Application of interphase fluorescence in situ hybridisation for the analysis of CD34+ cells in peripheral blood of patients with myelodysplastic syndromes
Abstract
Classical cytogenetic analysis of bone marrow cells is the standard for evaluating chromosomal aberrations in patients with myelodysplastic syndromes (MDS). However, there are several factors affecting the success of this method, such as the degree of cell proliferation, quality of preparations or size of pathological clone.
In these cases, it is possible to use interphase fluorescence in situ hybridization (I-FISH), which allows detection of chromosomal aberrations in the nuclei of interphase cells. In this study, we used the I-FISH method to screen chromosomal aberrations in peripheral blood circulating CD34+ cells.
Our objective was to evaluate the frequency of chromosomal aberrations in CD34+ cells isolated from peripheral blood, to compare this with the results of I-FISH analysis of bone marrow cells and to assess the significance of this method for monitoring the effect of therapy in patients with MDS. We examined 22 samples of CD34+ cells separated from the peripheral blood of 16 patients with MDS (13 females, 3 males; 11 x 5q- syndromes, 4x MDS RCMD and 1x MDS/MPS).
Four patients were investigated repeatedly. I-FISH on the separated CD34+ cells confirmed the prior findings in bone marrow in all the patients examined - 14 patients had deletion of chromosome 5 in the 5q31 region, one patient had deletion of chromosome 20q11q12 and one patient had monosomy of chromosome 7.
In our work, we demonstrated that isolation of CD34+ cells followed by I-FISH analysis is a practical, non-invasive method that can detect specific chromosomal abnormalities present in the pathological clone. This method has a predictive value and can be used for continuous disease control as well as for monitoring the pathological clone dynamics following treatment.