Isolation, Characterization and Application of BCR-ABL-ve Mesenchymal Stem Cells Derived from Peripheral Blood of Chronic Myeloid Leukemia (CML) in Enhancing Potentiality of Bone Marrow Transplantation in CML Patients

Abstract

Chronic Myeloid leukemia (CML) is a disorder causing uncontrolled growth of myeloproliferative blast cells. Tyrosine kinase inhibitors are not really useful in advanced stages and have their own side effects, reactions and resistance as well. Till date, CML can be fully cured only by bone marrow transplantation (BMT) which has its own limitations and thus need further studies to increase better survival of CML patients by this procedure. Our lab at Jaslok Hospital has been working on studying and characterizing mesenchymal stem cells in various types of hematological malignancies. We came up with a concept of isolating and characterizing mesenchymal stem cells (MSCs) from peripheral blood of a CML patient which can be used in combination with Bone Marrow Transplantation in the same CML patient. Our study has shown that the stem cells derived from peripheral blood of CML patient were found to be BCR/ABL -ve . We designated this cell line as a "BCR/ABL -ve " cell line. Molecular characterization of these cells further confirmed their Mesenchymal phenotypes with distinct expression of CD105, CD13 and CD73 genes. Interestingly these cells also expressed pluripotency genes such as OCT4 and NANOG and cytokines i.e. IL6 and TNFα. We further confirmed the normal phenotype of BCR/ABL -ve MSCs by localizing expression of H-Ras, Rb, P53, P16, P21, and EGFR and Ki67 cancer related proteins in these cells by immunofluorescence Microscopy and by in vitro transformation assay. Thus, we suggest that these normal BCR/ABL -ve MSCs derived from CML patient's peripheral blood can be used in addition to BMT procedure for a better recovery of CML disease in near future.

Loss of death-associated protein kinase 1 in human bone marrow mesenchymal stem cells decreases immunosuppression of CD4+ T cells

“…A previous study used RT-PCR to demonstrate that MSCs from the peripheral blood of acute lymphoblastic leukemia patients prominently expressed the DAPK1 gene. 27 Our results demonstrated high DAPK1 expression at the protein level in ex vivo-expanded hMSCs isolated from bone marrow (Figure 2a and 2b) and documented a correlation between DAPK1 expression level and MSC immunosuppression of CD4þ T cells (Figure 2d and 2e). These results prompted us to focus on the contribution of DAPK1 to the immunoregulatory functions of hMSCs.…”