CONVENTIONAL CYTOGENETICS, ACGH, AND PCR AS INTEGRATED WORKUP FOR A CORRECT DIAGNOSIS OF MDS.
(Abstract release date: 05/19/16)
EHA Library. Galimberti S. 06/09/16; 132752; E1203
Prof. Sara Galimberti
Contributions
Contributions
Abstract
Abstract: E1203
Type: Eposter Presentation
Background
Conventional cytogenetics continues to have a fundamental role in the classification and risk scoring of myelodysplastic syndromes (MDS). Nevertheless, non-informative karyotypes represent up to 20% of cases. Some different molecular methods, such as FISH, aCGH or mutational analysis, could detect new abnormalities and improve the correct subtyping of MDS patients.
Aims
The aim of the study was to adopt an integrated diagnostic work-up for better characterizing MDS patients.
Methods
We analyzed 61 patients (71% male, with a median age of 74 years). According to the WHO classification, 33% were affected by multilineage refractory cytopenias, 12% by refractory anemia, 15% by refractory anemia with ring sideroblasts, 21% by refractory anemia with excess of blasts, 3% by 5q- syndrome, and 12% by chronic myelo-monocytic leukemia; 67% of patients were at low/intermediate-1 IPSS risk.All patients were assessed by conventional cytogenetics, FISH for chromosomes 5, 7, 8, PDGFRalpha and beta rearrangements, aCGH, and PCR for ASXL1, EZH2, TP53, and TET2 mutations. WT1 and RPS14 gene expression levels were also measured by quantitative PCR.
Results
In our series, conventional cytogenetics analysis failed in only 12% of cases: indeed, the sampling for this analysis was the first one during the bone marrow aspiration. Overall, 39% of patients showed at least one chromosomal aberration, with complex karyotypes in 7% of cases.FISH allowed to correctly classify two cases as affected by the 5q- syndrome and one as affected by deletion of chromosome 7; two patients carried PDGFRbeta rearrangement; these abnormalities had not been detected by the conventional cytogenetics.The aGCH allowed to detect chromosomal aberrations in 38% of cases: aCGH detected 10 “new” mutated cases in respect of the conventional cytogenetics, including alterations of the ETV6 and GATA2 genes;After the mutational analyses, 28% of patients resulted mutated, with highest frequency for TP53 (mutated in the 16% of the overall series). Eight of these TP53-mutated patients showed normal karyotype, and resulted wild-type by FISH and aCGH. Four low/intermediate-1 risk patients (8%) showed the ASXL1 gene mutation. Two cases showed the TET2 mutation.The statistical analysis confirmed the prognostic role of poor cytogenetics either on overall survival (OS) or progression-free-survival (PFS). Also deletions detected by aCGH resulted to play a negative prognostic impact on OS.WT1 and RPS14 gene expression was assessed: over-expression of WT1 was found in one third of all patients, while 70% showed RPS14 values lower than those measured in healthy donors. Statistical analysis showed that the WT1 over-expression had statistical significance on survival, also in multivariate analysis.
Conclusion
Our study supports the feasibility and the utility of the introduction in the routinary workup of MDS of FISH and aCGH, in order to better stratify MDS patients and correctly design ab initio a patient-tailored treatment.
Session topic: E-poster
Keyword(s): Array based comparative genomic hybridization, FISH, MDS, WT1
Type: Eposter Presentation
Background
Conventional cytogenetics continues to have a fundamental role in the classification and risk scoring of myelodysplastic syndromes (MDS). Nevertheless, non-informative karyotypes represent up to 20% of cases. Some different molecular methods, such as FISH, aCGH or mutational analysis, could detect new abnormalities and improve the correct subtyping of MDS patients.
Aims
The aim of the study was to adopt an integrated diagnostic work-up for better characterizing MDS patients.
