DIFFERENT WHO SUBTYPES OF CHROMOSOMALLY NORMAL MDS SHARE COMMON AMPLIFIED AREAS ON ACGH/SNPA.
(Abstract release date: 05/19/16)
EHA Library. Bernasconi P. 06/09/16; 134803; PB1903
Prof. Dr. Paolo Bernasconi
Contributions
Contributions
Abstract
Abstract: PB1903
Type: Publication Only
Background
Various studies have reported that aCGH and SNPa can reveal hidden prognostically relevant karyotype alterations in chromosomally normal MDS patients as compared to conventional cytogenetics: aCGH offers a superior resolution of unbalanced chromosomal defects and SNPa identifies areas of copy neutral loss of heterozygosity (LOH).
Aims
Thus, the principal aims of the present study were to test the power of aCGH/SNPa in chromosomally normal MDS patients, to establish whether the extension and the number of cryptic gains/losses and LOH areas correlated with the WHO MDS subtype and to check whether patients with the same WHO subtype shared common abnormal areas.
Methods
aCGH/SNPa were carried out on the DNA of mononuclear cells from fifteen patients examined between January 2013 and December 2015. There were four females and eleven males, whose median age was 66 years (range 24-78). According to WHO classification, 9 patients were diagnose as RA, one as RA with ringed sideroblasts, one as refractory cytopenia with multilineage dysplasia (RCMD), one as RAEB-1 and 3 as RAEB-2. According to R-IPSS, 11 patients were considered low-risk, 2 intermediate-1 risk and 2 intermediate-2 risk. Median follow-up was eight months (range 1-23). At the time of the analyses one patient who progressed to AML and was submitted to two allogeneic HSCT (allo-HSCT) has relapsed and died; the RAEB-1 patient progressed to RAEB-2 and is alive nine months after an allo-HSCT. Molecular karyotyping was performed using the SurePrint G3 Human Cancer CGH+SNP Microarray Kit 4X180 (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s enzimatic labeling protocol. The array was scanned through the Agilent scanner G26000DA and analysed with CytoGenomics software (v3.0.6). FISH was carried out to confirm aCGH/SNPa results; commercial probes were obtained from Abbot Molecular Inc. (Chicago, Il, USA) and Kreatech (Amsterdam, NL) and applied according to manufacturer’s guidelines.
Results
aCGH/SNPa revealed an abnormal pattern in all MDS patients. In the three RAEB-2 patients aCGH showed the highest number of cryptic chromosomal lesions and many areas of LOH. Unexpectedly, two RAEB-2 patients presented a monosomy 7 together with additional defects including a p53 loss and a 5q deletion. The clinical value of these aCGH/SNPa results was strengthened by the fact that one patient progressed to AML that never achieved complete remission despite two allo-HSCT, whereas the other two patients experienced a quick AML progression. In the other twelve patients aCGH/SNPa detected small chromosomal lesions which size varied between 5kb and 1.8Mb and LOH areas of much longer size (range 1Mb-5.7Mb). Interestingly, by comparing the various chromosomal lesions, it was observed that different patients with different WHO MDS subtypes shared common amplified areas that included the NOTCH1 gene in five patients and the FGFR3 gene in three.
Conclusion
i) aCGH/SNPa can truly reveal cryptic chromosomal lesions in chromosomally normal MDS patients, a finding that determines a change in patients’ R-IPSS score; ii) advanced MDS presented the highest number of lesions which size was longer than that of lesions present in early MDS; iii) patients with different WHO MDS subtypes shared common amplified areas that contained cell cycle genes with a potential role in MDS pathogenesis.
Session topic: E-poster
Keyword(s): CGH, FISH, Myelodysplasia
Type: Publication Only
Background
Various studies have reported that aCGH and SNPa can reveal hidden prognostically relevant karyotype alterations in chromosomally normal MDS patients as compared to conventional cytogenetics: aCGH offers a superior resolution of unbalanced chromosomal defects and SNPa identifies areas of copy neutral loss of heterozygosity (LOH).
Aims
Thus, the principal aims of the present study were to test the power of aCGH/SNPa in chromosomally normal MDS patients, to establish whether the extension and the number of cryptic gains/losses and LOH areas correlated with the WHO MDS subtype and to check whether patients with the same WHO subtype shared common abnormal areas.
