Research Paper Volume 13, Issue 19 pp 23361—23375
piRNA-36741 regulates BMP2-mediated osteoblast differentiation via METTL3 controlled m6A modification
- 1 Surgery Department, Shaanxi Provincial People’s Hospital, Xi’an 710068, China
- 2 Department of Orthopedics, Shaanxi Provincial People’s Hospital, Xi’an 710068, China
Received: June 25, 2021 Accepted: September 20, 2021 Published: October 13, 2021
https://doi.org/10.18632/aging.203630How to Cite
Copyright: © 2021 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
The osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) is essential for bone formation, and its imbalance can lead to bone diseases such as osteoporosis. It is reported that PIWI-interacting RNA-36741 (piR-36741) is up-regulated during the osteogenic differentiation, but its role in regulating osteogenic differentiation remains unclear. Here, the primary human BMSCs were used to induce osteogenic differentiation, and the expression of piR-36741 and METTL3 (methyltransferase like 3) was up-regulated during the osteogenic differentiation of BMSCs. Moreover, interference with piR-36741 or METTL3 markedly hindered the osteogenic differentiation of BMSCs, which was manifested by a reduction in osteoblast marker expression (including RUNX2, COL1A1, OPN and OCN), osteogenic phenotype and matrix mineralization. Mechanistically, the piR-36741-PIWIL4 complex directly interacted with METTL3 and prevented METTL3-mediated m6A modification of BMP2 mRNA transcripts, thereby promoting BMP2 expression. And overexpression of BMP2 reversed the inhibitory effect of piR-36741 silence on osteogenic differentiation and the Smad pathway activity. In addition, administration with piR-36741 mimic alleviated ovariectomy-induced osteoporosis in mice. In conclusion, piRNA-36741 overexpression promoted osteogenic differentiation of BMSCs and mitigated ovariectomy-induced osteoporosis through METTL3-mediated m6A methylation of BMP2 transcripts.
Abbreviations
BMSCs: bone marrow mesenchymal stem cells; METTL3: methyltransferase like 3; ALP: alkaline phosphatase; OP: Osteoporosis; piRNAs: PIWI-interacting RNAs; m6A: N6-methyladenosine; ECL: enhanced chemiluminescence; ARS: Alizarin red staining; RIP: RNA immunoprecipitation assay; co-IP: co-immunoprecipitation; H&E: haematoxylin and eosin; BMD: bone mineral density; RUNX2: runt-related transcription factor 2; COL1A1: collagen type I; OCN: osteocalcin; OPN: osteopontin; BMP: bone morphogenetic protein.