DNA 메틸화효소 1 억제제인 RG108의 골수유래 중간엽줄기세포에서의 노화 예방효과에 대한 연구

Abstract

Mesenchymal stromal cells (MSCs) are characterized by their multipotency capacity, which allows them to differentiate to diverse cell types, and secrete a variety of trophic factors. These features indicate that MSCs might be of use in stem-cell therapy. However, MSCs undergo cellular senescence during long-term expansion, and this is accompanied by functional declines in stem-cell potency. Recent studies have shown that alteration of DNA methylation is highly associated with cellular senescence and aging-related neurodegenerative disorders, such as amyotrophic lateral sclerosis (ALS). Remedy of the altered methylation pattern may provide beneficial efficacy in these diseases. In this study, I used a DNA methyltransferase 1 inhibitor, RG108, to investigate the anti-senescence effects in human bone marrow mesenchymal stromal cells (hBM-MSCs). First, I determined the optimized dose and time of RG108 treatment in hBM-MSCs to be 5 μM for 48 h, respectively. Under these conditions, the anti-senescence genes TERT, bFGF, VEGF, and ANG were increased, whereas the senescence-related genes ATM, p21, and p53 were decreased. The number of β-galactosidase positive cells was significantly decreased in RG108-treated hBM-MSCs, whereas the rates of cell migration and cellular protection were increased. I have shown that RG108 significantly induces the expression of TERT by blocking methylation at the TERT promoter region. Thus, these data indicate that an optimized dose of RG108 may improve the cell migration, protection, and cellular senescence, which may provide a better efficacy of these cells in stem cell therapy. Next, I investigated correlation between excessive DNMT1 expression and functional decline in ALS-patient derived BM-MSCs (ALS-MSCs). The DNMT1 inhibitor RG108 was used for this study. RG108 treated ALS-MSCs exhibit increased expression of the anti-senescence genes TERT, VEGF, and ANG, and decreased expression of the senescence-related genes ATM, p21 and p53. The activity of Senescence associated β-galactosidase and the expression of senescence proteins p53 and p16 were reduced in RG108 treated ALS-MSCs. The abilities of cell migration and protection against oxidative damage were improved in the RG108 treated ALS-MSCs. In neuronal differentiation experiments, the RG108 treated ALS-MSCs more effectively differentiated into neuron-like cells. These results suggest that ALS-MSCs function can be restored by inhibiting excessively expressed DNMT1, an approach that may ultimately provide better efficacy in stem cell therapy. In the present studies suggest that DNMT1 inhibitor RG108 can improve stem cell potency and, ameliorates cellular senescence in Normal and ALS patients derived BM-MSCs. Which may provide better efficacy in stem cell therapy.