Methods
aCGH/SNPa were carried out on the DNA of mononuclear cells from fifteen patients examined between January 2013 and December 2015. There were four females and eleven males, whose median age was 66 years (range 24-78). According to WHO classification, 9 patients were diagnose as RA, one as RA with ringed sideroblasts, one as refractory cytopenia with multilineage dysplasia (RCMD), one as RAEB-1 and 3 as RAEB-2. According to R-IPSS, 11 patients were considered low-risk, 2 intermediate-1 risk and 2 intermediate-2 risk. Median follow-up was eight months (range 1-23). At the time of the analyses one patient who progressed to AML and was submitted to two allogeneic HSCT (allo-HSCT) has relapsed and died; the RAEB-1 patient progressed to RAEB-2 and is alive nine months after an allo-HSCT. Molecular karyotyping was performed using the SurePrint G3 Human Cancer CGH+SNP Microarray Kit 4X180 (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s enzimatic labeling protocol. The array was scanned through the Agilent scanner G26000DA and analysed with CytoGenomics software (v3.0.6). FISH was carried out to confirm aCGH/SNPa results; commercial probes were obtained from Abbot Molecular Inc. (Chicago, Il, USA) and Kreatech (Amsterdam, NL) and applied according to manufacturer’s guidelines.
Results
aCGH/SNPa revealed an abnormal pattern in all MDS patients. In the three RAEB-2 patients aCGH showed the highest number of cryptic chromosomal lesions and many areas of LOH. Unexpectedly, two RAEB-2 patients presented a monosomy 7 together with additional defects including a p53 loss and a 5q deletion. The clinical value of these aCGH/SNPa results was strengthened by the fact that one patient progressed to AML that never achieved complete remission despite two allo-HSCT, whereas the other two patients experienced a quick AML progression. In the other twelve patients aCGH/SNPa detected small chromosomal lesions which size varied between 5kb and 1.8Mb and LOH areas of much longer size (range 1Mb-5.7Mb). Interestingly, by comparing the various chromosomal lesions, it was observed that different patients with different WHO MDS subtypes shared common amplified areas that included the NOTCH1 gene in five patients and the FGFR3 gene in three.
Conclusion
i) aCGH/SNPa can truly reveal cryptic chromosomal lesions in chromosomally normal MDS patients, a finding that determines a change in patients’ R-IPSS score; ii) advanced MDS presented the highest number of lesions which size was longer than that of lesions present in early MDS; iii) patients with different WHO MDS subtypes shared common amplified areas that contained cell cycle genes with a potential role in MDS pathogenesis.
Session topic: E-poster
Keyword(s): CGH, FISH, Myelodysplasia
Abstract: PB1903
Type: Publication Only
Background
Various studies have reported that aCGH and SNPa can reveal hidden prognostically relevant karyotype alterations in chromosomally normal MDS patients as compared to conventional cytogenetics: aCGH offers a superior resolution of unbalanced chromosomal defects and SNPa identifies areas of copy neutral loss of heterozygosity (LOH).
Aims
Thus, the principal aims of the present study were to test the power of aCGH/SNPa in chromosomally normal MDS patients, to establish whether the extension and the number of cryptic gains/losses and LOH areas correlated with the WHO MDS subtype and to check whether patients with the same WHO subtype shared common abnormal areas.
Methods
aCGH/SNPa were carried out on the DNA of mononuclear cells from fifteen patients examined between January 2013 and December 2015. There were four females and eleven males, whose median age was 66 years (range 24-78). According to WHO classification, 9 patients were diagnose as RA, one as RA with ringed sideroblasts, one as refractory cytopenia with multilineage dysplasia (RCMD), one as RAEB-1 and 3 as RAEB-2. According to R-IPSS, 11 patients were considered low-risk, 2 intermediate-1 risk and 2 intermediate-2 risk. Median follow-up was eight months (range 1-23). At the time of the analyses one patient who progressed to AML and was submitted to two allogeneic HSCT (allo-HSCT) has relapsed and died; the RAEB-1 patient progressed to RAEB-2 and is alive nine months after an allo-HSCT. Molecular karyotyping was performed using the SurePrint G3 Human Cancer CGH+SNP Microarray Kit 4X180 (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s enzimatic labeling protocol. The array was scanned through the Agilent scanner G26000DA and analysed with CytoGenomics software (v3.0.6). FISH was carried out to confirm aCGH/SNPa results; commercial probes were obtained from Abbot Molecular Inc. (Chicago, Il, USA) and Kreatech (Amsterdam, NL) and applied according to manufacturer’s guidelines.
