NSUN2 facilitates tenogenic differentiation of rat tendon-derived stem cells via m5C methylation of KLF2

IF 3.4 3区环境科学与生态学 Q3 CELL & TISSUE ENGINEERING Regenerative Therapy Pub Date : 2024-06-01 DOI:10.1016/j.reth.2024.08.023

Wei Lin , Zhi Lin , Lizhi Wu , Youmao Zheng , Huifeng Xi

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Abstract

Introduction

Tendon-derived stem cells (TDSCs) play a critical role in tendon repair. N5-methylcytosine (m5C) is a key regulator of cellular processes such as differentiation. This study aimed to investigate the impact of m5C on TDSC differentiation and the underlying mechanism.

Methods

TDSCs were isolated from rats and identified, and a tendon injury rat model was generated. Tenogenic differentiation in vitro was evaluated using Sirius red staining and quantitative real-time polymerase chain reaction, while that in vivo was assessed using immunohistochemistry and hematoxylin‒eosin staining. m5C methylation was analyzed using methylated RNA immunoprecipitation, dual-luciferase reporter assay, and RNA stability assay.

Results

The results showed that m5C levels and NSUN2 expression were increased in TDSCs after tenogenic differentiation. Knockdown of NSUN2 inhibited m5C methylation of KLF2 and decreased its stability, which was recognized by YBX1. Moreover, interfering with KLF2 suppressed tenogenic differentiation of TDSCs, which could be abrogated by KLF2 overexpression. Additionally, TDSCs after NSUN2 overexpression contributed to ameliorating tendon injury in vivo. In conclusion, NSUN2 promotes tenogenic differentiation of TDSCs via m5C methylation of KLF2 and accelerates tendon repair.

Conclusions

The findings suggest that overexpression of NSUN2 can stimulate the differentiation ability of TDSCs, which can be used in the treatment of tendinopathy.

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NSUN2 通过 KLF2 的 m5C 甲基化促进大鼠肌腱衍生干细胞的成腱分化

导言肌腱衍生干细胞（TDSCs）在肌腱修复中发挥着关键作用。N5-甲基胞嘧啶（m5C）是细胞分化等过程的关键调节因子。本研究旨在探讨 m5C 对 TDSC 分化的影响及其内在机制。采用甲基化 RNA 免疫沉淀法、双荧光素酶报告分析法和 RNA 稳定性分析法对 m5C 甲基化进行分析。敲除 NSUN2 可抑制 KLF2 的 m5C 甲基化并降低其稳定性，YBX1 可识别这种甲基化。此外，干扰 KLF2 可抑制 TDSCs 的成腱分化，而过表达 KLF2 则可抑制成腱分化。此外，NSUN2过表达后的TDSCs有助于改善体内肌腱损伤。总之，NSUN2 可通过 KLF2 的 m5C 甲基化促进 TDSCs 的腱源性分化，加速肌腱修复。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Regenerative Therapy Engineering-Biomedical Engineering

CiteScore

6.00

自引率

2.30%

发文量

106

审稿时长

49 days

期刊介绍： Regenerative Therapy is the official peer-reviewed online journal of the Japanese Society for Regenerative Medicine. Regenerative Therapy is a multidisciplinary journal that publishes original articles and reviews of basic research, clinical translation, industrial development, and regulatory issues focusing on stem cell biology, tissue engineering, and regenerative medicine.