The quality of the bone marrow sample for successful conventional cytogenetic analysis is important

Abstract In childhood acute lymphocyte leukemia (ALL), cytogenetics plays an important role in diagnosis, allocation of treatment and prognosis. On base of the conventional cytogenetic analysis, molecular methods have inproved our ability to accurately and rapidly risk-stratify patient with childhood ALL in the last few years. Our aim was to assess the demography of cytogenetic abnormalities in childhood ALL. The study sample consisted of 124 newly diagnosed ALL patients younger than 16 years, who were diagnosed at the Department of Pediatric Hematology/Oncology, Soochow University Children’s Hospital. The diagnosis and FAB subtypes of ALL was determined by Wright-Giemsa-stained bone marrow smears and cytochemicalstaining. Immunophenotyping of the bone marrow samples was performed by flow cytometry. Multiplex reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed to detect the 29 most common leukemia translocations for routine molecular diagnostic hematopathology practice, and complement the information gained from conventional cytogenetic analysis. Cytogenetic analysis was successful in 112 of 124 children with ALL. Sixty-eight (60%)of them had clonal chromosomal abnormalities. Numerical imbalances consisted of hyperdipoild(>47 chromosomes, 36 cases), hypodipoild(<46 chromosomes, 14 cases), pseudodiploidy(18 cases). Chromosomal translocations were observed in 13 patients by conventional cytogenetic analysis. Three cases were found positive for t (4;11), 3 cases for t (9;22), 1 case for t (1;19) and 6 cases for other rare translocations. RT-PCR analysis detected 116 of the 124 ALL patients. Thirteen cases of TEL-AML1, 10 cases of rearrangement in the MLL gene, 4 cases of E2A-PBX1, 4 cases of E2A-HLF, 3 cases of BCR-ABL, 2 cases of TLS-ERG, 32 cases of HOX11, were detected by RT-PCR in B-lineage leukemias. SIL-TAL1 had been found in 4 of 7 of T-lineage leukemias. Sixty-eight cases of ALL show chromosomal aberrations. Multiplex PCR positivity was detected in 59(50%)of the 116 ALL patients studied. Multiplex PCR combined with chromosome analysis uncovered Chromosomal abnormalities in 95 of 124(77%) of ALL patients and supplemented each other in detecting Chromosomal abnormalities. It provides reliable evidence for the diagnosis, classification and prognosis.

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Complex Karyotypic Abnormalities Detected by Conventional Cytogenetic Analysis More Strongly Predict Survival Than FISH, ZAP70, or IgVH Mutation Status for Previously Treated Patients with CLL.

Blood ◽

10.1182/blood.v110.11.2065.2065 ◽

2007 ◽

Vol 110 (11) ◽

pp. 2065-2065

Author(s):

William G. Wierda ◽

S. O’Brien ◽

S. Faderl ◽

A. Ferrajoli ◽

G. Garcia-Manero ◽

...

Keyword(s):