Results
aCGH/SNPa revealed an abnormal pattern in all MDS patients. In the three RAEB-2 patients aCGH showed the highest number of cryptic chromosomal lesions and many areas of LOH. Unexpectedly, two RAEB-2 patients presented a monosomy 7 together with additional defects including a p53 loss and a 5q deletion. The clinical value of these aCGH/SNPa results was strengthened by the fact that one patient progressed to AML that never achieved complete remission despite two allo-HSCT, whereas the other two patients experienced a quick AML progression. In the other twelve patients aCGH/SNPa detected small chromosomal lesions which size varied between 5kb and 1.8Mb and LOH areas of much longer size (range 1Mb-5.7Mb). Interestingly, by comparing the various chromosomal lesions, it was observed that different patients with different WHO MDS subtypes shared common amplified areas that included the NOTCH1 gene in five patients and the FGFR3 gene in three.
Conclusion
i) aCGH/SNPa can truly reveal cryptic chromosomal lesions in chromosomally normal MDS patients, a finding that determines a change in patients’ R-IPSS score; ii) advanced MDS presented the highest number of lesions which size was longer than that of lesions present in early MDS; iii) patients with different WHO MDS subtypes shared common amplified areas that contained cell cycle genes with a potential role in MDS pathogenesis.
Session topic: E-poster
Keyword(s): CGH, FISH, Myelodysplasia
Type: Publication Only
Background
Various studies have reported that aCGH and SNPa can reveal hidden prognostically relevant karyotype alterations in chromosomally normal MDS patients as compared to conventional cytogenetics: aCGH offers a superior resolution of unbalanced chromosomal defects and SNPa identifies areas of copy neutral loss of heterozygosity (LOH).
Aims
Thus, the principal aims of the present study were to test the power of aCGH/SNPa in chromosomally normal MDS patients, to establish whether the extension and the number of cryptic gains/losses and LOH areas correlated with the WHO MDS subtype and to check whether patients with the same WHO subtype shared common abnormal areas.
Methods
aCGH/SNPa were carried out on the DNA of mononuclear cells from fifteen patients examined between January 2013 and December 2015. There were four females and eleven males, whose median age was 66 years (range 24-78). According to WHO classification, 9 patients were diagnose as RA, one as RA with ringed sideroblasts, one as refractory cytopenia with multilineage dysplasia (RCMD), one as RAEB-1 and 3 as RAEB-2. According to R-IPSS, 11 patients were considered low-risk, 2 intermediate-1 risk and 2 intermediate-2 risk. Median follow-up was eight months (range 1-23). At the time of the analyses one patient who progressed to AML and was submitted to two allogeneic HSCT (allo-HSCT) has relapsed and died; the RAEB-1 patient progressed to RAEB-2 and is alive nine months after an allo-HSCT. Molecular karyotyping was performed using the SurePrint G3 Human Cancer CGH+SNP Microarray Kit 4X180 (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer’s enzimatic labeling protocol. The array was scanned through the Agilent scanner G26000DA and analysed with CytoGenomics software (v3.0.6). FISH was carried out to confirm aCGH/SNPa results; commercial probes were obtained from Abbot Molecular Inc. (Chicago, Il, USA) and Kreatech (Amsterdam, NL) and applied according to manufacturer’s guidelines.
Results
aCGH/SNPa revealed an abnormal pattern in all MDS patients. In the three RAEB-2 patients aCGH showed the highest number of cryptic chromosomal lesions and many areas of LOH. Unexpectedly, two RAEB-2 patients presented a monosomy 7 together with additional defects including a p53 loss and a 5q deletion. The clinical value of these aCGH/SNPa results was strengthened by the fact that one patient progressed to AML that never achieved complete remission despite two allo-HSCT, whereas the other two patients experienced a quick AML progression. In the other twelve patients aCGH/SNPa detected small chromosomal lesions which size varied between 5kb and 1.8Mb and LOH areas of much longer size (range 1Mb-5.7Mb). Interestingly, by comparing the various chromosomal lesions, it was observed that different patients with different WHO MDS subtypes shared common amplified areas that included the NOTCH1 gene in five patients and the FGFR3 gene in three.
Conclusion
i) aCGH/SNPa can truly reveal cryptic chromosomal lesions in chromosomally normal MDS patients, a finding that determines a change in patients’ R-IPSS score; ii) advanced MDS presented the highest number of lesions which size was longer than that of lesions present in early MDS; iii) patients with different WHO MDS subtypes shared common amplified areas that contained cell cycle genes with a potential role in MDS pathogenesis.
Session topic: E-poster
Keyword(s): CGH, FISH, Myelodysplasia
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