Liu Tao, Hecker Julian, Liu Siqi, Rui Xianliang, Boyer Nathan, Wang Jennifer, Yu Yuzhen, Zhang Yihan, Mou Hongmei, Gomez-Escobar Luis Guillermo, Choi Augustine M K, Raby Benjamin A, Weiss Scott T, Zhou Xiaobo
Channing Division of Network Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
The Mucosal Immunology and Biology Research Center, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02115, USA.
J Respir Biol Transl Med. 2024 Mar;1(1). doi: 10.35534/jrbtm.2024.10005. Epub 2024 Mar 31.
Released mitochondrial DNA (mtDNA) in cells activates cGAS-STING pathway, which induces expression of interferon-stimulated genes (ISGs) and thereby promotes inflammation, as frequently seen in asthmatic airways. However, whether the genetic determinant, Gasdermin B (GSDMB), the most replicated asthma risk gene, regulates this pathway remains unknown. We set out to determine whether and how GSDMB regulates mtDNA-activated cGAS-STING pathway and subsequent induction in human airway epithelial cells. Using qPCR, ELISA, native polyacrylamide gel electrophoresis, co-immunoprecipitation and immunofluorescence assays, we evaluated the regulation of GSDMB on cGAS-STING pathway in both BEAS-2B cells and primary normal human bronchial epithelial cells (nHBEs). mtDNA was extracted in plasma samples from human asthmatics and the correlation between mtDNA levels and eosinophil counts was analyzed. is significantly associated with expression in asthmatic nasal epithelial brushing samples from the Genes-environments and Admixture in Latino Americans (GALA) II study. Over-expression of promotes DNA-induced IFN and in bronchial epithelial BEAS-2B cells and nHBEs. Conversely, knockout of led to weakened induction of (IFNs) and in BEAS-2B cells. Mechanistically, GSDMB interacts with the C-terminus of STING, promoting the translocation of STING to Golgi, leading to the phosphorylation of IRF3 and induction of and . mtDNA copy number in serum from asthmatics was significantly correlated with blood eosinophil counts especially in male subjects. GSDMB promotes the activation of mtDNA and poly (dA:dT)-induced activation of cGAS-STING pathway in airway epithelial cells, leading to enhanced induction of
细胞中释放的线粒体DNA（mtDNA）激活cGAS-STING通路，该通路诱导干扰素刺激基因（ISGs）的表达，从而促进炎症，这在哮喘气道中很常见。然而，遗传决定因素Gasdermin B（GSDMB），即最易复制的哮喘风险基因，是否调节该通路仍不清楚。我们着手确定GSDMB是否以及如何调节mtDNA激活的cGAS-STING通路以及随后在人呼吸道上皮细胞中的诱导作用。我们使用qPCR、ELISA、天然聚丙烯酰胺凝胶电泳、免疫共沉淀和免疫荧光测定法，评估了GSDMB对BEAS-2B细胞和原代正常人支气管上皮细胞（nHBEs）中cGAS-STING通路的调节作用。从人类哮喘患者的血浆样本中提取mtDNA，并分析mtDNA水平与嗜酸性粒细胞计数之间的相关性。在拉丁裔美国人基因-环境与混合（GALA）II研究的哮喘鼻上皮刷检样本中，[具体内容缺失]与[具体内容缺失]表达显著相关。[具体内容缺失]的过表达促进支气管上皮BEAS-2B细胞和nHBEs中DNA诱导的IFN和[具体内容缺失]。相反，[具体内容缺失]的敲除导致BEAS-2B细胞中[具体内容缺失]（IFNs）和[具体内容缺失]的诱导减弱。机制上，GSDMB与STING的C末端相互作用，促进STING向高尔基体的转运，导致IRF3的磷酸化以及[具体内容缺失]和[具体内容缺失]的诱导。哮喘患者血清中的mtDNA拷贝数与血液嗜酸性粒细胞计数显著相关，尤其是在男性受试者中。GSDMB促进气道上皮细胞中mtDNA的激活以及聚（dA:dT）诱导的cGAS-STING通路的激活，导致[具体内容缺失]的诱导增强