Bone Marrow ◽

Prognostic Factors ◽

High Risk ◽

Cytogenetic Analysis ◽

Independent Predictor ◽

Alkylating Agents ◽

Mutation Status ◽

Conventional Cytogenetic Analysis ◽

Previously Treated Patients ◽

Previously Treated

Abstract Recently, several novel prognostic factors have been identified; their significance has been demonstrated in selected patient (pt) populations and retrospective analyses. As a group, previously treated pts with CLL likely have their respective, relevant prognostic factors for clinical endpoints, which may be further impacted by treatment (Rx). We prospectively evaluated the significance of newer prognostic factors: FISH abnormalities (abn) (Vysis CLL panel), IgVH mutation status, ZAP70 expression (flow & immunohistochemistry), CD38 expression (≥30%); as well as traditional factors: conventional cytogenetic analysis perfomed on bone marrow metaphases, age, sex, # prior Rx, refractoriness to alkylating agents (ALK) or fludarabine (FLU), absolute lymphocyte count (ALC), HGB, PLT, β-2 microglobulin (B2M), ALB, LDH, creatinine, and Alk Phos as independent predictors for survival in previously treated pts. The group included 473 previously treated pts seen at M.D.Anderson (10/03–8/07), who were evaluated by bone marrow sampling with conventional and FISH cytogenetic analyses, and the new and traditional prognostic factors described above. The median (range) age was 63yrs(31–87) and # prior Rx was 2(1–13). Other characteristics were: 43% Rai high-risk; 35% FLU-refractory; and 39% ALK-refractory; 74% unmutated IgVH; 54% ZAP70+ (flow); 76% ZAP70+ (IHC); and 68% CD38+. FISH results were: 22% del 17p13, 21% del 11q22, 10% +12, and 48% del 13q14 or no abn by the hierarchical classification. Conventional cytogenetic analysis of bone marrow metaphases demonstrated 25% with a complex karyotypic abn (>1 cell with >1 chromosome abn), 58% diploid, 17% with single clonal abn (>1 cell with 1 abn). Of the 100 pts with complex karyotypic abn, 50% had del 17p13, 28% del 11q22, 6% +12, 9% del 13q14, and 7% had no abn by FISH. Survival was measured from the time of prognostic factor characterization (FISH). The median follow-up time was 10mo(0–47). Univariate analyses identified the following significant (p≤.01) predictors for shorter survival: advanced age, # prior Rx, Rai high-risk, ALK- or FLU-refractory, FISH del 17p13; complex karyotypic abn (Figure 1), unmutated IgVH, high ALC, low HGB, low PLT, high B2M, low ALB, high LDH, and high Alk Phos. Multivariate analysis produced the following model with the following significant (p<.05) independent predictors for survival: ALK- (HR 2.2) or FLU-refractory (1.9), complex karyotypic abn (HR 1.8), PLT (HR 0.99), and ALB (HR 0.35). We previously reported complex karyotypic abn as a significant independent predictor for shorter survival in previously treated patients receiving chemoimmunotherapy (JCO23:4070, 2005). These data indicate that for previously treated pts with CLL, a complex karyotypic abn detected by conventional cytogenetic analysis is a strong independent predictor for survival and appears superior to FISH, and other newer prognostic factors such as IgVH mutation status and ZAP70 expression. Figure Figure

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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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Application of Thrombolytic Drugs on Clotted Blood and Bone Marrow Specimens to Generate Usable Cells for Cytogenetic Analyses

Archives of Pathology & Laboratory Medicine ◽

10.5858/2010-0085-oar1.1 ◽

2011 ◽

Vol 135 (7) ◽

pp. 915-919

Author(s):

Angelique St Antoine ◽

Morgan N. Ketterling, BA ◽

William R. Sukov ◽

Josh Lowman ◽

Ryan A. Knudson ◽

...

Keyword(s):

Bone Marrow ◽

Mitotic Index ◽

Cytogenetic Analysis ◽

Chromosome Length ◽

Clinical Implementation ◽

Intact Cells ◽

Blood Samples ◽

Cytogenetic Analyses

Abstract Context.—Clotted blood and bone marrow specimens account for a large proportion of failed cytogenetic studies. There are no published protocols describing salvage of clotted specimens such that conventional chromosome or fluorescence in situ hybridization (FISH) studies can be performed. Objective.—To evaluate the utility of thrombolytic drugs on clotted blood samples to yield intact cells suitable for cytogenetic analysis. Design.—Five commercially available thrombolytic drugs (alteplase, urokinase, streptokinase, tenecteplase, reteplase) were evaluated in a series of blinded experiments to identify the best drug for lysing clots to produce samples suitable for chromosome and FISH studies. After the selection of alteplase as the drug yielding the most promising results, a comparative study between alteplase (0.75 mg/ml) and a commercially available anticlotting reagent (ACR) was performed. For each sample, mitotic index, chromosome length, and quality of slides prepared for conventional chromosome and FISH analyses were evaluated. Results.—Alteplase-treated samples produced a higher mitotic index than those treated with ACR while showing equivalent quality in conventional chromosome and FISH studies. We have demonstrated the utility of treating clotted blood samples with alteplase before cell culture to yield cells suitable for cytogenetic analysis. Since clinical implementation, this technique has been applied to more than 250 bone marrow samples, with a 93% success rate. Conclusions.—We believe the routine use of alteplase on clotted blood and bone marrow specimens should become standard for cytogenetics laboratories and may have similar utility in salvaging clotted specimens for other clinical assays requiring intact cells for analysis.

