PMID- 38042362 OWN - NLM STAT- MEDLINE DCOM- 20240301 LR - 20240301 IS - 1778-7254 (Electronic) IS - 1297-319X (Linking) VI - 91 IP - 2 DP - 2024 Mar TI - Circular RNA circ-3626 promotes bone formation by modulating the miR-338-3p/Runx2 axis. PG - 105669 LID - S1297-319X(23)00148-3 [pii] LID - 10.1016/j.jbspin.2023.105669 [doi] AB - OBJECTIVE: Disorders of bone homeostasis are the key factors leading to metabolic bone disease, such as senile osteoporosis, which is characterized by age-related bone loss. Bone marrow stromal cells (BMSCs) possess high osteogenic capacity which has been regarded as a practical approach to preventing bone loss. Previous studies have shown that the osteogenic differentiation ability of BMSCs is significantly decreased in senile osteoporosis. Recently, circular RNAs (circRNAs) have been regarded as critical regulators in controlling the osteogenic differentiation of BMSCs by sponging microRNAs (miRNAs). Our study aimed to discover new and critical osteogenesis-related circRNAs that can promote bone formation in senile osteoporosis. METHODS: We detected the dysregulated circRNAs of BMSCs upon osteogenic differentiation induction and identified the critical osteogenic circRNA (circ-3626). The relationship between circ-3626 and osteoporosis was further verified in clinical bone samples and aged mice by qPCR. Moreover, circ-3626 AAV was constructed to examine the osteogenic effect of circ-3626 on bone formation via using Micro-CT, double calcein labeling, and the three-point bending tests. Bioinformatics analysis, Luciferase report gene assays, FISH, RNA pull-down, qPCR, Western Blots, and alizarin red staining assay explore the effects and mechanisms of circ-3626 on osteogenic differentiation of BMSCs. RESULTS: Circ-3626 was identified as a pivotal osteogenesis-related circRNA via RNA sequencing. The results of alizarin red staining, Western blots, and qPCR assays suggest that overexpressing circ-3626 dramatically accelerates the osteogenic capability of BMSCs. Furthermore, the bone repair capability of aging mice could be significantly improved by circ-3626 AAV treatment. Micro RNA miR-338-3p was identified as the downstream target of circ-3626. Overexpression of circ-3626 increases the expression of Runx2 by sponging miR-338-3p, thereby promoting the osteogenic differentiation of BMSCs by upregulating the expression of osteogenic genes. In addition, Western blots, and qPCR assays suggest circ-3626 AAV treatment promote the expression of Runx2 and osteogenic marker genes. CONCLUSION: Thus, we demonstrate that circ-3626 plays a pivotal role in promoting bone formation through the miR-338-3p/Runx2 axis and may provide new strategies for preventing and treating the bone loss of senile osteoporosis. CI - Copyright (c) 2023 The Author(s). Published by Elsevier Masson SAS.. All rights reserved. FAU - Chen, Gaoyang AU - Chen G AD - Division of Hand, Foot and Microvascular Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. Electronic address: chen199001207@163.com. FAU - Wang, Song AU - Wang S AD - Division of Spine Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. FAU - Wei, Ruihong AU - Wei R AD - Division of Hand, Foot and Microvascular Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. FAU - Liu, Yingnan AU - Liu Y AD - Division of Hand, Foot and Microvascular Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. FAU - Xu, Tao AU - Xu T AD - Division of Hand, Foot and Microvascular Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. FAU - Liu, Zhaokang AU - Liu Z AD - Division of Hand, Foot and Microvascular Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. FAU - Tan, Zhouyong AU - Tan Z AD - Division of Hand, Foot and Microvascular Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. FAU - Xie, Yongheng AU - Xie Y AD - Division of Spine Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. FAU - Yang, Dazhi AU - Yang D AD - Division of Spine Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. FAU - Liang, Zhen AU - Liang Z AD - Department of Geriatrics, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China. FAU - Zhuang, Yongqing AU - Zhuang Y AD - Division of Hand, Foot and Microvascular Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. Electronic address: zhuangyq_szhand@163.com. FAU - Peng, Songlin AU - Peng S AD - Division of Spine Surgery, Department of Orthopedic Surgery, Shenzhen People's Hospital (the Second Clinical Medical College of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology), 518020 Shenzhen, China; Key Laboratory of Musculoskeletal Tissue Reconstruction and Function Restoration (ZDSYS20200811143752005), Shenzhen Institute for Orthopedic Research, 518020 Shenzhen, China. Electronic address: dyffyy2@mail.sustech.edu.cn. LA - eng PT - Journal Article DEP - 20231130 PL - France TA - Joint Bone Spine JT - Joint bone spine JID - 100938016 RN - 0 (RNA, Circular) RN - 60MEW57T9G (alizarin) RN - 0 (MicroRNAs) RN - 0 (MIRN338 microRNA, human) RN - 0 (Runx2 protein, mouse) RN - 0 (Core Binding Factor Alpha 1 Subunit) RN - 0 (Anthraquinones) SB - IM MH - Humans MH - Mice MH - Animals MH - Osteogenesis/genetics MH - RNA, Circular/genetics/metabolism MH - *MicroRNAs/genetics/metabolism MH - *Osteoporosis/genetics MH - Cell Differentiation MH - Cells, Cultured MH - Core Binding Factor Alpha 1 Subunit/genetics MH - *Anthraquinones OTO - NOTNLM OT - Age-related osteoporosis OT - Bone formation OT - Bone marrow stromal cells OT - Circ-3626 OT - Runx2 OT - miR-338-3p EDAT- 2023/12/03 00:42 MHDA- 2024/03/01 06:44 CRDT- 2023/12/02 19:28 PHST- 2023/07/26 00:00 [received] PHST- 2023/10/11 00:00 [revised] PHST- 2023/11/09 00:00 [accepted] PHST- 2024/03/01 06:44 [medline] PHST- 2023/12/03 00:42 [pubmed] PHST- 2023/12/02 19:28 [entrez] AID - S1297-319X(23)00148-3 [pii] AID - 10.1016/j.jbspin.2023.105669 [doi] PST - ppublish SO - Joint Bone Spine. 2024 Mar;91(2):105669. doi: 10.1016/j.jbspin.2023.105669. Epub 2023 Nov 30.