Abstract 15481: Abrogation of Senescence-associated Microrna-195 in Aged Skeletal Myoblasts Facilitates Reprogramming to Produce Induced Pluripotent Stem Cells
Background: Older age is the major risk factors for heart failure, and reprogramming a patient’s own cells to produce induced pluripotent stem cells (iPSCs) is a promising strategy for autologous cell transplantation therapy. However, low reprogramming efficiency of senescent cells remains as a major pitfall. Recently, we have shown that inhibition of senescence-associated miR-195 rejuvenated aged stem cells by reactivating telomerase reverse transcriptase (Tert). This study investigated the effects of abrogation of miR-195 expression on the reprogramming efficiency of old skeletal myoblasts (OSkMs).
Methods and Results: MiR-195 expression was significantly higher in OSkMs isolated from aged mice (24 months) as compared to SkMs from young mice (2 months), as examined by RT-PCR. In addition, OSkMs showed impaired expression of anti-aging factors (Tert and Sirt1) and higher expression of pro-aging markers (p53, p21, p16). Intriguingly, blockage of miR-195 expression in OSkMs by transfection with anti-miR-195 significantly induced expression of Tert and Sirt1 as well as telomere re-lengthening as examined by RT-PCR and quantitative fluorescent in situ hybridization (Q-FISH). It is important to note that lower reprogramming efficiency of OSkMs was improved by miR-195 abrogation. Notably, inhibition of miR-195 did not alter karyotype or expression of pluripotency markers, and iPSCs lacking miR-195 successfully differentiated into all three germ layers, indicating that deletion of miR-195 does not affect pluripotency. Furthermore, contraction rates were markedly higher in beating cells transfected with anti-miR-195 as compared to that with scramble (68.5±5.6 vs 47.3±2.8/min).
Conclusions: Abrogation of age-induced miR-195 is a novel promising approach for efficient iPSCs generation from senescent cells, which will contribute to successful autologous transplantation therapy in elderly patients.
Author Disclosures: H. Kondo: None. H. Kim: None. R.W. Millard: None. Y. Wang: None.
- © 2014 by American Heart Association, Inc.