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The Utility of Peripheral Blood FISH in the Quantitation of BCR/ABL in CML Patients on Imatinib Mesylate: A Comparison with Bone Marrow FISH and Conventional Cytogenetics.

Blood ◽

10.1182/blood.v104.11.2942.2942 ◽

2004 ◽

Vol 104 (11) ◽

pp. 2942-2942

Author(s):

Samar Issa ◽

Duncan Holdsworth ◽

Paul Oei ◽

Peter J. Browett

Keyword(s):

Bone Marrow ◽

Imatinib Mesylate ◽

Peripheral Blood ◽

Cytogenetic Analysis ◽

Fusion Gene ◽

Chronic Phase ◽

Conventional Cytogenetic Analysis ◽

The Mean ◽

Conventional Cytogenetics ◽

Mean Differences

Abstract The hallmark of CML is the BCR/ABL fusion gene that is usually formed as a result of the t(9;22) translocation. Conventional cytogenetic analysis has been the standard method for monitoring the Philadelphia (Ph) chromosome, however evaluation of the BCR/ABL fusion gene using interphase Fluorescence in situHybridisation (FISH) on peripheral blood may allow more frequent and less invasive follow up of CML patients. The objective of this study was to compare the utility of peripheral blood FISH versus bone marrow FISH and conventional cytogenetics in patients with CML following treatment with Imatinib mesylate. 61 sets of peripheral blood and bone marrow aspirate samples from 33 Ph positive chronic phase CML patients receiving treatment with Imatinib mesylate were assessed from December 2002 to February 2004. Bone marrow samples were processed by standard cytogenetic procedures and G-banded analysis of at least 20 metaphases per sample was performed. Interphase FISH on non selected peripheral blood and bone marrow samples was carried out by scoring positive signals in 600 nuclei in each sample using BCR/ABL dual fusion or extra signal probes (Vysis). Bland and Altman plots were constructed to assess the level of agreement between the tests and the mean differences (with 95% confidence intervals) were determined. 13 of the 33 patients studied were male and 21 (64%) of the patients were analyzed at more than one timepoint. Although there was good agreement of peripheral blood FISH with bone marrow FISH and bone marrow cytogenetics in monitoring changes in the level of Ph positive cells following therapy, there were statistically significant differences in the agreement between the percentage levels of BCR/ABL positive cells between bone marrow cytogenetics and bone marrow and peripheral blood FISH. Cytogenetic analysis revealed significantly higher levels of BCR/ABL positive cells when compared to both peripheral blood FISH [9% (95% CI 4.6, 14.1), p=0.013] and bone marrow FISH [5% (95% CI 1.7, 7.8, p=0.013)]. The mean difference in the percentages of the Ph positive cells measured by bone marrow FISH exceeded the peripheral blood FISH by 5% (95% CI 1.0, 8.1, p=0.037). There was a significant relationship between the differences observed and the actual percentage level of BCR/ABL positive cells (P<0.001) with most of the discrepancy being driven by tests with BCR/ABL positive cells levels between 10 and 90%. In this subgroup, the cytogenetic tests were significantly higher [mean level 34% (95% CI 26, 45), p<0.001] than peripheral blood FISH and 23% (95% CI 13, 33, p<0.001) higher than bone marrow FISH. These observed differences may relate to the analysis of non-dividing cells in the FISH studies, including the assessment of Ph negative T lymphocytes in the peripheral blood, and need to be considered when monitoring patients by peripheral blood FISH studies alone. Figure Figure

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Clonal Cells Detected by Conventional Cytogenetic Analysis Correlates with Bone Marrow Blast Percentage and Progression Free Survival Into Acute Leukemia In Myelodysplastic Syndromes

Blood ◽

10.1182/blood.v116.21.2935.2935 ◽

2010 ◽

Vol 116 (21) ◽

pp. 2935-2935

Author(s):

Miyoung Kim ◽

Cha Ja She ◽

Sang Mee Hwang ◽

Sung-Soo Yoon ◽

Byoung Kook Kim ◽

...

