growth and no GMCSF hypersensitivity. Disease was concluded as RAEB-t. Neither relative nor suitable unrelated donor was found. Patient underwent splenectomy and was treated with 6-MP. Morphological analysis of the spleen was compatible with the diagnosis of CMML. Records of medical history revealed recurrent otitis and bronchitis during the infant period together with IgA deficiency. Hepatosplenomegaly was found at the age of 4 months, when his Hb was 118 g/dl, platelets 276,000/btl, WBC 34,400/btl with 21% monocytes but no blasts. Bone marrow was hypercellular without increased blasts. Cytogenetic analysis was not performed. The boy was followed up until the age of 8 years at a local hematological center without any treatment. Diagnosis NF-1 was established at the age of 6 years. At the age of 8 years his spleen was enlarged 3 cm below costal margin, Hb was 115 g/dl, platelets 240,000/btl, WBC 15,200/btl (neutrophils 47%, eosinophils 5%, monocytes 4%, lymphocytes 46%). After a 6 year' s gap of follow-up, the disease flared up as RAEB-t. Allogeneic stem cell transplantation was performed 7 months from diagnosis. Patient died due to infection 6 months later. We present an unusual case of a patient with NF-1 who suffered from JMML with spontaneous improvement in the early childhood and relapsed with MDS/MPS overlap disease with monosomy 7 resembling RAEB-t or CMML at the age of 14 years.
[
•
-
]
Hospital, Nashville, TN, USA; 4Esoterix Center for Innovation, Brentwood, TN, USA *E-mail: [email protected] To seek better diagnostic tools in Myelodysplastic syndrome (MDS) and a better understanding of its pathogenesis, we analyzed cell cycle parameters, DNA damage, and apoptosis in individual marrow precursor compartments using multiparametric flow cytometry in a series of MDS patients (n= 14). Diagnoses were confirmed and results analyzed with blinded review. Controls were patients lacking primary marrow disease. Flow analytes included CD45, SS, CD34, CD71, DRAQ 5 (DNA content), MPM-2 phosphorylation (mitoses), H2A.X phosphorylation (yH2A.X, phosphorylated by ATM in the presence of double strand DNA breaks), and Annexin V. Use of DRAQ 5 allowed analysis of DNA content in individual progenitor cell subsets (erythroid, myeloid, lymphoid, and stem cells); cycle results varied unexpectedly in individual subsets. Use of MPM-2 allowed confirmation of successful progression through the cell cycle (mitotic division), and simultaneous assessment of other parameters allowed this analysis in individual cell subsets. MDS patients demonstrated a significant increase in the G2 fraction in myeloid progenitors, with trends in S phase in myeloids and to a lesser extent post-G1 fractions in erythroid progenitors, with no proportionate increase in mitoses, yH2A.X binding was increased in erythroid and myeloid progenitors in MDS, with more modest elevation of Annexin V reactivity, yH2A.X and Annexin V reactivity varied with cell cycle compartment and lineage, and together were suggestive of DNA damage. Results suggest that MDS is characterized by DNA damage in hematopoietic progenitors, with post-G1 arrest or slowing of cell cycle progression, resulting in hypercellular marrow but failed marrow production of peripheral effector cells. These methods are useful for study of MDS biology. The abnormalities described may be useful as diagnostic tools.
M I N I M A L C R I T E R I A FOR DIAGNOSIS OF MDS
J. Bennett 1 *, G. Mufti 2. ICancer Center, University of Rochester, Rochester, NY, USA; 2Department of Hematology, King' s College, London; The MDS Foundation, Crosswicks, NJ, USA *E-mail: j [email protected] The W.H.O. has defined the minimal criteria for Dysplasia of cell lines as >10% dysplasia. However, the precise descriptions were lacking. An international morphology committee met on three occasions in 2005 and reviewed many cases of MDS. Criteria were established both for types of blasts; ringed sideroblasts; hypogranularity of neutrophils; megakaryoctic dysplasia (small nuclei and multiple nucleoli and megaloblastic erythroid forms. Each observer recorded the data electronically. In addition over 200 marrow cells were classified by each participant from the same images on a web site. The data is being analysed by a biostatistician for concordance and reproducibility and will be presented at the meeting. In addition plans are underway to organize a registry for Idiopathic Cytopenias of Unknown Significance (ICUS) for those patients who do not meet the minimal criteria.
[•-] CELL CYCLE STUDIES
IN M Y E L O D Y S P L A S T I C
SYN D ROME D.R. Head 1 *, J.W. Jacobberger 2, M. Jagasia 1, S. Goodman 1'3, L. Flye4, S. McClintock-Treep 1, E Boulos 1, E Jiang 1, G. Stelzer4, R. Briggs 1, K. Shults 4. 1Vanderbilt University Medical Center,
Nashville, TN, USA; 2Case Comprehensive Cancer Center, Cleveland, OH, USA; 3Nashville Veterans Administration
•
M Y E L O D Y S P L A S T I C S Y N D R O M E S IN C H I L D H O O D : EPIDEMIOLOGICAL ASPECTS
M. Hamaji 1 *, L.F. Lopes 1, M. do Rosario Latorre 2. 1Department
of Pediatrics, Hospital A. C. Camargo, S6o Paulo, Brazi# 2Department of Epidemiology, Universidade de S6o Paulo, S6o Paulo, Brazil *E-mail: [email protected] The data most commonly found on the etiology of the myelodysplastic syndromes (MDS) in children are based on genetic alterations, while there are few studies on exposure to toxic substances of patients and their parents. The possible etiology of environmental factors in MDS in childhood is unknown, and the influence of exposure to pesticides in adolescent and children has been argued. The main objective of this work was to take the personal and familial history of pediatric patients with MDS and compare them with data of children with acute lymphocytic leukemia (ALL), children with acute myeloid leukemia (AML) and children with no neoplasia. A questionnaire was elaborated and applied through direct interview and, in rare cases, by telephone. 150 children were interviewed: 50 with no neoplasia, 40 MDS, 50 ALL and 10 AML. There was a statistically significant association between skin color and diagnosis (p =0.031, with 100% of children with AML being Caucasian against 67% of those with MDS), birthweight (p = 0.039, with highest prevalence of low birthweight in MDS), previously having lived in an agricultural zone (p=0.003, with highest prevalence in those with AML, 20%, against 0% in those with