Cyclin D2 Overexpression Enhances the Efficacy of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes for Myocardial Repair in a Swine Model of Myocardial Infarction

• Type of Material: Article
• Authors: Meng Zhao, University of Alabama Birmingham; Yuji Nakada, University of Alabama Birmingham; Yuhua Wei, University of Alabama Birmingham; Weihua Bian, University of Alabama Birmingham; Yuxin Chu, University of Alabama Birmingham; Anton V Borovjagin, University of Alabama Birmingham; Min Xie, University of Alabama Birmingham; Wuqiang Zhu, Mayo Clinic; Thanh Nguyen, University of Alabama Birmingham; Yang Zhou, University of Alabama Birmingham; Vahid Serpooshan, Emory University; Gregory P Walcott, University of Alabama Birmingham; Jianyi Zhang, University of Alabama Birmingham
• Language: English
• Date: 2021-07-20
• Publisher: LIPPINCOTT WILLIAMS & WILKINS
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2021, Wolters Kluwer Health
• Final Published Version (URL): http://dx.doi.org/10.1161/CIRCULATIONAHA.120.049497
• Title of Journal or Parent Work: CIRCULATION
• Volume: 144
• Issue: 3
• Start Page: 210
• End Page: 228
• Grant/Funding Information: This study is supported in part by the US National Institutes of Health National Heart, Lung, and Blood Institute grants RO1 HL114120, HL131017, HL149137, and UO1 HL134764.
• Keywords: FUNCTIONAL CONSEQUENCES; induced pluripotent stem cells; VEGF; ANGIOGENESIS; MICRORNAS; muscle cells; Life Sciences & Biomedicine; Cardiac & Cardiovascular Systems; myocardial infarction; CARDIAC REPAIR; Peripheral Vascular Disease; THERAPY; HEART; Cardiovascular System & Cardiology; FIBROBLASTS; Science & Technology; cyclin D2

Abstract

Background: Human induced pluripotent stem cells with normal (wild-type) or upregulated (overexpressed) levels of CCND2 (cyclin D2) expression were differentiated into cardiomyocytes (CCND2WTCMs or CCND2OECMs, respectively) and injected into infarcted pig hearts. Methods: Acute myocardial infarction was induced by a 60-minute occlusion of the left anterior descending coronary artery. Immediately after reperfusion, CCND2WTCMs or CCND2OECMs (3×107cells each) or an equivalent volume of the delivery vehicle was injected around the infarct border zone area. Results: The number of the engrafted CCND2OECMs exceeded that of the engrafted CCND2WTCMs from 6- to 8-fold, rising from 1 week to 4 weeks after implantation. In contrast to the treatment with the CCND2WTCMs or the delivery vehicle, the administration of CCND2OECM was associated with significantly improved left ventricular function, as revealed by magnetic resonance imaging. This correlated with reduction of infarct size, fibrosis, ventricular hypertrophy, and cardiomyocyte apoptosis, and increase of vascular density and arterial density, as per histologic analysis of the treated hearts. Expression of cell proliferation markers (eg, Ki67, phosphorylated histone 3, and Aurora B kinase) was also significantly upregulated in the recipient cardiomyocytes from the CCND2OECM-treated than from the CCND2WTCM-treated pigs. The cell proliferation rate and the hypoxia tolerance measured in cultured human induced pluripotent stem cell cardiomyocytes were significantly greater after treatment with exosomes isolated from the CCND2OECMs (CCND2OEExos) than from the CCND2WTCMs (CCND2WTExos). As demonstrated by our study, CCND2OEExos can also promote the proliferation activity of postnatal rat and adult mouse cardiomyocytes. A bulk miRNA sequencing analysis of CCND2OEExos versus CCND2WTExos identified 206 and 91 miRNAs that were significantly upregulated and downregulated, respectively. Gene ontology enrichment analysis identified significant differences in the expression profiles of miRNAs from various functional categories and pathways, including miRNAs implicated in cell-cycle checkpoints (G2/M and G1/S transitions), or the mechanism of cytokinesis. Conclusions: We demonstrated that enhanced potency of CCND2OECMs promoted myocyte proliferation in both grafts and recipient tissue in a large mammal acute myocardial infarction model. These results suggest that CCND2OECMs transplantation may be a potential therapeutic strategy for the repair of infarcted hearts.