DNA Methylation and Hydroxymethylation Profile of CD34 + -Enriched Cell Products Intended for Autologous CD34 + Cell Transplantation

Abstract

Epigenetic dysregulation has been shown to limit functional capacity of aging hematopoietic stem cells, which may contribute to impaired outcome of hematopoietic stem cell-based therapies. The aim of our study was to gain better insight into the epigenetic profile of CD34⁺-enriched cell products intended for autologous CD34⁺ cell transplantation in patients with cardiomyopathy. We found global DNA methylation content significantly higher in immunoselected CD34⁺ cells compared to leukocytes in leukapheresis products (2.33 ± 1.03% vs. 1.84 ± 0.86%, p = 0.04). Global DNA hydroxymethylation content did not differ between CD34⁺ cells and leukocytes (p = 0.30). By measuring methylation levels of 94 stem cell transcription factors on a ready-to-use array, we identified 15 factors in which average promoter methylation was significantly different between leukocytes and CD34⁺ cells. The difference was highest for HOXC12 (58.18 ± 6.47% vs. 13.34 ± 24.18%, p = 0.0009) and NR2F2 (51.65 ± 25.89% vs. 7.66 ± 21.43%, p = 0.0045) genes. Our findings suggest that global DNA methylation and hydroxymethylation patterns as well as target methylation profile of selected genes in CD34⁺-enriched cell products do not differ significantly compared to leukapheresis products and, thus, can tell us little about the functional capacity and regenerative properties of CD34⁺ cells. Future studies should examine other CD34⁺ cell graft characteristics, which may serve as prognostic tools for autologous CD34⁺ cell transplantation.