Mycobacterial EAST-6 triggers cGAS-STING pathway via the GSDMD-NTdependent mitochondria dysfunction

ESAT-6 induces mitochondrial dysfunction, characterized by mitochondrial DNA (mtDNA) release and mitochondrial ROS (mtROS) production. Further investigation revealed that ESAT-6 activates the NLRP3 inflammasome and GSDMD cleavage, followed by and formation of GSDMD-NT. GSDMD-NT then translocates to mitochondria, disrupting their integrity and promoting mtDNA and mtROS release. GSDMD deficiency prevented ΔΨm loss, mtDNA/mtROS release, and IFN-β responses. Therefore, our findings establish a novel ESAT-6-GSDMD-mtDNA axis that drives type I IFN responses, providing critical insights into Mtb immune evasion strategies.