Methods
We analyzed 61 patients (71% male, with a median age of 74 years). According to the WHO classification, 33% were affected by multilineage refractory cytopenias, 12% by refractory anemia, 15% by refractory anemia with ring sideroblasts, 21% by refractory anemia with excess of blasts, 3% by 5q- syndrome, and 12% by chronic myelo-monocytic leukemia; 67% of patients were at low/intermediate-1 IPSS risk.All patients were assessed by conventional cytogenetics, FISH for chromosomes 5, 7, 8, PDGFRalpha and beta rearrangements, aCGH, and PCR for ASXL1, EZH2, TP53, and TET2 mutations. WT1 and RPS14 gene expression levels were also measured by quantitative PCR.
Results
In our series, conventional cytogenetics analysis failed in only 12% of cases: indeed, the sampling for this analysis was the first one during the bone marrow aspiration. Overall, 39% of patients showed at least one chromosomal aberration, with complex karyotypes in 7% of cases.FISH allowed to correctly classify two cases as affected by the 5q- syndrome and one as affected by deletion of chromosome 7; two patients carried PDGFRbeta rearrangement; these abnormalities had not been detected by the conventional cytogenetics.The aGCH allowed to detect chromosomal aberrations in 38% of cases: aCGH detected 10 “new” mutated cases in respect of the conventional cytogenetics, including alterations of the ETV6 and GATA2 genes;After the mutational analyses, 28% of patients resulted mutated, with highest frequency for TP53 (mutated in the 16% of the overall series). Eight of these TP53-mutated patients showed normal karyotype, and resulted wild-type by FISH and aCGH. Four low/intermediate-1 risk patients (8%) showed the ASXL1 gene mutation. Two cases showed the TET2 mutation.The statistical analysis confirmed the prognostic role of poor cytogenetics either on overall survival (OS) or progression-free-survival (PFS). Also deletions detected by aCGH resulted to play a negative prognostic impact on OS.WT1 and RPS14 gene expression was assessed: over-expression of WT1 was found in one third of all patients, while 70% showed RPS14 values lower than those measured in healthy donors. Statistical analysis showed that the WT1 over-expression had statistical significance on survival, also in multivariate analysis.
Conclusion
Our study supports the feasibility and the utility of the introduction in the routinary workup of MDS of FISH and aCGH, in order to better stratify MDS patients and correctly design ab initio a patient-tailored treatment.
Session topic: E-poster
Keyword(s): Array based comparative genomic hybridization, FISH, MDS, WT1
Abstract: E1203
Type: Eposter Presentation
Background
Conventional cytogenetics continues to have a fundamental role in the classification and risk scoring of myelodysplastic syndromes (MDS). Nevertheless, non-informative karyotypes represent up to 20% of cases. Some different molecular methods, such as FISH, aCGH or mutational analysis, could detect new abnormalities and improve the correct subtyping of MDS patients.
Aims
The aim of the study was to adopt an integrated diagnostic work-up for better characterizing MDS patients.
Methods
We analyzed 61 patients (71% male, with a median age of 74 years). According to the WHO classification, 33% were affected by multilineage refractory cytopenias, 12% by refractory anemia, 15% by refractory anemia with ring sideroblasts, 21% by refractory anemia with excess of blasts, 3% by 5q- syndrome, and 12% by chronic myelo-monocytic leukemia; 67% of patients were at low/intermediate-1 IPSS risk.All patients were assessed by conventional cytogenetics, FISH for chromosomes 5, 7, 8, PDGFRalpha and beta rearrangements, aCGH, and PCR for ASXL1, EZH2, TP53, and TET2 mutations. WT1 and RPS14 gene expression levels were also measured by quantitative PCR.
