BMSC-derived exosomal miR-27a-3p and miR-196b-5p regulate bone remodeling in ovariectomized rats

PeerJ. 2022 Sep 22;10:e13744. doi: 10.7717/peerj.13744. eCollection 2022.

Abstract

Background: In the bone marrow microenvironment of postmenopausal osteoporosis (PMOP), bone marrow mesenchymal stem cell (BMSC)-derived exosomal miRNAs play an important role in bone formation and bone resorption, although the pathogenesis has yet to be clarified.

Methods: BMSC-derived exosomes from ovariectomized rats (OVX-Exo) and sham-operated rats (Sham-Exo) were co-cultured with bone marrow-derived macrophages to study their effects on osteoclast differentiation. Next-generation sequencing was utilized to identify the differentially expressed miRNAs (DE-miRNAs) between OVX-Exo and Sham-Exo, while target genes were analyzed using bioinformatics. The regulatory effects of miR-27a-3p and miR-196b-5p on osteogenic differentiation of BMSCs and osteoclast differentiation were verified by gain-of-function and loss-of-function analyses.

Results: Osteoclast differentiation was significantly enhanced in the OVX-Exo treatment group compared to the Sham-Exo group. Twenty DE-miRNAs were identified between OVX-Exo and Sham-Exo, among which miR-27a-3p and miR-196b-5p promoted the expressions of osteogenic differentiation markers in BMSCs. In contrast, knockdown of miR-27a-3p and miR-196b-5p increased the expressions of osteoclastic markers in osteoclast. These 20 DE-miRNAs were found to target 11435 mRNAs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that these target genes were involved in several biological processes and osteoporosis-related signaling pathways.

Conclusion: BMSC-derived exosomal miR-27a-3p and miR-196b-5p may play a positive regulatory role in bone remodeling.

Keywords: Bone marrow mesenchymal stem cell; Exosomes; MicroRNAs; Next-generation sequencing; Osteoclast; Osteogenic differentiation; Postmenopausal osteoporosis.

Grant support

This work was supported by Shenzhen Fundamental Research Key Project (No. JCYJ20200109150641992), Shenzhen Science and Technology Innovation Committee (No. JCYJ20170818164059405), the Key-Area Research and Development Program of Guangdong Province, China (No. 2020B010165004) and Category A of Shenzhen Hong Kong Jointly Funded Project (No. SGDX20201103095600002). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.