biostudies-literatureXian HNIDDK NIH HHSNIAID NIH HHSNCI NIH HHSNIAMS NIH HHS1370-1385.e8https://www.ebi.ac.uk/biostudies/studies/S-EPMC9378606biostudies-literatureUnknown55(8)Mitochondrial DNA (mtDNA) escaping stressed mitochondria provokes inflammation via cGAS-STING pathway activation and, when oxidized (Ox-mtDNA), it binds cytosolic NLRP3, thereby triggering inflammasome activation. However, it is unknown how and in which form Ox-mtDNA exits stressed mitochondria in non-apoptotic macrophages. We found that diverse NLRP3 inflammasome activators rapidly stimulated uniporter-mediated calcium uptake to open mitochondrial permeability transition pores (mPTP) and trigger VDAC oligomerization. This occurred independently of mtDNA or reactive oxygen species, which induce Ox-mtDNA generation. Within mitochondria, Ox-mtDNA was either repaired by DNA glycosylase OGG1 or cleaved by the endonuclease FEN1 to 500-650 bp fragments that exited mitochondria via mPTP- and VDACImmunityOxidized DNA fragments exit mitochondria via mPTP- and VDAC-dependent channels to activate NLRP3 inflammasome and interferon signaling.PMC9378606R00 DK100640P30 AR073761P30 CA014195K99 DK100640K01 AR077111U54 CA260591R25 AI147376P30 CA023100R37 AI043477R01 AR069876R01 AI043477Shadel GSOnyuru JHoffman HMOffenberger JWatari KXian HSampath HSanchez-Lopez EYing WKarin MfalseOxidized DNA fragments exit mitochondria via mPTP- and VDAC-dependent channels to activate NLRP3 inflammasome and interferon signaling.Mitochondrial DNA (mtDNA) escaping stressed mitochondria provokes inflammation via cGAS-STING pathway activation and, when oxidized (Ox-mtDNA), it binds cytosolic NLRP3, thereby triggering inflammasome activation. However, it is unknown how and in which form Ox-mtDNA exits stressed mitochondria in non-apoptotic macrophages. We found that diverse NLRP3 inflammasome activators rapidly stimulated uniporter-mediated calcium uptake to open mitochondrial permeability transition pores (mPTP) and trigger VDAC oligomerization. This occurred independently of mtDNA or reactive oxygen species, which induce Ox-mtDNA generation. Within mitochondria, Ox-mtDNA was either repaired by DNA glycosylase OGG1 or cleaved by the endonuclease FEN1 to 500-650 bp fragments that exited mitochondria via mPTP- and VDAC2022-01-01T00:00:00Z2022 Aug2026-05-28T02:15:58.229Z2026-04-08T02:07:57.23ZS-EPMC93786063583510710.1016/j.immuni.2022.06.007