Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Aug 26, 2023; 15(8): 821-841
Published online Aug 26, 2023. doi: 10.4252/wjsc.v15.i8.821
Wnt signaling pathway inhibitor promotes mesenchymal stem cells differentiation into cardiac progenitor cells in vitro and improves cardiomyopathy in vivo
Rabbia Muneer, Rida-e-Maria Qazi, Abiha Fatima, Waqas Ahmad, Asmat Salim, Luciana Dini, Irfan Khan
Rabbia Muneer, Rida-e-Maria Qazi, Abiha Fatima, Waqas Ahmad, Asmat Salim, Irfan Khan, Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Sindh, Pakistan
Luciana Dini, Department of Biology and Biotechnologies Charles Darwin, Sapienza University of Rome, Rome 00185, Italy
Author contributions: Muneer R performed experiments and wrote the original manuscript; Qazi R, Fatima A, and Ahmad W helped in experimentation and writing; Salim A evaluated and analyzed the data, and reviewed the manuscript; Dini L evaluated and analyzed the data; Khan I conceived and designed the studies, evaluated and analyzed the data, and finalized the manuscript.
Institutional review board statement: The independent ethical committee of PCMD, ICCBS, University of Karachi, for human subjects has approved the protocol, No. ICCBS/IEC-067-HT/UCB-2021/Protocol/1.0.
Institutional animal care and use committee statement: Ethical approval for the use of animals was obtained from the Institutional Animal Care and Use Committee at the PCMD, ICCBS, University of Karachi with protocol number 2021-006.
Informed consent statement: Informed consent was obtained from the donor.
Conflict-of-interest statement: Authors declare no conflict of interest.
Data sharing statement: No additional data to share.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Irfan Khan, PhD, Assistant Professor, Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, University Road, Karachi 75270, Sindh, Pakistan. khan@iccs.edu
Received: March 26, 2023
Peer-review started: March 26, 2023
First decision: May 17, 2023
Revised: May 31, 2023
Accepted: July 3, 2023
Article in press: July 3, 2023
Published online: August 26, 2023
Processing time: 151 Days and 23.6 Hours
ARTICLE HIGHLIGHTS
Research background

Cardiovascular diseases particularly myocardial infarction (MI) is a global health complication with high mortality and morbidity rate. As cardiac tissue lacks regeneration potential, so cardiac tissue regeneration using a potent small molecule inhibitor Wnt production-4 (IWP-4) for stem cell fate transition towards cardiomyocytes could be an effective approach.

Research motivation

Inhibition of Wnt pathway is important in stem cell fate determination towards cardiomyocytes. Wnt pathway inhibitor, such as IWP-4, may promote the differentiation of mesenchymal stem cells (MSCs) into cardiac lineage. These pre-conditioned cells may provide better survival, homing and migration capability at the site of injury.

Research objectives

This study was designed to evaluate the IWP-4 cardiac differentiation capability and its subsequent in vivo effects.

Research methods

Human umbilical cord-derived MSCs were characterized on the basis of morphology, immunophenotyping of surface markers associated with MSCs and tri-lineage differentiation capability. Isolated MSCs were treated with 5 μM IWP-4 at two different time intervals. Cardiomyogenic differentiation of treated MSCs was evaluated at DNA and protein levels. MI rat model was developed. IWP-4 treated as well as untreated MSCs were implanted in the MI model, and cardiac function was analyzed via echocardiography. MSCs were labeled with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) dye for tracking, and regeneration of the infarcted myocardium was examined by histology and immunohistochemistry.

Research results

Isolated MSCs were characterized and then pre-conditioned with 5 μM concentration of IWP-4. The cardiac specific gene and protein expression analysis exhibited more remarkable results in fourteen days treated group that was eventually selected for in vivo transplantation. Cardiac function was restored in the IWP-4 treated group in comparison to the MI group. Immunohistochemical analysis confirmed the homing of pre-differentiated MSCs that were labeled with DiI cell labeling dye. Histological analysis confirmed the significant reduction in the fibrotic area, and improved the left ventricular wall thickness in the IWP-4 treated MSC group.

Research conclusions

Our data suggest that treatment of MSCs with IWP-4 inhibits Wnt pathway and promotes cardiac differentiation. These pre-conditioned MSCs transplanted in vivo improved cardiac function by cell homing, survival, and differentiation at the infarcted region, increased left ventricular wall thickness, and reduced infarct size.

Research perspectives

The study demonstrated that treatment with IWP-4 improves the efficacy of the MSCs to ameliorate the cardiac function of the MI rats, as evidenced by an increase in the ejection fraction. Pre-conditioning of MSCs with IWP-4 may serve as a promising strategy to treat heart disease. Targeting the Wnt signaling pathway may represent a promising therapeutic approach for this debilitating condition.