lncRNA H19 mediates BMP9-induced osteogenic differentiation of mesenchymal stem cells (MSCs) through Notch signaling.

// Junyi Liao 1, 2 , Xinyi Yu 1, 2 , Xue Hu 1, 2 , Jiaming Fan 2, 3 , Jing Wang 1, 2 , Zhicai Zhang 2, 4 , Chen Zhao 1, 2 , Zongyue Zeng 2, 3 , Yi Shu 2, 3 , Ruyi Zhang 2, 3 , Shujuan Yan 2, 3 , Yasha Li 2, 3 , Wenwen Zhang 2, 5 , Jing Cui 2, 3 , Chao Ma 2, 6 , Li Li 2, 7 , Yichun Yu 2, 8 , Tingting Wu 2, 6 , Xingye Wu 1, 2 , Jiayan Lei 1, 2 , Jia Wang 2, 3 , Chao Yang 2, 3 , Ke Wu 2, 3 , Ying Wu 2, 9 , Jun Tang 10 , Bai-Cheng He 2, 3 , Zhong-Liang Deng 2, 3 , Hue H. Luu 2 , Rex C. Haydon 2 , Russell R. Reid 2, 11 , Michael J. Lee 2 , Jennifer Moriatis Wolf 2 , Wei Huang 1 and Tong-Chuan He 2, 3 1 Departments of Orthopaedic Surgery, Blood Transfusion, Nephrology, and General Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China 2 Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, IL, USA 3 Ministry of Education Key Laboratory of Diagnostic Medicine, and The Affiliated Hospitals of Chongqing Medical University, Chongqing, China 4 Department of Orthopaedic Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science & Technology, Wuhan, China 5 Department of Laboratory Medicine and Clinical Diagnostics, The Affiliated Yantai Hospital, Binzhou Medical University, Yantai, China 6 Departments of Neurosurgery, and Otolaryngology-Head & Neck Surgery, The Affiliated Zhongnan Hospital of Wuhan University, Wuhan, China 7 Department of Biomedical Engineering, School of Bioengineering, Chongqing University, Chongqing, China 8 Department of Emergency Medicine, Beijing Hospital, Beijing, China 9 Department of Immunology and Microbiology, Beijing University of Chinese Medicine, Beijing, China 10 Cytate Institute for Precision Medicine & Innovation, Guangzhou Cytate Biomedical Technologies Inc., Guangzhou, China 11 Department of Surgery, Section of Plastic Surgery, The University of Chicago Medical Center, Chicago, IL, USA Correspondence to: Tong-Chuan He, email: tche@uchicago.edu Wei Huang, email: huangwei68@263.net Keywords: mesenchymal stem cells, BMP9, osteogenic differentiation, lncRNA H19, Notch signaling Received: March 16, 2017      Accepted: May 23, 2017      Published: June 27, 2017 ABSTRACT Mesenchymal stem cells (MSCs) are multipotent progenitor cells that can undergo self-renewal and differentiate into multiple lineages. Osteogenic differentiation from MSCs is a well-orchestrated process and regulated by multiple signaling pathways. We previously demonstrated that BMP9 is one of the most potent osteogenic factors. However, molecular mechanism through which BMP9 governs osteoblastic differentiation remains to be fully understood. Increasing evidence indicates noncoding RNAs (ncRNAs) may play important regulatory roles in many physiological and/or pathologic processes. In this study, we investigate the role of lncRNA H19 in BMP9-regulated osteogenic differentiation of MSCs. We demonstrated that H19 was sharply upregulated at the early stage of BMP9 stimulation of MSCs, followed by a rapid decease and gradual return to basal level. This process was correlated with BMP9-induced expression of osteogenic markers. Interestingly, either constitutive H19 expression or silencing H19 expression in MSCs significantly impaired BMP9-induced osteogenic differentiation in vitro and in vivo , which was effectively rescued by the activation of Notch signaling. Either constitutive H19 expression or silencing H19 expression led to the increased expression of a group of miRNAs that are predicted to target Notch ligands and receptors. Thus, these results indicate that lncRNA H19 functions as an important mediator of BMP9 signaling by modulating Notch signaling-targeting miRNAs. Our findings suggest that the well-coordinated biphasic expression of lncRNA H19 may be essential in BMP9-induced osteogenic differentiation of MSCs, and that dysregulated H19 expression may impair normal osteogenesis, leading to pathogenic processes, such as bone tumor development.