Keyword(s):

Bone Marrow ◽

Prognostic Factors ◽

Cytogenetic Analysis ◽

Chromosomal Abnormalities ◽

Progression Free Survival ◽

Bone Marrow Blast ◽

Free Survival ◽

Conventional Cytogenetic Analysis ◽

Cell Percentage ◽

Clonal Abnormalities

Abstract Abstract 2935 Introduction: Chromosomal abnormalities not only demonstrate the clonality of the disease, but also help to predict the outcome of the patients in MDS. While IPSS suggests conventional cytogenetic analysis as a standard method in detecting clonal abnormalities in MDS, FISH is widely used due to its ability to detect minor clones and to quantitate the clonal abnormalities. In this study, we compared the results of conventional cytogenetic analysis and FISH, and investigated their different prognostic impact in MDS. Materials and Methods: The study included 129 MDS patients composed of 10 RA, 5 RARS, 15 RCMD, 39 RAEB-1, 33 RAEB-2 and 5 MDS, U (WHO classification, 2001). A standard protocol was used to perform conventional cytogenetic analysis (G-banding). Interphase FISH was performed to detect any abnormalities of chromosomes 5, 7, 8, 20 and 1. The result was analyzed as per the manufacturer¡&hibar;s instructions and recorded according to ISCN (2005) criteria. Result: The incidence of −5/5q, −7/7q, +8, −20q and +1q were 13.2%, 14.0%, 19.4%, 7.0% and 7.8%, respectively. Among them, FISH detected occult abnormalities which were not detected in conventional cytogenetic analysis in 1.6%, 3.1%, 5.4%, 1.6% and 5.4%, respectively. Overall, FISH detected occult abnormalities in 18.6% of the patients (24/129), and IPSS grouping was changed in 4.9% (6/129: 1 Low to Int-1; 4 Int-1 to Int-2; 1 Int-2 to High). The abnormalities in −5/5q, −7/7q and +1q were more common with other chromosomal abnormalities, whereas the abnormalities in +8 and −20q were more common as a sole abnormality. The quantity of clonal cells for each chromosome detected in conventional cytogenetic analysis did not correlate with the quantity of clonal cells detected in FISH. The presence of clonal cells either in conventional cytogenetic analysis or in FISH did not correlate with prognostic factors included in IPSS. However, the clonal cell percentage detected in conventional cytogenetic analysis was higher in patients with >5% bone marrow blasts than those with <5% (79.1% vs. 57.9%, p=.013). The clonal cell percentage detected in FISH did not show any relationship with IPSS factors including bone marrow blast percentage. Multivariate analysis showed the presence of clonal cells in conventional cytogenetic analysis were independent prognostic factors in predicting progression free survival into acute leukemia in this patient group (p=.038). Conclusion: FISH is advantageous in identifying cryptic cytogenetic abnormalities which could be overlooked in conventional cytogenetic analysis, and can change the IPSS risk grouping in MDS patients. The quantity of clonal cells detected in conventional cytogenetic analysis correlates with bone marrow blast percentage, whereas the quantity of clonal cells detected in FISH does not. Blasts possess higher proliferating activity than maturing hematopoietic precursors, thus they are more likely to form metaphase required in conventional cytognetic analysis. In contrast, the FISH results performed on non-dividing, interphase cells might reflect the true quantity of clonality both in blasts and maturing hematopoietic precursors. The relationship between the quantity of clonal cells and bone marrow blast percentage, and the prognostic significance of the presence of clonality suggest the novel diagnostic utility of conventional cytogenetic analysis in MDS. Disclosures: No relevant conflicts of interest to declare.

