Histone Arginine Methylation-Mediated Epigenetic Regulation of Discoidin Domain Receptor 2 Controls the Senescence of Human Bone Marrow Mesenchymal Stem Cells

Abstract

The application of human bone marrow mesenchymal stem cells (hBM-MSCs) in cell-based clinical therapies is hindered by the limited number of cells remaining after the initial isolation process and by cellular senescence following in vitro expansion. Understanding the process of in vitro senescence in hBM-MSCs would enable the development of strategies to maintain their vitality after cell culture. Herein, we compared the gene expression profiles of human embryonic stem cells and human BM-MSCs from donors of different ages. We first found that the expression of discoidin domain receptor 2 (DDR2) in adult donor-derived hBM-MSCs was lower than it was in the young donor-derived hBM-MSCs. Moreover, in vitro cultured late-passage hBM-MSCs showed significant downregulation of DDR2 compared to their early-passage counterparts, and siRNA inhibition of DDR2 expression recapitulated features of senescence in early-passage hBM-MSCs. Further, we found through knockdown and overexpression approaches that coactivator-associated arginine methyltransferase 1 (CARM1) regulated the expression level of DDR2 and the senescence of hBM-MSCs. Finally, chromatin immunoprecipitation analysis confirmed direct binding of CARM1 to the DDR2 promoter region with a high level of H3R17 methylation in early-passage hBM-MSCs, and inhibition of CARM1-mediated histone arginine methylation decreased DDR2 expression and led to cellular senescence. Taken together, our findings suggest that DDR2 plays a major role in regulating the in vitro senescence of hBM-MSCs and that CARM1-mediated histone H3 methylation might be the upstream regulatory mechanism controlling this function of DDR2.

The roles of DDR2 and substrate stiffness on cancer cell transcriptome and proliferation

“…Therefore, it is not surprising to observe that DDR2 is also required for proper neuroblastoma cell proliferation. Furthermore, recent work from Xu et al (48) suggested that epigenetic downregulation of DDR2 in human-derived bone marrow mesenchymal stem cells is associated with reduced proliferation and increased senescence of these cells, which is in agreement with our findings in SH-SY5Y neuroblastoma cells. Importantly, a previous work demonstrates that pharmacologic inhibitions or genetic ablation of DDR2 in several neuroblastoma cell lines impairs their proliferation and in vivo tumor growth and metastasis (Rozen et al, 2023 under revision).…”

DDR2 signaling and mechanosensing orchestrate neuroblastoma cell fate through different transcriptome mechanisms

“…This is consistent with our findings that DDR2 is also required for proper neuroblastoma cell proliferation. Furthermore, a recent study from Xu et al [57] suggested that epigenetic downregulation of DDR2 in human-derived bone marrow mesenchymal stem cells is associated with reduced proliferation and increased senescence of these cells, which is in agreement with our findings in SH-SY5Y neuroblastoma cells.…”

Molecular Mechanisms Contributing to Mesenchymal Stromal Cell Aging

“…DNA damage progressively accumulates with cell aging increasing the risk of genomic instability [2]. Accordingly, in vitro aged BM-MSCs display increased γH2AX [136]. In general, MSC phenotype is stable in culture [17,26,137]; conventional surface markers seem not modified during in vitro aging, whereas Stro-1 (stromal cell surface marker-1), CD106 (VCAM-1) and CD146 (MCAM) appear downregulated during prolonged culture [2,29].…”

“…The discoidin domain receptor 2 (DDR2, a tyrosine kinase collagen receptor expressed by MSCs) was found to be correlated with in vitro BM-MSC senescence: it plays a relevant role in BM-MSC commitment towards osteogenic differentiation rather than adipogenic; it progressively decreases with culture passages; its inhibition in low-passage BM-MSC recapitulates senescence features. DDR2 plays therefore a major role in regulating the in vitro senescence of human BM-MSCs [136]. Moreover, coactivator-associated arginine methyltransferase1 (CARM1) was found to be the upstream up-regulator of DDR2 expression via increased H3 methylation, thus contributing to rejuvenation of late passage BM-MSCs [136].…”

“…DDR2 plays therefore a major role in regulating the in vitro senescence of human BM-MSCs [136]. Moreover, coactivator-associated arginine methyltransferase1 (CARM1) was found to be the upstream up-regulator of DDR2 expression via increased H3 methylation, thus contributing to rejuvenation of late passage BM-MSCs [136]. Decreased expression of HDACs was also observed in senescent MSCs, and treatment with HDAC inhibitors has been shown to induce cellular senescence [144].…”