H19 Increases IL-17A/IL-23 Releases via Regulating VDR by Interacting with miR675-5p/miR22-5p in Ankylosing Spondylitis

Xu Zhang; Shuang Ji; Guoqi Cai; Zhipeng Pan; Renfang Han; Yaping Yuan; Shanshan Xu; Jiajia Yang; Xingxing Hu; Mengya Chen; Meng Wu; Yubo Ma; Jixiang Deng; Xing Gao; Shiyang Guan; Shengqian Xu; Zongwen Shuai; Laura Laslett; Faming Pan

doi:10.1016/j.omtn.2019.11.025

H19 Increases IL-17A/IL-23 Releases via Regulating VDR by Interacting with miR675-5p/miR22-5p in Ankylosing Spondylitis

Mol Ther Nucleic Acids. 2020 Mar 6:19:393-404. doi: 10.1016/j.omtn.2019.11.025. Epub 2019 Nov 29.

Authors

Xu Zhang¹, Shuang Ji², Guoqi Cai¹, Zhipeng Pan³, Renfang Han⁴, Yaping Yuan⁴, Shanshan Xu⁴, Jiajia Yang⁴, Xingxing Hu⁴, Mengya Chen⁴, Meng Wu⁴, Yubo Ma⁴, Jixiang Deng⁴, Xing Gao⁴, Shiyang Guan⁴, Shengqian Xu⁵, Zongwen Shuai⁵, Laura Laslett⁶, Faming Pan⁷

Affiliations

¹ Department of Epidemiology and Biostatistics, School of Public Health, Anhui Medical University, Hefei, China; Menzies Institute for Medical Research, University of Tasmania, Hobart, Australia; The Key Laboratory of Major Autoimmune Diseases, Anhui Medical University, Hefei, China.
² Department of Respiratory and Critical Care Medicine, First Affiliated Hospital of Anhui Medical University, Hefei, China.
³ Department of Medical Oncology, First Affiliated Hospital of Anhui Medical University, Hefei, China.
⁴ Department of Epidemiology and Biostatistics, School of Public Health, Anhui Medical University, Hefei, China; The Key Laboratory of Major Autoimmune Diseases, Anhui Medical University, Hefei, China.
⁵ Department of Rheumatism and Immunity, First Affiliated Hospital of Anhui Medical University, Hefei, China.
⁶ Menzies Institute for Medical Research, University of Tasmania, Hobart, Australia.
⁷ Department of Epidemiology and Biostatistics, School of Public Health, Anhui Medical University, Hefei, China; Menzies Institute for Medical Research, University of Tasmania, Hobart, Australia; The Key Laboratory of Major Autoimmune Diseases, Anhui Medical University, Hefei, China. Electronic address: famingpan@ahmu.edu.cn.

Abstract

Long non-coding RNA (lncRNA) H19 is associated with inflammatory diseases, but the molecular mechanism of H19 in the inflammatory process of ankylosing spondylitis (AS) is unclear. Here, we investigated the role of H19 and its downstream molecules in the inflammation of AS by microarray analysis, qRT-PCR, western blot, and dual-luciferase reporter assay. H19 small interfering RNA (siRNA) (Si-H19) and adenovirus (AD-H19) were used to decrease and increase H19 expression, respectively. 42 annotated lncRNAs were identified, and H19 was overexpressed. H19, vitamin D receptor (VDR), and transforming growth factor β (TGF-β) can bind to microRNA22-5p (miR22-5p) and miR675-5p. Si-H19 significantly downregulated miR22-5p and upregulated miR675-5p expression; Si-H19 decreased the protein and mRNA expression of VDR and decreased the cytokine and mRNA levels of interleukin-17A (IL-17A) and IL-23. These results were verified by AD-H19. In addition, miR22-5p and miR675-5p inhibitors increased the protein and mRNA expression of VDR and increased the cytokine and mRNA levels of IL-17A and IL-23. These results were also confirmed by miRNA mimics. Furthermore, H19 directly interfered with miR22-5p and miR675-5p expression, whereas the two miRNAs directly inhibited VDR expression. Overall, the H19-miR22-5p/miR675-5p-VDR-IL-17A/IL-23 signaling pathways have important roles in the pathogenesis of AS.

Keywords: H19; IL-17A; IL-23; VDR; ankylosing spondylitis; cytokines; miR22; miR675; non-coding RNA.