N6-methyladenosine demethylase FTO regulates inflammatory cytokine secretion and tight junctions in retinal pigment epithelium cells

作者全名："Tang, Shiyun; Meng, Jiayu; Tan, Jun; Liu, Xianyang; Zhou, Hongxiu; Li, Na; Hou, Shengping"

作者地址："[Tang, Shiyun; Meng, Jiayu; Tan, Jun; Liu, Xianyang; Zhou, Hongxiu; Hou, Shengping] Chongqing Med Univ, Affiliated Hosp 1, Chongqing, Peoples R China; [Tang, Shiyun; Meng, Jiayu; Tan, Jun; Liu, Xianyang; Zhou, Hongxiu; Hou, Shengping] Chongqing Key Lab Ophthalmol, Chongqing, Peoples R China; [Tang, Shiyun; Meng, Jiayu; Tan, Jun; Liu, Xianyang; Zhou, Hongxiu; Hou, Shengping] Chongqing Eye Inst, Chongqing, Peoples R China; [Tang, Shiyun; Meng, Jiayu; Tan, Jun; Liu, Xianyang; Zhou, Hongxiu; Hou, Shengping] Natl Clin Res Ctr Ocular Dis, Chongqing Branch, Chongqing, Peoples R China; [Li, Na] Chongqing Med Univ, Coll Basic Med, Chongqing, Peoples R China; [Hou, Shengping] Chongqing Med Univ, Affiliated Hosp 1, Chongqing 400016, Peoples R China"

通信作者："Hou, SP (通讯作者)，Chongqing Med Univ, Affiliated Hosp 1, Chongqing, Peoples R China.; Hou, SP (通讯作者)，Chongqing Key Lab Ophthalmol, Chongqing, Peoples R China.; Hou, SP (通讯作者)，Chongqing Eye Inst, Chongqing, Peoples R China.; Hou, SP (通讯作者)，Natl Clin Res Ctr Ocular Dis, Chongqing Branch, Chongqing, Peoples R China.; Li, N (通讯作者)，Chongqing Med Univ, Coll Basic Med, Chongqing, Peoples R China.; Hou, SP (通讯作者)，Chongqing Med Univ, Affiliated Hosp 1, Chongqing 400016, Peoples R China."

来源：CLINICAL IMMUNOLOGY

ESI学科分类：IMMUNOLOGY

WOS号：WOS:000841551100001

JCR分区：Q1

影响因子：8.6

年份：2022

卷号：241

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：N6-methyladenosine; FTO; Uveitis; Experimental autoimmune uveitis; Retinal pigment epithelium

摘要："Objective: Uveitis is an intraocular inflammatory disease. Epigenetics has been associated with its pathogenesis. However, the role of N6-methyladenosine (m6A) in uveitis has not been reported. We aimed to examine the role of m6A and its regulatory mechanism in experimental autoimmune uveitis (EAU).Methods: The mRNA expression of m6A-related methylase and demethylase of retinal pigment epithelium (RPE) between mice with EAU and control mice was detected by RT-qPCR. The overall m6A level of ARPE-19 cells was detected by an m6A quantitative detection kit. Cell proliferation was observed by CCK-8 assays, and ELISA was used to test the secretion of inflammatory factors. The expression of tight junction proteins and the target genes of FTO were examined by western blotting and MeRIP-PCR.Results: A decreased expression of FTO in RPE cells was found in mice with EAU. Increased overall m6A%, proliferation of cells and secretion of IL-6, IL-8 and MCP-1 were found after FTO knockdown in ARPE-19 cells. However, ZO-1 and occludin protein expression was decreased. ATF4 protein expression was decreased in the FTO knockdown (shFTO) group as compared with the control (shNC) group. In contrast, the m6A level of ATF4 was elevated, as shown by MeRIP-PCR. Functional analysis showed that p-STAT3 expression was increased in the shFTO group, and the change in occludin expression was reversed in ATF4 rescue experiment.Conclusion: FTO may affect the translation of ATF4 by regulating its m6A level, resulting in the increased expression of p-STAT3 and inflammatory factors, and leading to uveitis."

基金机构："National Natural Science Foundation Project of China [82070951, 81873678]; Chongqing Education Commission [CXQT19015, W0047]; Natural Science Foundation Project of Chongqing [cstc2019jcyjmsxmX0120]; Plan of Overseas Study of Chongqing [cx2018010]; Chongqing Education Commission [KJQN202000406]; National Key Clinical Specialties Construction Program of China, Chongqing Branch of National Clinical Research Center for Ocular Diseases, Chongqing Key Laboratory of Ophthalmology (CSTC) [KJQN202000406]; Natural Science Foundation Project of Chongqing Medical University [2008CA5003]"

基金资助正文："This study was supported by National Natural Science Foundation Project of China (82070951, 81873678) , The Innovative Research Group Project of Chongqing Education Commission (CXQT19015) , Natural Science Foundation Project of Chongqing (cstc2019jcyjmsxmX0120) , the innovation supporting Plan of Overseas Study of Chongqing (cx2018010) , the Grant of Chongqing Education Commission (KJQN202000406) and National Key Clinical Specialties Construction Program of China, Chongqing Branch of National Clinical Research Center for Ocular Diseases, Chongqing Key Laboratory of Ophthalmology (CSTC, 2008CA5003) and Natural Science Foundation Project of Chongqing Medical University (W0047) ."