Results
In our series, conventional cytogenetics analysis failed in only 12% of cases: indeed, the sampling for this analysis was the first one during the bone marrow aspiration. Overall, 39% of patients showed at least one chromosomal aberration, with complex karyotypes in 7% of cases.FISH allowed to correctly classify two cases as affected by the 5q- syndrome and one as affected by deletion of chromosome 7; two patients carried PDGFRbeta rearrangement; these abnormalities had not been detected by the conventional cytogenetics.The aGCH allowed to detect chromosomal aberrations in 38% of cases: aCGH detected 10 “new” mutated cases in respect of the conventional cytogenetics, including alterations of the ETV6 and GATA2 genes;After the mutational analyses, 28% of patients resulted mutated, with highest frequency for TP53 (mutated in the 16% of the overall series). Eight of these TP53-mutated patients showed normal karyotype, and resulted wild-type by FISH and aCGH. Four low/intermediate-1 risk patients (8%) showed the ASXL1 gene mutation. Two cases showed the TET2 mutation.The statistical analysis confirmed the prognostic role of poor cytogenetics either on overall survival (OS) or progression-free-survival (PFS). Also deletions detected by aCGH resulted to play a negative prognostic impact on OS.WT1 and RPS14 gene expression was assessed: over-expression of WT1 was found in one third of all patients, while 70% showed RPS14 values lower than those measured in healthy donors. Statistical analysis showed that the WT1 over-expression had statistical significance on survival, also in multivariate analysis.
Conclusion
Our study supports the feasibility and the utility of the introduction in the routinary workup of MDS of FISH and aCGH, in order to better stratify MDS patients and correctly design ab initio a patient-tailored treatment.
Session topic: E-poster
Keyword(s): Array based comparative genomic hybridization, FISH, MDS, WT1
Type: Eposter Presentation
Background
Conventional cytogenetics continues to have a fundamental role in the classification and risk scoring of myelodysplastic syndromes (MDS). Nevertheless, non-informative karyotypes represent up to 20% of cases. Some different molecular methods, such as FISH, aCGH or mutational analysis, could detect new abnormalities and improve the correct subtyping of MDS patients.
Aims
The aim of the study was to adopt an integrated diagnostic work-up for better characterizing MDS patients.
Methods
We analyzed 61 patients (71% male, with a median age of 74 years). According to the WHO classification, 33% were affected by multilineage refractory cytopenias, 12% by refractory anemia, 15% by refractory anemia with ring sideroblasts, 21% by refractory anemia with excess of blasts, 3% by 5q- syndrome, and 12% by chronic myelo-monocytic leukemia; 67% of patients were at low/intermediate-1 IPSS risk.All patients were assessed by conventional cytogenetics, FISH for chromosomes 5, 7, 8, PDGFRalpha and beta rearrangements, aCGH, and PCR for ASXL1, EZH2, TP53, and TET2 mutations. WT1 and RPS14 gene expression levels were also measured by quantitative PCR.
Results
In our series, conventional cytogenetics analysis failed in only 12% of cases: indeed, the sampling for this analysis was the first one during the bone marrow aspiration. Overall, 39% of patients showed at least one chromosomal aberration, with complex karyotypes in 7% of cases.FISH allowed to correctly classify two cases as affected by the 5q- syndrome and one as affected by deletion of chromosome 7; two patients carried PDGFRbeta rearrangement; these abnormalities had not been detected by the conventional cytogenetics.The aGCH allowed to detect chromosomal aberrations in 38% of cases: aCGH detected 10 “new” mutated cases in respect of the conventional cytogenetics, including alterations of the ETV6 and GATA2 genes;After the mutational analyses, 28% of patients resulted mutated, with highest frequency for TP53 (mutated in the 16% of the overall series). Eight of these TP53-mutated patients showed normal karyotype, and resulted wild-type by FISH and aCGH. Four low/intermediate-1 risk patients (8%) showed the ASXL1 gene mutation. Two cases showed the TET2 mutation.The statistical analysis confirmed the prognostic role of poor cytogenetics either on overall survival (OS) or progression-free-survival (PFS). Also deletions detected by aCGH resulted to play a negative prognostic impact on OS.WT1 and RPS14 gene expression was assessed: over-expression of WT1 was found in one third of all patients, while 70% showed RPS14 values lower than those measured in healthy donors. Statistical analysis showed that the WT1 over-expression had statistical significance on survival, also in multivariate analysis.
Conclusion
Our study supports the feasibility and the utility of the introduction in the routinary workup of MDS of FISH and aCGH, in order to better stratify MDS patients and correctly design ab initio a patient-tailored treatment.
Session topic: E-poster
Keyword(s): Array based comparative genomic hybridization, FISH, MDS, WT1
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