ALKBH5-Modified HMGB1-STING Activation Contributes to Radiation Induced Liver Disease via Innate Immune Response
ALKBH5-Modified HMGB1-STING Activation Contributes to Radiation Induced Liver Disease via Innate Immune Response
复制标题
ALKBH5 修饰的 HMGB1-STING 激活通过先天免疫反应导致辐射诱发的肝病
DOI：
10.1016/j.ijrobp.2021.05.115
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发表时间：
2021-08-30
影响因子：
7
通讯作者：
Zeng, Zhaochong
中科院分区：
文献类型：
--
作者：
Chen, Genwen;Zhao, Qianqian;Zeng, Zhaochong
目的：放射治疗对原发性肝癌的治疗至关重要，但不可避免地会造成肝损伤，这限制了其实施。N6-甲基腺苷（m6 A）甲基化参与许多分子功能。然而，它在放射性肝病（RILD）中的作用仍然未知。在此，我们研究m6 A甲基化在RILD.Methods和材料中的作用：采用甲基化RNA免疫沉淀测序和RNA转录组测序技术，研究了照射后人肝星状细胞（HSC）的甲基化模式。C3 H/HeN小鼠和干扰素基因刺激因子（STING）缺陷型小鼠接受分3次24戈伊的X射线照射。使用甲基化RNA免疫沉淀，RNA免疫沉淀，荧光素酶测定，和信使RNA衰变assay.Results：人肝星状细胞的高迁移率族蛋白1（HMGB 1）转录的m6 A甲基化进行了验证甲基化模式后8戈伊的X射线照射显示出显着差异。辐射募集AlkB同系物5（ALKBH 5）以使HMGB 1的3'非翻译区中的m6 A残基去甲基化，这导致STING-干扰素调节因子3信号传导的激活。ALKBH 5基因敲除后，HMGB 13 '端非翻译区转录水平发生变化，m6 A残基突变后，这种变化消失。引人注目的是，ALKBH 5缺乏或HMGB 1沉默都减弱了I型干扰素的产生并减少了肝细胞凋亡。ALKBH 5的体内消耗消除了HMGB 1介导的STING信号传导的上调，并减少了肝脏炎症，这与用辐射处理的STING（-/-）小鼠一致。值得注意的是，YTHDF 2（m6 A阅读蛋白）直接结合到HMGB 1 m6 A修饰的网站，并促进其degradation.Conclusions：ALKBH 5依赖性HMGB 1的表达介导的STING干扰素调节因子3在RILD的先天免疫反应。(C)2021爱思唯尔公司All rights reserved.
Purpose: Radiation therapy, which is vital for the treatment of primary liver cancer, comes with unavoidable liver injury, which limits its implementation. N6-methyladenosine (m6A) methylation is involved in many molecular functions. However, its role in radiation-induced liver diseases (RILD) remains unknown. Herein, we investigate the role of m6A methylation in RILD.Methods and Materials: Methylated RNA-immunoprecipitation sequencing and RNA transcriptome sequencing were used to reveal the methylation pattern of human hepatic stellate cells (HSCs) exposed to irradiation. C3H/HeN mice and stimulator of interferon genes (STING)-deficient mice underwent x-ray irradiation of 24 Gy in 3 fractions. The m6A methylation of the high-mobility group box 1 (HMGB1) transcript was validated using methylated RNA immunoprecipitation, RNA immuno-precipitation, luciferase assays, and a messenger RNA decay assay.Results: Human hepatic stellate cells showed significant differences in methylation patterns after 8 Gy of x-ray irradiation. Irradiation recruited AlkB homolog 5 (ALKBH5) to demethylate m6A residues in the 3' untranslated region of HMGB1, which resulted in the activation of STING-interferon regulatory factor 3 signaling. Changes in the transcription of the 3' untranslated region of HMGB1 occurred after the knockdown of ALKBH5, which were eliminated after m6A residue muta-tion. Strikingly, ALKBH5 deficiency or HMGB1 silencing both attenuated type I interferon production and decreased hepato-cyte apoptosis. In vivo depletion of ALKBH5 abolished the upregulation of HMGB1-mediated STING signaling and decreased liver inflammation, which was consistent with STING(-/-) mice treated with irradiation. Notably, YTHDF2 (m6A reader protein) directly bound to HMGB1 m6A-modified sites and promoted its degradation.Conclusions: ALKBH5-dependent HMGB1 expression mediates STING-interferon regulatory factor 3 innate immune response in RILD. (C) 2021 Elsevier Inc. All rights reserved.