Gasdermin D (GSDMD)-activated inflammatory cell death (pyroptosis) causes mitochondrial damage, but its underlying mechanism and functional consequences are largely unknown. Here, we show that the N-terminal pore-forming GSDMD fragment (GSDMD-NT) rapidly damaged both inner and outer mitochondrial membranes (OMMs) leading to reduced mitochondrial numbers, mitophagy, ROS, loss of transmembrane potential, attenuated oxidative phosphorylation (OXPHOS), and release of mitochondrial proteins and DNA from the matrix and intermembrane space. Mitochondrial damage occurred as soon as GSDMD was cleaved prior to plasma membrane damage. Mitochondrial damage was independent of the B-cell lymphoma 2 family and depended on GSDMD-NT binding to cardiolipin. Canonical and noncanonical inflammasome activation of mitochondrial damage, pyroptosis, and inflammatory cytokine release were suppressed by genetic ablation of cardiolipin synthase (Crls1) or the scramblase (Plscr3) that transfers cardiolipin to the OMM. Phospholipid scramblase-3 (PLSCR3) deficiency in a tumor compromised pyroptosis-triggered anti-tumor immunity. Thus, mitochondrial damage plays a critical role in pyroptosis.

Gasdermin D （GSDMD） 激活的炎症细胞死亡（焦亡）会导致线粒体损伤，但其潜在机制和功能后果在很大程度上尚不清楚。在这里，我们表明 N 末端成孔 GSDMD 片段 （GSDMD-NT） 迅速破坏线粒体内膜和外膜 （OMM），导致线粒体数量减少、线粒体自噬、ROS、跨膜电位丧失、氧化磷酸化减弱 （OXPHOS），以及线粒体蛋白和 DNA 从基质和膜间空间释放。一旦 GSDMD 在质膜损伤之前被切割，就会发生线粒体损伤。线粒体损伤独立于 B 细胞淋巴瘤 2 家族，并依赖于 GSDMD-NT 与心磷脂的结合。通过将心磷脂合成酶 （Crls1） 或将心磷脂转移到 OMM 的加扰酶 （Plscr3） 的基因消融抑制线粒体损伤、焦亡和炎性细胞因子释放的经典和非经典炎性小体激活。磷脂扰乱酶 3 （PLSCR3） 缺陷在肿瘤中损害了焦亡触发的抗肿瘤免疫。因此，线粒体损伤在焦亡中起着关键作用。