Double overexpression of miR-19a and miR-20a in induced pluripotent stem cell-derived mesenchymal stem cells effectively preserves the left ventricular function in dilated cardiomyopathic rat

Jiunn-Jye Sheu; Han-Tan Chai; Pei-Hsun Sung; John Y Chiang; Tien-Hung Huang; Pei-Lin Shao; Shun-Cheng Wu; Hon-Kan Yip

doi:10.1186/s13287-021-02440-4

Double overexpression of miR-19a and miR-20a in induced pluripotent stem cell-derived mesenchymal stem cells effectively preserves the left ventricular function in dilated cardiomyopathic rat

Stem Cell Res Ther. 2021 Jun 29;12(1):371. doi: 10.1186/s13287-021-02440-4.

Authors

Jiunn-Jye Sheu^#¹, Han-Tan Chai^#², Pei-Hsun Sung^{2

3

4}, John Y Chiang^{5

6}, Tien-Hung Huang^{2

4}, Pei-Lin Shao⁷, Shun-Cheng Wu^{8

9

10}, Hon-Kan Yip^{11

12

13

14

15

16}

Affiliations

¹ Division of Thoracic and Cardiovascular Surgery, Department of Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, 83301, Taiwan.
² Division of Cardiology, Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University, College of Medicine, 123, Dapi Road, Niaosung Dist, Kaohsiung, 83301, Taiwan.
³ Center for Shockwave Medicine and Tissue Engineering, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan.
⁴ Institute for Translational Research in Biomedicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan.
⁵ Department of Computer Science and Engineering, National Sun Yat-Sen University, Kaohsiung, Taiwan.
⁶ Department of Healthcare Administration and Medical Informatics, Kaohsiung Medical University, Kaohsiung, Taiwan.
⁷ Department of Nursing, Asia University, Taichung, Taiwan.
⁸ Regenerative Medicine and Cell Therapy Research Center, Kaohsiung Medical University, No. 100, Shih-Chuan 1st Road, Kaohsiung, 807, Taiwan. shunchengwu@hotmail.com.
⁹ Orthopaedic Research Center, Kaohsiung Medical University, Kaohsiung, Taiwan. shunchengwu@hotmail.com.
¹⁰ Post-Baccalaureate Program in Nursing, Asia University, Taichung, Taiwan. shunchengwu@hotmail.com.
¹¹ Division of Cardiology, Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University, College of Medicine, 123, Dapi Road, Niaosung Dist, Kaohsiung, 83301, Taiwan. han.gung@msa.hinet.net.
¹² Center for Shockwave Medicine and Tissue Engineering, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. han.gung@msa.hinet.net.
¹³ Institute for Translational Research in Biomedicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan. han.gung@msa.hinet.net.
¹⁴ Department of Nursing, Asia University, Taichung, Taiwan. han.gung@msa.hinet.net.
¹⁵ Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, Taiwan. han.gung@msa.hinet.net.
¹⁶ Division of Cardiology, Department of Internal Medicine, Xiamen Chang Gung Hospital, Xiamen, Fujian, China. han.gung@msa.hinet.net.

^# Contributed equally.

Abstract

Background: This study tested the hypothesis that double overexpression of miR-19a and miR-20a (dOex-mIRs) in human induced pluripotent stem cell (iPS)-derived mesenchymal stem cells (MSCs) effectively preserved left ventricular ejection fraction (LVEF) in dilated cardiomyopathy (DCM) (i.e., induced by doxorubicin) rat.

Methods and results: In vitro study was categorized into groups G1 (iPS-MSC), G2 (iPS-MSC^dOex-mIRs), G3 (iPS-MSC + H₂O₂/100uM), and G4 (iPS-MSC^dOex-mIRs + H₂O₂/100uM). The in vitro results showed the cell viability was significantly lower in G3 than in G1 and G2, and that was reversed in G4 but it showed no difference between G1/G2 at time points of 6 h/24 h/48 h, whereas the flow cytometry of intra-cellular/mitochondrial oxidative stress (DCFA/mitoSOX) and protein expressions of mitochondrial-damaged (cytosolic-cytochrome-C/DRP1/Cyclophilin-D), oxidative-stress (NOX-1/NOX2), apoptotic (cleaved-caspase-3/PARP), fibrotic (p-Smad3/TGF-ß), and autophagic (ratio of LC3B-II/LC3BI) biomarkers exhibited an opposite pattern of cell-proliferation rate (all p< 0.001). Adult-male SD rats (n=32) were equally divided into groups 1 (sham-operated control), 2 (DCM), 3 (DCM + iPS-MSCs/1.2 × 10⁶ cells/administered by post-28 day's DCM induction), and 4 (DCM + iPS-MSC^dOex-mIRs/1.2 × 10⁶ cells/administered by post-28 day's DCM induction) and euthanized by day 60 after DCM induction. LV myocardium protein expressions of oxidative-stress signaling (p22-phox/NOX-1/NOX-2/ASK1/p-MMK4,7/p-JNK1,2/p-cJUN), upstream (TLR-4/MAL/MyD88/TRIF/TRAM/ TFRA6/IKK_α/ß/NF-κB) and downstream (TNF-α/IL-1ß/MMP-9) inflammatory signalings, apoptotic (cleaved-PARP/mitochondrial-Bax), fibrotic (Smad3/TGF-ß), mitochondrial-damaged (cytosolic-cytochrome-C/DRP1/cyclophilin-D), and autophagic (beclin1/Atg5) biomarkers were highest in group 2, lowest in group 1 and significantly lower in group 4 than in group 3, whereas the LVEF exhibited an opposite pattern of oxidative stress (all p< 0.0001).

Conclusion: iPS-MSC^dOex-mIRs therapy was superior to iPS-MSC therapy for preserving LV function in DCM rat.

Keywords: Dilated cardiomyopathy; Double overexpression of microRNAs; Inflammation; Mitochondrial damage; Oxidative stress.

Publication types

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

MeSH terms

Animals
Cardiomyopathy, Dilated* / genetics
Cardiomyopathy, Dilated* / therapy
Humans
Hydrogen Peroxide
Induced Pluripotent Stem Cells*
Male
Mesenchymal Stem Cells*
MicroRNAs* / genetics
Rats
Rats, Sprague-Dawley
Stroke Volume
Ventricular Function, Left

Substances

MIRN19 microRNA, human
MIRN20a microRNA, human
MicroRNAs
Hydrogen Peroxide