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Retrospective evaluation of chromosome 1 anomalies in hematologic malignancies: A single center study

Genetics & Applications ◽

10.31383/ga.vol3iss3pp17-24 ◽

2019 ◽

Vol 3 (3) ◽

pp. 17

Author(s):

Süreyya Bozkurt ◽

Müfide Okay ◽

Filiz Sağlam ◽

İbrahim C Haznedaroğlu

Keyword(s):

Bone Marrow ◽

Cytogenetic Analysis ◽

Hematologic Malignancies ◽

Chromosome 1 ◽

Chromosomal Anomalies ◽

Image Analysis System ◽

Conventional Cytogenetic Analysis ◽

Single Center Study ◽

Genetic Aberration ◽

Analysis System

Various anomalies of chromosome 1 which is the biggest chromosome among all human chromosomes were found in various hematologic diseases. In this retrospective study, clinical features and cytogenetic anomalies of 35 hematological patients with various chromosome 1 anomalies were correlated. Also the effect of chromosome 1 anomalies to the prognosis of those patients was discussed. Conventional cytogenetic analysis of those patients was performed by investigating metaphases of 24 hours stimulated bone marrow samples. After cell culturing, the samples were treated with tripsin and stained with Giemsa (GTG Banding). Analyses were performed on image analysis system (Metasystem, Germany). Chromosome 1 anomalies were determined in 35 patients (0.5 %) among 6865 samples having done their conventional bone marrow cytogenetic analysis in our center between January 2008 and March 2016. The ratio of chromosome 1 anomalies of totally 701 anomalies among 6865 patients was 4.9 %. Chromosome 1 anomalies were found mostly in patients with MM, MDS and AML in our study group. The most common anomaly was deletion 1 which was seen in 16 (37%) of the patients. Second most common anomaly was derivation 1 which was seen in 13 (30%) of the patients. Also translocations of chromosome 1 with other chromosomes were seen. The genetic aberration formed as a result of chromosomal anomalies result in the formation of hematologic malignancies. The effect on disease pathogenesis and prognosis of some of those anomalies is known and some needs to be investigated and determined.

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SNP/CGH Microarray Analysis in MDS: Correlation with Conventional Cytogenetic, FISH and Flow Cytometry Findings

Blood ◽

10.1182/blood.v124.21.5592.5592 ◽

2014 ◽

Vol 124 (21) ◽

pp. 5592-5592

Author(s):

Barbara Katharina Zehentner ◽

Lisa Eidenschink Brodersen ◽

Christine F Stephenson ◽

Jevon Cutler ◽

Monica E. de Baca ◽

...

Keyword(s):

