Application Of MLPA In Detection Of Chromosome Abnormalities Of Myelodysplastic Syndromes Long-term Outcome Of Thalidomide And Cyclosporine InPatients With IPSS Low/intermediate-1 MyelodysplasticSyndrome

Background and objectiveCytogenetic analysis provides important diagnostic and prognostic information for patients with myelodysplastic syndromes (MDS). Multiplex ligation-dependent probe amplification (MLPA) is a method that can detect copy number changes of up to 50 nucleic acid sequences in one simple reaction. MLPA quickly identifies amplifications and deletions of a large number of genes, and can be used for DNA, mRNA and CpG-methylation profiling of both blood and tumor-derived samples. In this study we describe the establishment of an innovative MDS MLPA assay for use in the detection of chromosomal abnormalities, in direct comparison to G or R-band karyotyping.MethodsIn the present study, MLPA analysis was applied to analyze cytogenetics of bone marrow cells of 437 MDS samples, and its result was compared with G or R-band karyotyping data. We used peripheral blood samples from 20 random normal volunteers to establish reference ranges. Kaplan-Meier method, Log-rank test and COX regression model were used to evaluate factors that influence the prognosis.Results1.437 MDS samples subjected to MLPA analysis passed the DNA quality control and were available for MLPA analysis. Overall,153 cases (35.0%) harbored at least one CNV. 2. Using MLPA analysis, clonal cytogenetic abnormalities were detected in 2/5 (40%) of MDS cases with karyotype failure and in 20/235 (8.5%) with normal G or R-band karyotypes, which may alter the risk stratification.3. Of the cases with abnormal G or R-band karyotypes,66/197 (33.5%) showed discrepancies between MLPA and G or R-band results. Among the discrepancy,12 cases had complex karyotypes.41 cases was attributed to a failure of MLPA probes in targeting the chromosomal abnormalities and 10 cases harbored small clones.4. In multivariate analyses, age (P=0.002, HR=1.021,95%CI 1.008-1.034),-7/7q-(P=0.040, HR=1.714,95%CI 1.025-2.865) and 17p-(P=0.018, HR=2.238,95%CI 1.148-4.363) were unfavorable prognostic factors independent of IPSS-R.ConclusionAs a trustworthy and efficient method, the high-resolution MPLA assay could be used in conjunction with G or R-band karyotping for the detection of chromosomal abnormalities in patients with suspected myelodysplastic syndromes, and it may provide more accuracy prognostic information.ObjectiveTo investigate the long-term outcome of Cyclosporin A combined with thalidomide for Chinese patients with IPSS low/intermediate-1 myelodysplastic syndromes without del(5q) and the predictive variables which could impact the response to the therapy.MethodsWe retrospectively analyzed seventy-six MDS patients who were treated with these drugs at a single institute in China. In order to confirm the predictive value of rsl672753 polymorphism of Cereblon, we detected it in sixty-three patients of our cohort.Results1. A total of 53% of patients showed hematological response to the therapy. Thirty-one patients (31/73,43%) achieved HI-E; fifteen patients (15/50,30%) achieved HI-N; eighteen patients (18/58,31%) achieved HI-P. Twenty-seven patients (46%) who were dependent on red blood cell transfusion achieved HI-E and independence from transfusion.2. The median duration of response among the responders was 22 months (range, 1-131+months) at the time of analysis. Bone marrow blasts ≤2% was the only variable tested associated with longer response duration in univariate analysis (P=0.010).3. We could not find any significant difference between the two groups of celeblon rsl672753 polymorphism either on the response rate or the response duration.4. The median survival of 67 patients without stem cell transplantation is 74 months. In multivariate analyses variables significantly correlated with survival were IPSS-R (HR=3.461,95%CI 1.126-10.639, P=0.030), age≥60y (HR=4.120,95%CI 1.070-15.867, P=0.040) and HI-N(HR=7.733,95%CI 1.007-59.396, P=0.049).ConclusionIn this study, we demonstrated the effectiveness and security of CsA combined with thalidomide in the treatment of IPSS low/intermediate-1 risk MDS patients without del(5q). The predictive value of cereblon gene polymorphism, rsl672753, could not be verified in this study.