Non-coding RNA-activated by DNA damage promotes proliferation and inhibits apoptosis of induced pluripotent stem cell-derived cardiomyocytes

doi:10.12307/2026.192

Abstract

Abstract: BACKGROUND: Although cell transplantation offers a promising approach for the treatment of myocardial infarction, the low transplantation rate limits its application. Therefore, promoting the proliferation of transplanted cells and reducing apoptosis are the key issues to be solved urgently to improve the therapeutic effect.
OBJECTIVE: To investigate the effects of non-coding RNA-activated by DNA damage (NORAD) on the proliferation of human induced pluripotent stem cell-derived cardiomyocytes and their apoptosis induced by oxygen-glucose deprivation/reoxygenation, as well as the effects of transplanting NORAD-overexpressing human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-NORADOECMs) on cardiac function in a murine model of myocardial infarction.
METHODS: The expression of NORAD in the hearts of mice at different ages (3 days old and 8 weeks old) was measured by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). A cellular model of hiPSC-NORADOECMs was established by infecting human induced pluripotent stem cells with a lentiviral vector designed to specifically upregulate NORAD, followed by directed differentiation. These cells were divided into a control group (infected with a control lentivirus) and an experimental group (infected with a NORAD-overexpressing lentivirus). The efficiency of NORAD overexpression was detected by RT-qPCR. The expression level of proliferation marker protein Ki67 was detected by cellular immunofluorescence. Intracellular reactive oxygen species levels were measured by flow cytometry after cell apoptosis was induced by oxygen-glucose deprivation/reoxygenation induction. The expression levels of Bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), and cleaved caspase-3 were analyzed by western blot assay. hiPSC-NORADOECMs were transplanted into the infarcted area of the myocardial infarction mouse model. The cardiac function was evaluated by echocardiography four weeks later.
RESULTS AND CONCLUSION: (1) NORAD expression was significantly higher in the hearts of 3-day-old neonatal mice compared with 8-week-old adult mice. (2) The differentiation of human induced pluripotent stem cells into spontaneously contracting human induced pluripotent stem cell-derived cardiomyocytes was successfully achieved. (3) A stable human induced pluripotent stem cell-derived cardiomyocytes model overexpressing NORAD (hiPSC-NORADOECMs) was successfully established. (4) The expression of the proliferation marker Ki67 was significantly increased in the NORAD-overexpressing group compared with controls. (5) Compared with controls, NORAD overexpression reduced reactive oxygen species accumulation induced by oxygen-glucose deprivation/reoxygenation in human induced pluripotent stem cell-derived cardiomyocytes; Bax and cleaved caspase-3 levels were decreased, while Bcl-2 expression was upregulated. (6) Transplantation of hiPSC-NORADOECMs significantly enhanced cardiac function in a murine model of myocardial infarction. Collectively, these findings suggest that overexpression of the NORAD promotes the proliferation of human induced pluripotent stem cell-derived cardiomyocytes, reduces reactive oxygen species accumulation, and inhibits apoptosis following oxygen-glucose deprivation/reoxygenation induction, thereby enhancing the cardiac repair capacity of human induced pluripotent stem cell-derived cardiomyocytes in infarcted hearts.

Key words: long non-coding RNA, induced pluripotent stem cell, cardiomyocyte, oxygen-glucose deprivation/reoxygenation, cell proliferation, myocardial infarction, apoptosis, cell transplantation

CLC Number:

Huan Kanghui, Jiang Yujian, Bian Weihua. Non-coding RNA-activated by DNA damage promotes proliferation and inhibits apoptosis of induced pluripotent stem cell-derived cardiomyocytes[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(19): 4926-4933.

Figures/Tables 7

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