Targeted metabolomics analysis by LC-MS/MS for BMDCs infected with VSV

Virus-infected cells often exhibit dramatic cellular changes accompanied by altered mitochondrial function. The contribution of factors shaping the inner mitochondrial membrane (IMM) and cristae architecture on viral replication is insufficiently understood. Single cell transcriptomics applying vesicular stomatitis virus (VSV) infection suggests involvement of factors determining IMM architecture following infection. Consistently, inhibition of the F1Fo ATP synthase reduces viral replication which is associated with oligomerization of the complex and altered IMM structure. Moreover, deletion of mitochondrial contact site and cristae organizing system (MICOS) complex by targeting MIC60 results in reduced viral replication. Generation of Mic60inv/inv CD11c-Cre+ mice uncovers reduced crista junctions and viral replication in bone marrow-derived dendritic cells. Consequently, reduced viral replication in CD11c-expressing cells limits prolonged immune activation. Altogether, by linking the F1Fo ATP synthase and the MICOS complex to viral replication and immune activation, we describe links between the mitochondrial structure-metabolism and the immune response against viral infection.

病毒感染的细胞往往会出现显著的细胞变化，并伴随线粒体功能异常。目前对于调控线粒体内膜（inner mitochondrial membrane, IMM）及嵴结构的因子在病毒复制过程中的贡献，尚缺乏充分认知。应用水疱性口炎病毒（vesicular stomatitis virus, VSV）感染开展的单细胞转录组学研究提示，病毒感染后调控线粒体内膜结构的相关因子参与了感染进程。与此一致的是，抑制F1Fo ATP合酶可降低病毒复制水平，该效应与该复合物的寡聚化及线粒体内膜结构改变密切相关。此外，通过靶向MIC60敲除线粒体接触位点和嵴组装复合物（mitochondrial contact site and cristae organizing system, MICOS），同样可使病毒复制能力受损。构建Mic60inv/inv CD11c-Cre+小鼠后发现，其骨髓来源树突状细胞中的嵴连接数量减少，且病毒复制水平显著降低。由此，表达CD11c的细胞中病毒复制的减弱会限制持续的免疫激活。综上，本研究将F1Fo ATP合酶与MICOS复合物关联至病毒复制及免疫激活过程，阐明了线粒体结构-代谢与抗病毒免疫应答之间的内在联系。