KDM4D 通过结合 RPS5 增强 SCAP 的成骨/牙骨质分化和迁移。

KDM4D enhances osteo/dentinogenic differentiation and migration of SCAPs via binding to RPS5.

机构信息

Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology, Beijing, China.

Department of Endodontics, Capital Medical University School of Stomatology, Beijing, China.

出版信息

Oral Dis. 2023 Oct;29(7):2827-2836. doi: 10.1111/odi.14479. Epub 2023 Jan 20.

DOI:10.1111/odi.14479

PMID:36579641

Abstract

OBJECTIVES

Stem cells of the apical papilla (SCAPs) provide promising candidates for dental pulp regeneration. Despite great advances in the transcriptional controls of the SCAPs fate, little is known about the regulation of SCAP differentiation.

MATERIALS AND METHODS

Short hairpin RNAs and full-length RNA were used to deplete or overexpress lysine demethylase 4D (KDM4D) gene expression. Western blotting, real-time RT-PCR, alizarin red staining, and scratch migration assays were used to study the role of KDM4D and the ribosomal protein encoded by RPS5 in SCAPs. RNA microarray, chromatin Immunoprecipitation (ChIP), and co-immunoprecipitation (Co-IP) assays were performed to explore the underlying molecular mechanisms.

RESULTS

KDM4D enhanced the osteo/dentinogenic differentiation, migration, and chemotaxis of SCAPs. The microarray results revealed that 88 mRNAs were differentially expressed in KDM4D-overexpressed SCAPs. ChIP results showed knock-down of KDM4D increased the level of H3K9me2 and H3K9me3 in CNR1 promoter region. There were 37 possible binding partners of KDM4D. KDM4D was found to combine with RPS5, which also promoted the osteo/dentinogenic differentiation, migration, and chemotaxis of SCAPs.

CONCLUSIONS

KDM4D promoted the osteo/dentinogenic differentiation and migration potential of SCAPs in combination with RPS5, which provides a therapeutic clue for improving SCAPs-based dental tissue regeneration.

摘要

目的

根尖乳头干细胞（SCAP）为牙髓再生提供了有前景的候选细胞。尽管 SCAP 命运的转录调控取得了很大进展，但对 SCAP 分化的调控知之甚少。

材料和方法

使用短发夹 RNA 和全长 RNA 来耗尽或过表达赖氨酸去甲基酶 4D（KDM4D）基因表达。使用 Western blot、实时 RT-PCR、茜素红染色和划痕迁移实验来研究 KDM4D 和核糖体蛋白 RPS5 在 SCAP 中的作用。进行 RNA 微阵列、染色质免疫沉淀（ChIP）和共免疫沉淀（Co-IP）实验以探索潜在的分子机制。

结果

KDM4D 增强了 SCAP 的成骨/牙本质向分化、迁移和趋化性。微阵列结果显示，在 KDM4D 过表达的 SCAP 中，有 88 个 mRNA 表达水平差异。ChIP 结果显示，敲低 KDM4D 增加了 CNR1 启动子区域的 H3K9me2 和 H3K9me3 水平。有 37 个可能与 KDM4D 结合的伴侣蛋白。发现 KDM4D 与 RPS5 结合，这也促进了 SCAP 的成骨/牙本质向分化、迁移和趋化性。