Basic Study
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World J Stem Cells. Apr 26, 2025; 17(4): 103482
Published online Apr 26, 2025. doi: 10.4252/wjsc.v17.i4.103482
EZH2, via an association with KDM2B, modulates osteogenic differentiation of root apical papillary stem cells
Hui-Yue Xu, Yan-Tong Wang, Hao-Qing Yang, Yang-Yang Cao, Zhi-Peng Fan
Hui-Yue Xu, Yan-Tong Wang, Hao-Qing Yang, Zhi-Peng Fan, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University, Beijing 100050, China
Yang-Yang Cao, School of Stomatology, Capital Medical University, Beijing 100050, China
Co-corresponding authors: Yang-Yang Cao and Zhi-Peng Fan.
Author contributions: Xu HY contributed to the methodology, formal analysis, and writing of the original draft of this manuscript; Xu HY and Yang HQ contributed to the software; Xu HY and Cao YY were involved in the investigation of this manuscript; Wang YT contributed to the data curation; Wang YT and Yang HQ participated in the validation; Cao YY and Fan ZP contributed to the conceptualization and supervision of this manuscript; Cao YY participated in the visualization; Fan ZP contributed to the funding acquisition, resources, and manuscript review & editing. Cao YY and Fan ZP contributed equally to this research work. Their combined expertise and collaborative efforts were critical in conceptualizing the study, designing the methodology, analyzing the data, and interpreting the results. This equal contribution reflects a balanced sharing of intellectual input, technical skills, and overall responsibility for the integrity and scientific rigor of the study. For clarity and efficiency during the manuscript submission, peer review, and publication process, Fan ZP is designated as the primary corresponding author. As the primary contact, Fan ZP will handle all communications with the journal and ensure that all procedural and editorial queries are addressed promptly. This appointment does not diminish the equal contribution of the two corresponding authors; rather, it serves to streamline administrative processes and enhance coordination throughout the publication workflow. Both authors remain jointly responsible for the accuracy, reliability, and overall quality of the research, adhering to the highest standards of scientific and academic integrity.
Supported by National Key Research and Development Program, No. 2022YFA1104401; Beijing Natural Science Foundation, No. 7222075; CAMS Innovation Fund for Medical Sciences, No. 2019RU020; and Innovation Research Team Project of Beijing Stomatological Hospital, No. CXTD202204.
Institutional review board statement: The human experiments involved in this study was approved by the Ethical Committee of Beijing Stomatological Hospital, Capital Medical University (Ethical Review No. CMUSH-IRB-KJ-PJ-2022-24).
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Ethical Committee of Beijing Stomatological Hospital, Capital Medical University (Ethical Review No. KQYY-202110-003).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Data sharing statement: All data can be provided as needed. RNA-seq and ChIP-seq data can be obtained by contacting the corresponding author.
Open Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Zhi-Peng Fan, PhD, Professor, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University, No. 4 Tiantanxili, Dongcheng District, Beijing 100050, China. zpfan@ccmu.edu.cn
Received: November 21, 2024
Revised: February 23, 2025
Accepted: April 7, 2025
Published online: April 26, 2025
Processing time: 153 Days and 19.4 Hours
Abstract
BACKGROUND

Stem cells from apical papilla (SCAPs) represent promising candidates for bone regenerative therapies due to their osteogenic potential. However, enhancing their differentiation capacity remains a critical challenge. Enhancer of zeste homolog 2 (EZH2), a histone H3 lysine 27 methyltransferase, regulates osteogenesis through epigenetic mechanisms, but its role in SCAPs remains unclear. We hypothesized that EZH2 modulates SCAP osteogenic differentiation via interaction with lysine demethylase 2B (KDM2B), offering a target for therapeutic intervention.

AIM

To investigate the functional role and molecular mechanism of EZH2 in SCAP osteogenic differentiation.

METHODS

SCAPs were isolated from healthy human third molars (n = 6 donors). Osteogenic differentiation was assessed via Alizarin red staining and alkaline phosphatase assays. EZH2 overexpression/knockdown models were established using lentiviral vectors. Protein interactions were analyzed by co-immunoprecipitation, transcriptomic changes via microarray (Affymetrix platform), and chromatin binding by chromatin immunoprecipitation-quantitative polymerase chain reaction. In vivo bone formation was evaluated in immunodeficient mice (n = 8/group) transplanted with SCAPs-hydroxyapatite scaffolds. Data were analyzed using Student’s t-test and ANOVA.

RESULTS

EZH2 overexpression increased osteogenic markers and mineralized nodule formation. In vivo, EZH2-overexpressing SCAPs generated 10% more bone/dentin-like tissue. Co-immunoprecipitation confirmed EZH2-KDM2B interaction, and peptide-mediated disruption of this binding enhanced osteogenesis. Transcriptome analysis identified 1648 differentially expressed genes (971 upregulated; 677 downregulated), with pathway enrichment in Wnt/β-catenin signaling.

CONCLUSION

EZH2 promotes SCAP osteogenesis via antagonistic interaction with KDM2B, and targeted disruption of this axis offers a translatable strategy for bone regeneration.

Keywords: Bioactive peptides; Bone tissue engineering; Enhancer of zeste homolog 2; Osteogenic; Apical papillary stem cell

Core Tip: Enhancing the osteogenic potential of stem cells is crucial for bone regeneration. This study reveals that overexpressing enhancer of zeste homolog 2 (EZH2), a key methyltransferase, promotes osteogenic differentiation in root apical papillary stem cells. EZH2 enhances osteogenesis by regulating related protein and gene expression, both in vitro and in vivo. Furthermore, the interaction between EZH2 and lysine demethylase 2B inhibits this process, suggesting that disrupting this interaction with small-molecule peptides could improve apical papillary stem cell-based therapies for bone defects. This finding opens new avenues for optimizing stem cell-based treatments through targeted molecular modulation.