Bone Marrow ◽

Flow Cytometry ◽

Microarray Analysis ◽

Cytogenetic Analysis ◽

Fish Analysis ◽

Employment Equity ◽

Conventional Cytogenetic Analysis ◽

Flow Cytometric ◽

Equity Ownership ◽

Gains And Losses

Abstract Background: Single nucleotide polymorphism (SNP) and comparative genomic hybridization (CGH) microarray analysis is a powerful tool to assess myelodysplastic bone marrow specimens for the presence of genomic gains and losses as well as loss of heterozygosity (LOH) (reviewed by Nybakken & Bagg, JMD 2014). Its application can be a valuable addition to conventional cytogenetic analysis and may be superior to FISH testing for MDS assessment. Currently, microarray analysis does not have widespread use in an MDS work-up. Several groups have demonstrated that flow cytometric analysis can detect phenotypic aberrations in bone marrow aspirates with cytopenias with more abnormalities identified in patients with poor prognosis or with multiple genotypic abnormalities (Loken et al. 2008; Cutler et al. 2011; van de Loosdrecht et al. 2013). In this study SNP microarray results were compared with conventional cytogenetic and MDS panel FISH findings as well as phenotypic abnormalities detected by flow cytometry. Patients and Methods: 185 bone marrow aspirate specimens submitted to our laboratory for MDS work-up were analyzed by SNP/CGH studies. 36 of these (19.5%) were positive by SNP/CGH microarray analysis. 32 of the positive microarray cases (88.9%) were also analyzed by conventional cytogenetic studies, 35 (97.2%) by MDS FISH panel (5p, 7q, +8, -17p, -20q) and 31 (86.1%) were assessed by multidimensional flow cytometry (FCM) and were assigned an FCSS score (Wells et al. 2003). Results: Of the specimens in which the SNP/CGH array demonstrated genotypic abnormalities, 11/32 (34.4%) were negative by conventional cytogenetic analysis while 12/35 (34.3%) showed no abnormalities by MDS FISH panel analysis. SNP/CGH analysis revealed additional chromosomal gains and losses in 18/32 (56%) in comparison to cytogenetic analysis and in 22/35 (63%) in comparison to FISH analysis. Loss of Heterozygosity regions were detected in 28/36 cases (78%) with 96.4% (27/28) of these being larger than 2 Mb and 53% (19/28) spanning a significant chromosomal region (e.g. 1p, 5q, 7q and 17p) with known oncogenic and other MDS related genes. In 10/32 cases (31%), microarray analysis was able to characterize the origin of marker chromosome material, previously reported with unknown identity by conventional cytogenetic analysis. In an additional subset of 10 out of 32 cases (31%), cytogenetic analysis was able to either characterize balanced translocations or low level sub-clonal abnormalities not identified by microarray analysis alone. In 11/36 (31%) microarray analysis was able to detect clonal heterogeneity and evolution. In none of the specimens did FISH analysis detected abnormalities not revealed by microarray analysis. Flow cytometry performed on 31 of the array positive specimens revealed 6 to have >20% abnormal myeloid progenitor cells (classified as AML) while 23 the remaining 25 cases showed phenotypic abnormalities consistent with MDS (FCSS ranging from 1-6). In two specimens with a FCSS of 0, LOH regions on 16q or 1p and 21q were found, respectively, without the presence of numerical aberrations. A FCSS score of 1 with minimal phenotypic abnormalities (n=3), was comprised of one specimen with del(5q), one with LOH of 7q and one with trisomy 8, 1p loss and 1q gain. Specimens with an FCSS of 2 (n=7) showed only one specimen classified as complex (5 or more abnormalities). The two FCSS =3 specimens showed del(5q) with del(12p) and several LOH regions, not complex findings. One of the 4 specimen with FCSS = 4 was classified as complex while the other 3 specimens showed monosomy 7, LOH of 7q or LOH of 1p, respectively. Genotypic abnormalities were also related to phenotypic abnormalities in 4/7 (57%) specimens in the FCSS = 5/6 category which revealed complex microarray findings. Half (3/6) of the AML class had complex findings as well. Conclusions: These results emphasize the additional value that CGH/SNP microarray analysis adds to conventional cytogenetic analysis. Our dataset confirms that FISH studies do not provide additional information for MDS specimens positive by cytogenetic and/or microarray analysis. Most importantly, a high correlation between our phenotypic flow cytometric scoring system for myeloid abnormalities and microarray findings has been identified. Higher flow cytometric abnormality scores correlate with increasing complexity of genomic abnormalities. Disclosures Zehentner: HematoLogics Inc.: Employment, Equity Ownership. Brodersen:Hematologics Inc.: Employment. Stephenson:Hematologics Inc.: Employment. de Baca:Hematologics Inc.: Employment. Menssen:Hematologics Inc.: Employment. Hammock:Hematologics Inc.: Employment. Johnson:Hematologics Inc.: Employment. Hartmann:Hematologics Inc.: Employment. Loken:Hematologics Inc.: Employment, Equity Ownership. Wells:HematoLogics Inc.: Employment, Equity Ownership.

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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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