LncRNA GAS5 inhibits Th17 differentiation and alleviates immune thrombocytopenia via promoting the ubiquitination of STAT3
LncRNA GAS5 inhibits Th17 differentiation and alleviates immune thrombocytopenia via promoting the ubiquitination of STAT3
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LncRNA GAS5通过促进STAT3泛素化抑制Th17分化并缓解免疫性血小板减少症
DOI：
10.1016/j.intimp.2019.106127
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发表时间：
2020-03-01
影响因子：
5.6
通讯作者：
Xia, Yalin
中科院分区：
文献类型：
--
作者：
Li, Jianqin;Tian, Jianmei;Xia, Yalin
背景资料：辅助性T细胞17（Th 17）的分化增加加速了免疫性血小板减少症（ITP）的发展，这是一种常见的自身免疫性疾病，治疗方法有限。研究表明，长链非编码RNA（IncRNA）在自身免疫性疾病中发挥重要作用，本研究旨在探讨IncRNA GAS 5对ITP患者Th 17细胞分化的影响。流式细胞仪检测Th 17细胞在CD 44（+）细胞中的比例。通过RNA下拉实验和RNA结合蛋白免疫沉淀（RIP）证实GASS和STAT 3之间的结合。泛素化实验检测STAT 3的泛素化，免疫共沉淀（Co-IP）检测STAT 3与TRAF 6的相互作用。结果：GAS 5在ITP患者外周血单个核细胞（PBMC）和ITP小鼠脾组织中表达下调。GAS 5过表达抑制了幼稚CD 4（+）细胞中Th 17的分化，但对Treg的分化没有影响。RNA pull-down和RNA免疫沉淀分析证实了GAS 5和STAT 3之间的相互作用。进一步的研究表明GAS 5通过促进TRAF 6介导的泛素化而加速STAT 3的降解。结论：LncRNA GAS 5通过促进TRAF 6介导的STAT 3泛素化，抑制Th 17细胞分化，从而减轻ITP。
Background: The increased differentiation of T helper 17 cells (Th17) accelerates the development of immune thrombocytopenia (ITP), which is a common autoimmune disease with limited therapeutic methods. Recent studies have revealed that long non-coding RNAs (IncRNAs) play a critical role in autoimmune diseases, thus this study aims to investigate the effect of IncRNA GAS5 on the differentiation of Th17 cells in ITP.Methods: The expression of GAS5 in peripheral blood mononuclear cells (PBMCs) of ITP patients and spleen tissues of ITP mice was measured by qRT-PCR. The percentage of Th17 cells in CD44(+) cells was measured by flow cytometry. The combination between GASS and STAT3 was confirmed by RNA pull-down assay and RNA Binding Protein Immunoprecipitation (RIP). The ubiquitination of STAT3 was detected by ubiquitination assay and the interaction between STAT3 and TRAF6 was measured by Co-Immunoprecipitation (Co-IP). Finally, the effect of GASS on Th17 differentiation was investigated in vitro and in vivo using lentivirus (lenti)-GAS5.Results: GAS5 expression was downregulated both in PBMCs of ITP patients and spleen tissues of ITP mice. Overexpression of GAS5 suppressed Th17 differentiation while had no effect on Treg differentiation in naive CD4(+) cells. RNA pull-down and RNA immunoprecipitation assays confirmed the interaction between GAS5 and STAT3. Further studies showed GAS5 accelerated the degradation of STAT3 via promoting TRAF6-mediated ubiquitination. Overexpressing GAS5 suppressed Th17 differentiation in vitro and alleviated ITP in vivo via reducing STAT3.Conclusion: LncRNA GAS5 inhibited Th17 differentiation through promoting the TRAF6-mediated ubiquitination of STAT3, thus relieving ITP.