Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death
Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death
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DOI：
10.1038/nature15514
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发表时间：
2015-10-29
期刊：
影响因子：
64.8
通讯作者：
Shao, Feng
中科院分区：
文献类型：
--
作者：
Shi, Jianjin;Zhao, Yue;Shao, Feng
炎性半胱天冬酶（半胱天冬酶-1、-4、-5和-11）对于先天性防御至关重要。Caspase-1被各种典型炎性小体的配体激活，caspase-4、-5和-11直接识别细菌脂多糖，这两者都触发细胞凋亡。尽管在免疫和内毒素休克中起关键作用，但炎性半胱天冬酶诱导的细胞凋亡机制尚不清楚。在这里，我们通过在小鼠骨髓巨噬细胞中caspase-11和caspase-1介导的焦亡的全基因组成簇规则间隔回文重复（CRISPR）-Cas9核酸酶筛选来鉴定gasdermin D（Gsdmd）。GSDMD缺陷细胞抵抗胞质脂多糖和已知的典型炎性体配体诱导的细胞凋亡。白细胞介素-1 β的释放也减少Gsdmd（-/-）细胞，尽管完整的加工半胱天冬酶-1。Caspase-1和Caspase-4/5/11特异性切割GSDMD中氨基末端gasdermin-N和羧基末端gasdermin-C结构域之间的连接体，这是焦亡所必需的并且是足够的。裂解释放的gasdermin-N结构域上的分子内抑制，表现出内在的pyroptosis-inducing活性。其他gasdermin家族成员不被炎性半胱天冬酶切割，但共享自抑制; Gsdma 3中导致脱发和皮肤缺陷的功能获得性突变破坏了自抑制，使其gasdermin-N结构域触发焦亡。这些发现提供了对炎性小体介导的免疫/疾病的深入了解，也改变了我们对焦亡和程序性坏死的理解。
Inflammatory caspases (caspase-1, -4, -5 and -11) are critical for innate defences. Caspase-1 is activated by ligands of various canonical inflammasomes, and caspase-4, -5 and -11 directly recognize bacterial lipopolysaccharide, both of which trigger pyroptosis. Despite the crucial role in immunity and endotoxic shock, the mechanism for pyroptosis induction by inflammatory caspases is unknown. Here we identify gasdermin D (Gsdmd) by genome-wide clustered regularly interspaced palindromic repeat (CRISPR)-Cas9 nuclease screens of caspase-11- and caspase-1-mediated pyroptosis in mouse bone marrow macrophages. GSDMD-deficient cells resisted the induction of pyroptosis by cytosolic lipopolysaccharide and known canonical inflammasome ligands. Interleukin-1 beta release was also diminished in Gsdmd(-/-) cells, despite intact processing by caspase-1. Caspase-1 and caspase-4/5/11 specifically cleaved the linker between the amino-terminal gasdermin-N and carboxy-terminal gasdermin-C domains in GSDMD, which was required and sufficient for pyroptosis. The cleavage released the intramolecular inhibition on the gasdermin-N domain that showed intrinsic pyroptosis-inducing activity. Other gasdermin family members were not cleaved by inflammatory caspases but shared the autoinhibition; gain-of-function mutations in Gsdma3 that cause alopecia and skin defects disrupted the autoinhibition, allowing its gasdermin-N domain to trigger pyroptosis. These findings offer insight into inflammasome-mediated immunity/diseases and also change our understanding of pyroptosis and programmed necrosis.