Lack of Remuscularization Following Transplantation of Human Embryonic Stem Cell-Derived Cardiovascular Progenitor Cells in Infarcted Nonhuman Primates

Keyang Zhu; Qiang Wu; Cheng Ni; Peng Zhang; Zhiwei Zhong; Yan Wu; Yingchao Wang; Yinchuan Xu; Minjian Kong; Haifeng Cheng; Zhihua Tao; Qian Yang; He Liang; Yun Jiang; Qingju Li; Jing Zhao; Jijun Huang; Fengjiang Zhang; Qi Chen; Yi Li; Jinghai Chen; Wei Zhu; Hong Yu; Jianyi Zhang; Huang-Tian Yang; Xinyang Hu; Jian'an Wang

doi:10.1161/CIRCRESAHA.117.311578

Lack of Remuscularization Following Transplantation of Human Embryonic Stem Cell-Derived Cardiovascular Progenitor Cells in Infarcted Nonhuman Primates

Circ Res. 2018 Mar 30;122(7):958-969. doi: 10.1161/CIRCRESAHA.117.311578. Epub 2018 Jan 17.

Authors

Keyang Zhu¹, Qiang Wu¹, Cheng Ni¹, Peng Zhang¹, Zhiwei Zhong¹, Yan Wu¹, Yingchao Wang¹, Yinchuan Xu¹, Minjian Kong¹, Haifeng Cheng¹, Zhihua Tao¹, Qian Yang¹, He Liang¹, Yun Jiang¹, Qingju Li¹, Jing Zhao¹, Jijun Huang¹, Fengjiang Zhang¹, Qi Chen¹, Yi Li¹, Jinghai Chen¹, Wei Zhu¹, Hong Yu¹, Jianyi Zhang¹, Huang-Tian Yang², Xinyang Hu², Jian'an Wang²

Affiliations

¹ From the Department of Cardiology (K.Z., C.N., Z.Z., Y. Wu, Y. Wang, Y.X., Q.Y., Q.L., J. Zhao, J.C., W.Z., H.Y., X.H., J.W.), Department of Cardiovascular Surgery (M.K., H.C.), Department of Laboratory Medicine (Z.T.), and Department of Anesthesiology (F.Z., Q.C., Y.L.), Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, PR China; Key Laboratory of Stem Cell Biology and Laboratory of Molecular Cardiology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (CAS) Shanghai, University of CAS, Beijing, PR China (Q.W., P.Z., H.L., Y.J., J.H., H.-T.Y.); Cardiovascular Key Laboratory of Zhejiang Province, Hangzhou, PR China (K.Z., C.N., Z.Z., Y. Wu, Y. Wang, Y.X., Q.L., J. Zhao, J.C., W.Z., H.Y., X.H., Y.W.); and Department of Biomedical Engineering, University of Alabama at Birmingham (J. Zhang).
² From the Department of Cardiology (K.Z., C.N., Z.Z., Y. Wu, Y. Wang, Y.X., Q.Y., Q.L., J. Zhao, J.C., W.Z., H.Y., X.H., J.W.), Department of Cardiovascular Surgery (M.K., H.C.), Department of Laboratory Medicine (Z.T.), and Department of Anesthesiology (F.Z., Q.C., Y.L.), Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, PR China; Key Laboratory of Stem Cell Biology and Laboratory of Molecular Cardiology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (CAS) Shanghai, University of CAS, Beijing, PR China (Q.W., P.Z., H.L., Y.J., J.H., H.-T.Y.); Cardiovascular Key Laboratory of Zhejiang Province, Hangzhou, PR China (K.Z., C.N., Z.Z., Y. Wu, Y. Wang, Y.X., Q.L., J. Zhao, J.C., W.Z., H.Y., X.H., Y.W.); and Department of Biomedical Engineering, University of Alabama at Birmingham (J. Zhang). wangjianan111@zju.edu.cn hxy0507@zju.edu.cn htyang@sibs.ac.cn.

PMID: 29343525
DOI: 10.1161/CIRCRESAHA.117.311578

Abstract

Rationale: Human pluripotent stem cell-derived cardiovascular progenitor cells (hPSC-CVPCs) should be thoroughly investigated in large animal studies before testing in clinical trials.

Objective: The main of this study is to clarify whether hPSC-CVPCs can engraft for long time in the heart of primates after myocardial infarction (MI) and compare the effectiveness and safety of immunosuppression with cyclosporine alone or multiple-drug regimen (MDR) containing cyclosporine, methylprednisolone, and basiliximab in cynomolgus monkeys that had received intramyocardial injections of 1×10⁷ EGFP (enhanced green fluorescent protein)-expressing hPSC-CVPCs after MI. A third group of animals received the immunosuppression MDR but without cell therapy after MI (MI+MDR group).

Methods and results: Measurements of EGFP gene levels and EGFP immunofluorescence staining indicated that the hPSC-CVPC engraftment rate was greater in the MI+MDR+CVPC group than that in the MI+cyclosporine+CVPC group. However, even in the MI+MDR+CVPC group, no transplanted cells could be detected at 140 days after transplantation. Concomitantly, immunofluorescent analysis of CD3, CD4, and CD8 expression indicated that T-lymphocyte infiltration in the CVPC-transplanted hearts was less in the MDR-treated animals than in the cyclosporine-alone-treated animals. The recovery of left ventricular function on day 28 post-MI in the MI+MDR+CVPC group was better than that in the MI+MDR group. Apoptotic cardiac cells were also less common in the MI+MDR+CVPC group than in the MI+MDR group, although both immunosuppression regimens were associated with transient hepatic dysfunction.

Conclusions: This is the largest study of hPSCs in nonhuman primates in cardiovascular field to date (n=32). Compared with cyclosporine alone, MDR attenuates immune rejection and improves survival of hPSC-CVPCs in primates; this is associated with less apoptosis of native cardiac cells and better recovery of left ventricular function at 28 days. However, even with MDR, transplanted hPSC-CVPCs do not engraft and do not survive at 140 days after transplantation, thereby excluding remuscularization as a mechanism for the functional effect.

Keywords: human embryonic stem cells; immunosuppression; primates; transplantation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
Cyclosporine / administration & dosage
Cyclosporine / adverse effects
Human Embryonic Stem Cells / cytology*
Humans
Immunosuppression Therapy / adverse effects
Immunosuppression Therapy / methods
Immunosuppressive Agents / administration & dosage
Immunosuppressive Agents / adverse effects
Macaca fascicularis
Male
Muscle Development*
Myoblasts, Cardiac / cytology
Myoblasts, Cardiac / transplantation*
Myocardial Infarction / therapy*
Stem Cell Transplantation / adverse effects
Stem Cell Transplantation / methods*

Substances

Immunosuppressive Agents
Cyclosporine