Bone marrow mesenchymal stem cells (BMSCs)-derived exosomal METTL3 regulates the m6A methylation of SMAD5 to promote osteogenic differentiation of osteoblasts.

IF 2.8 4区医学 Q3 CELL BIOLOGY

Connective Tissue Research Pub Date : 2025-04-29 DOI:10.1080/03008207.2025.2496832

Zhenhua Li, Yifei Liu, Xiulan Zhao, Guohua Xu

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引用次数: 0

Abstract

Background: Methyltransferase-like 3 (METTL3) is implicated in human diseases, including osteoporosis (OP). In this study, we aimed to explore the functions and mechanisms of METTL3 in OP using bone marrow mesenchymal stem cells (BMSCs).

Methods: The identification of BMSCs-derived exosomes was conducted by transmission electron microscope (TEM), Nanoparticle Tracking Analysis (NTA) and western blot. The osteogenic differentiation of osteoblasts (hFOB1.19) was analyzed by Alizarin red staining assay, Alkaline phosphatase (ALP) staining assay and western blot. The relationship between METTL3 and SMAD family member 5 (SMAD5) was analyzed by Methylated RNA Immunoprecipitation (MeRIP) assay and dual-luciferase reporter assay.

Results: BMSCs-derived exosomes (BMSC-Exos) promoted the osteogenic differentiation and elevated METTL3 expression in hFOB1.19 cells. Exosomal METTL3 knockdown repressed the osteogenic differentiation in hFOB1.19 cells. METTL3 could stabilize and regulate SMAD5 expression by N6-methyladenosine (m6A) modification. Moreover, SMAD5 overexpression restored exosomal METTL3 knockdown-mediated effect on the osteogenic differentiation in hFOB1.19 cells.

Conclusion: BMSCs-derived exosomal METTL3 mediated the m6A methylation of SMAD5 to facilitate osteogenic differentiation of hFOB1.19 cells.

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骨髓间充质干细胞（BMSCs）来源的外泌体METTL3调节SMAD5的m6A甲基化，促进成骨细胞的成骨分化。

背景：甲基转移酶样3 （METTL3）与人类疾病有关，包括骨质疏松症（OP）。在这项研究中，我们旨在探讨METTL3在骨髓间充质干细胞（BMSCs）的OP中的功能和机制。方法：采用透射电镜（TEM）、纳米颗粒跟踪分析（NTA）和western blot方法对bmscs来源的外泌体进行鉴定。采用茜素红染色法、碱性磷酸酶（ALP）染色法和western blot分析成骨细胞（hFOB1.19）的成骨分化情况。采用甲基化RNA免疫沉淀法（MeRIP）和双荧光素酶报告基因法分析METTL3与SMAD家族成员5 （SMAD5）的关系。结果：bmscs来源的外泌体（bmscs - exos）促进hFOB1.19细胞成骨分化，升高METTL3表达。外泌体METTL3敲低抑制hFOB1.19细胞的成骨分化。METTL3可以通过n6 -甲基腺苷（m6A）修饰稳定和调节SMAD5的表达。此外，SMAD5过表达恢复了外泌体METTL3敲低介导的对hFOB1.19细胞成骨分化的影响。结论：bmscs来源的外泌体METTL3介导SMAD5的m6A甲基化，促进hFOB1.19细胞的成骨分化。

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来源期刊

Connective Tissue Research 生物-细胞生物学

CiteScore

6.60

自引率

3.40%

发文量

审稿时长

2 months

期刊介绍： The aim of Connective Tissue Research is to present original and significant research in all basic areas of connective tissue and matrix biology. The journal also provides topical reviews and, on occasion, the proceedings of conferences in areas of special interest at which original work is presented. The journal supports an interdisciplinary approach; we present a variety of perspectives from different disciplines, including Biochemistry Cell and Molecular Biology Immunology Structural Biology Biophysics Biomechanics Regenerative Medicine The interests of the Editorial Board are to understand, mechanistically, the structure-function relationships in connective tissue extracellular matrix, and its associated cells, through interpretation of sophisticated experimentation using state-of-the-art technologies that include molecular genetics, imaging, immunology, biomechanics and tissue engineering.