IRF2‐mediated upregulation of lncRNA HHAS1 facilitates the osteogenic differentiation of bone marrow‐derived mesenchymal stem cells by acting as a competing endogenous RNA. Issue 6 (20th June 2021)

• Record Type: Journal Article
• Authors: Ye, Guiwen; Wang, Peng; Xie, Zhongyu; Li, Jinteng; Zheng, Guan; Liu, Wenjie; Cao, Qian; Li, Ming; Cen, Shuizhong; Li, Zhaofeng; Yu, Wenhui; Wu, Yanfeng; Shen, Huiyong
• Is Part Of: Clinical and translational medicine. Volume 11:Issue 6(2021)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-06-20
• Subjects: IRF2 -- long noncoding RNAs -- mesenchymal stem cells -- osteogenic differentiation; Clinical medicine -- Periodicals; Medicine, Experimental -- Periodicals; Medical innovations -- Periodicals; Molecular biology -- Periodicals; Pathology, Molecular -- Periodicals; 616.027
• DOI: 10.1002/ctm2.429
• Languages: English
• ISSNs: 2001-1326

Abstract

Abstract: Background: Mesenchymal stem cells (MSCs) are the major source of osteoblasts. Long noncoding RNAs (lncRNAs) are abundantly expressed RNAs that lack protein‐coding potential and play an extensive regulatory role in cellular biological activities. However, the regulatory network of lncRNAs in MSC osteogenesis needs further investigation. Methods: QRT‐PCR, western blot, immunofluorescence, and immunohistochemistry assays were used to determine the levels of relevant genes. The osteogenic differentiation capability was evaluated by using Alizarin Red S (ARS) staining, alkaline phosphatase activity assays, hematoxylin & eosin staining or micro‐CT. RNA fluorescence in situ hybridization (FISH) and RNAscope were used to detect HHAS1 expression in cells and bone tissue. A microarray assay was performed to identify differentially expressed microRNAs. RNA immunoprecipitation and RNA pull‐down were used to explore the interactions between related proteins and nucleic acids. Results: The level of lncRNA HHAS1 increased during bone marrow‐derived MSC (BMSC) osteogenesis and was positively related to the levels of osteogenic genes and ARS intensity. HHAS1 was located in both the cytoplasm and the nucleus and was expressed in human bone tissue. HHAS1 facilitated BMSC osteogenic differentiation by downregulating miR‐204‐5p expression and enhancing the level of RUNX family transcription factor 2 (RUNX2). In addition, interferon regulatory factor 2 (IRF2) was increased during BMSC osteogenic differentiation and interacted with the promoter of HHAS1, which resulted in the transcriptional activation of HHAS1. Furthermore, IRF2 and HHAS1 helped improve bone defect repair in vivo. Conclusions: Our study identified a novel lncRNA, HHAS1, that facilitates BMSC osteogenic differentiation and proposed a role for the IRF2/HHAS1/miR‐204‐5p/RUNX2 axis in BMSC osteogenesis regulation. These findings help elucidate the regulatory network of BMSC osteogenesis and provide potential targets for clinical application. Abstract : Graphical Highlights 1. Under the condition of osteogenic induction, the level of IRF2 is increased and promotes the transcription of lncRNA HHAS1. 2. HHAS1 sponges miR‐204‐5p by acting as a ceRNA, thereby protecting RUNX2 mRNA from translation inhibition and facilitating BMSC osteogenesis. 3. Modulating IRF2 or HHAS1 can regulate BMSC osteogenic differentiation and help improve bone defect impair.