Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Apr 26, 2023; 15(4): 248-267
Published online Apr 26, 2023. doi: 10.4252/wjsc.v15.i4.248
Bone marrow mesenchymal stem cell-derived exosomal microRNAs target PI3K/Akt signaling pathway to promote the activation of fibroblasts
Fang-Qi Li, Wen-Bo Chen, Zhi-Wen Luo, Yi-Sheng Chen, Ya-Ying Sun, Xiao-Ping Su, Jun-Ming Sun, Shi-Yi Chen
Fang-Qi Li, Wen-Bo Chen, Zhi-Wen Luo, Yi-Sheng Chen, Ya-Ying Sun, Shi-Yi Chen, Department of Sports Medicine, Huashan Hospital, Fudan University, Shanghai 200040, China
Xiao-Ping Su, Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
Jun-Ming Sun, Laboratory Animal Center, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
Author contributions: Li FQ, Chen WB, and Chen YS designed and performed most of the experiments, analyzed and interpreted the data, and wrote the manuscript; Luo ZW and Sun YY assisted during the acquisition, analysis, and interpretation of data and revised the manuscript; and all authors have reviewed and approved the final manuscript. Li FQ, Chen WB, and Luo ZW contributed equally to this study. Su XP (xiaopingsu2013@aliyun.com), Sun JM (sjm990205@163.com), and Chen SY (cshiyi@163.com) correspond to this study.
Supported by Sanming Project of Medicine in Shenzhen, No. SZSM201612078; Health Shanghai Initiative Special Fund (Medical-Sports Integration, Creating a New Model of Exercise for Health), No. JKSHZX-2022-02.
Institutional review board statement: This study involves no human or animal subjects.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
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: Shi-Yi Chen, Doctor, MD, PhD, Academic Editor, Academic Fellow, Chairman, Department of Sports Medicine, Huashan Hospital, Fudan University, No. 12 Middle Wulumuqi Road, Jing’an District, Shanghai 200040, China. cshiyi@163.com
Received: November 23, 2022
Peer-review started: November 23, 2022
First decision: January 6, 2023
Revised: January 19, 2023
Accepted: March 23, 2023
Article in press: March 23, 2023
Published online: April 26, 2023
Processing time: 153 Days and 15.3 Hours
ARTICLE HIGHLIGHTS
Research background

The normal tendon-bone insertion has a transitional structure consisting of four gradated layers including bone, mineralized fibrocartilage layer, non-mineralized fibrocartilage layer and tendon. This complex structure can disperse force from bone to tendon, preventing stress concentration. However, current therapeutic strategies cannot restore this structure, hence raising the risk of re-injury.

Research motivation

Recent evidence indicates that conditioned medium, primarily contains exosomes of mesenchymal stem cells (MSCs), can stimulate the activation of fibroblasts, thereby promoting tendon-bone healing. However, the underlying mechanism is not comprehensively understood. Moreover, the expression of exosome microRNA (miRNA) may vary depending on the status of MSCs, which presents difficulties in identifying the roles associated with MSC-derived exosomal miRNAs.

Research objectives

To identify overlapped bone marrow MSC (BMSC)-derived exosomal miRNAs in three GSE datasets and verify their effects as well as mechanisms on fibroblasts.

Research methods

BMSC-derived exosomal miRNAs data were downloaded from the Gene Expression Omnibus database. The candidate miRNAs were obtained by the intersection of different datasets. TargetScan was used to predict potential target genes. Functional and pathway analyses were conducted using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases. Hub genes in the protein-protein interaction (PPI) network were analyzed using Cytoscape software. The transcripts of the target gene were assessed by dual luciferase reporter assays. BrdU, wound healing assay, collagen contraction assay and the expression of COL I and α-smooth muscle actin positive were applied to investigate the cell proliferation, migration and collagen synthesis.

Research results

Bioinformatics analyses showed two BMSC-derived exosomal miRNAs, has-miR-144-3p and has-miR-23b-3p, were overlapped in three GSE datasets. PPI network analysis and functional enrichment analyses indicated that both miRNAs regulated the PI3K/Akt signaling pathway by targeting phosphatase and tensin homolog (PTEN). In vitro experiments confirmed that both miRNAs stimulated proliferation, migration and collagen synthesis of NIH3T3 fibroblasts. Interfering with PTEN affected the phosphorylation of Akt and thus activated fibroblasts.

Research conclusions

BMSC-derived exosomes promote fibroblast activation possibly through the PTEN and PI3K/Akt signaling pathways, which may serve as potential targets to further promote tendon-bone healing.

Research perspectives

Further exploration is needed to pinpoint the specific functions of exosomal miRNAs. In vivo studies may better reveal the effect of exosomal miRNAs on tendon-bone healing.