Inhibition of circALPK2 enhances proliferation and therapeutic potential of human pluripotent stem cell-derived cardiomyocytes in myocardial infarction

被引：0

作者：

Wu, Hongchun ^{[1
,2
]}

Jiang, Xue ^{[1
,2
]}

Fan, Hao ^{[1
,2
]}

Li, Jingjing ^{[1
,2
]}

Li, Yuan ^{[1
,2
]}

Lin, Yingjiong ^{[1
,2
]}

Zhao, Dandan ^{[1
,2
]}

Han, Xinglong ^{[1
,2
]}

Yu, Miao ^{[1
,2
]}

Tang, Jun-Ming ^{[3
]}

Hu, Shijun ^{[1
,2
]}

Lei, Wei ^{[1
,2
]}

机构：

[1] Soochow Univ, Affiliated Hosp 1, Dept Cardiovasc Surg, Suzhou 215000, Jiangsu, Peoples R China

[2] Soochow Univ, Collaborat Innovat Ctr Hematol, Suzhou Med Coll, State Key Lab Radiat Med & Protect,Inst Cardiovasc, Suzhou 215000, Jiangsu, Peoples R China

[3] Hubei Univ Med, Sch Basic Med Sci, Hubei Key Lab Embryon Stem Cell Res, Shiyan 442000, Hubei, Peoples R China

来源：

STEM CELL RESEARCH & THERAPY | 2025年 / 16卷 / 01期

基金：

国家重点研发计划; 中国国家自然科学基金;

关键词：

Cardiomyocyte proliferation; circALPK2; miR-9; GSK3B; Myocardial infarction; ZEBRAFISH;

D O I：

10.1186/s13287-025-04230-8

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

Background Understanding the mechanisms regulating human cardiomyocyte proliferation holds significant promise for developing effective therapies to enhance cardiac regeneration and repair. This study investigates the role of circALPK2, a circular RNA derived from the back-splicing of the 4th exon of alpha protein kinase 2 (ALPK2), in regulating cardiomyocyte proliferation and its therapeutic efficacy in myocardial infarction (MI) treatment. Methods Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) were used to assess the expression and function ofcircALPK2. Lentiviral shRNA-mediated knockdown of circALPK2 was performed in hESC-CMs, followed by RNA sequencing to identify targeted genes and biological processes. The proliferative capacity of wild-type and circALPK2 knockdown hESC-CMs was evaluated using CCK-8 assay, EdU staining and RT-qPCR analysis of cell cycle-related genes. Dual luciferase assays were conducted to validate the predicted miRNA targets and their downstream effects. For in vivo evaluation, MI mice were injected with either wild-type or circALPK2 knockdown hESC-CMs, and the therapeutic potential was assessed by echocardiographic and histological analyses. Results We identified circALPK2 as a negative regulator of cell proliferation in hESC-CMs. CircALPK2 was abundantly expressed in hESC-CMs. Knockdown of circALPK2 significantly enhanced cell proliferation in hESC-CMs, as demonstrated by CCK-8 assays (p < 0.001) and EdU staining (p < 0.001), and accelerated the expression of cell cycle-related genes, including CCNA2(p < 0.05) and CDK1 (p < 0.01). Furthermore, circALPK2 was found to function as a sponge to inhibit miR-9 activity, while miR-9 mimics significantly boosted the proliferative capacity of hESC-CMs. Glycogen synthase kinase 3 beta (GSK3B), a key inhibitor of WNT signaling, was identified as a direct target of miR-9, mediating the regulation of cardiomyocyte proliferation. Importantly, circALPK2 knockdown improved the myocardial repair potential of hESC-CMs when injected into infarcted mouse hearts, as indicated by improved left ventricular ejection fraction (p < 0.01) and fractional shortening (p < 0.05). Conclusions Our study identifies the circALPK2/miR-9/GSK3B axis as a novel target for promoting cardiomyocyte proliferation and enhancing cardiac regeneration.

引用

页数：13