Long noncoding RNA repressor of adipogenesis negatively regulates the adipogenic differentiation of mesenchymal stem cells through the hnRNP A1‐PTX3‐ERK axis. Issue 7 (8th November 2020)

• Record Type: Journal Article
• Authors: Pan, Yiqian; Xie, Zhongyu; Cen, Shuizhong; Li, Ming; Liu, Wenjie; Tang, Su'an; Ye, Guiwen; Li, Jinteng; Zheng, Guan; Li, Zhaofeng; Yu, Wenhui; Wang, Peng; Wu, Yanfeng; Shen, Huiyong
• Is Part Of: Clinical and translational medicine. Volume 10:Issue 7(2020)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2020-11-08
• Subjects: adipogenesis -- long noncoding RNA -- mesenchymal stem cell -- tissue engineering; Clinical medicine -- Periodicals; Medicine, Experimental -- Periodicals; Medical innovations -- Periodicals; Molecular biology -- Periodicals; Pathology, Molecular -- Periodicals; 616.027
• DOI: 10.1002/ctm2.227
• Languages: English
• ISSNs: 2001-1326

Abstract

Abstract: Background: Mesenchymal stem cells (MSCs) are pluripotent stem cells that can differentiate via osteogenesis and adipogenesis. The mechanism underlying MSC lineage commitment still remains incompletely elucidated. Understanding the regulatory mechanism of MSC differentiation will help researchers induce MSCs toward specific lineages for clinical use. In this research, we intended to figure out the long noncoding RNA (lncRNA) that plays a central role in MSC fate determination and explore its application value in tissue engineering. Methods: The expression pattern of lncRNAs during MSC osteogenesis/adipogenesis was detected by microarray and qRT‐PCR. Lentivirus and siRNAs were constructed to regulate the expression of lncRNA repressor of adipogenesis ( ROA ). MSC osteogenesis/adipogenesis was evaluated by western blot and alizarin red/oil red staining. An adipokine array was used to select the paracrine/autocrine factor PTX3, followed by RNA interference or recombinant human protein stimulation to confirm its function. The activation of signaling pathways was also detected by western blot, and a small molecule inhibitor, SCH772984, was used to inhibit the activation of the ERK pathway. The interaction between ROA and hnRNP A1 was detected by RNA pull‐down and RIP assays. Luciferase reporter and chromatin immunoprecipitation assays were used to confirm the binding of hnRNP A1 to the PTX3 promotor. Additionally, an in vivo adipogenesis experiment was conducted to evaluate the regulatory value of ROA in tissue engineering. Results: In this study, we demonstrated that MSC adipogenesis is regulated by lncRNA ROA both in vitro and in vivo. Mechanistically, ROA inhibits MSC adipogenesis by downregulating the expression of the key autocrine/paracrine factor PTX3 and the downstream ERK pathway. This downregulation was achieved through transcription inhibition by impeding hnRNP A1 from binding to the promoter of PTX3 . Conclusions: ROA negatively regulates MSC adipogenesis through the hnRNP A1‐PTX3‐ERK axis. ROA may be an effective target for modulating MSCs in tissue engineering. Abstract : The expression of lncRNA ROA is significantly downregulated during MSC adipogenesis, thereby facilitating the binding of hnRNP A1 to the promoter of PTX3. Increased PTX3 activates the ERK1/2 pathway and promotes the adipogenic differentiation of MSCs in an autocrine/paracrine fashion. By modulating ROA, the efficiency of in vivo MSC adipogenesis can be effectively regulated.