Caspase-12 exhibits non-redundant functions in response to endoplasmic reticulum stress to promote GSDMD-mediated NETosis, leading to thoracic aortic dissection

Article

作者: Ma, Xiang ; Yang, Kun ; Ge, Junbo ; Wei, Wei ; Gao, Pingjin ; Li, Yulin ; Ling, Yunzhi ; Yesitayi, Gulinazi ; Sun, Aijun ; Zhang, Shumin ; Lyu, Yang ; Chen, Hanchuan ; Gao, Rifeng ; Shi, Jiaran

BACKGROUNDThoracic aortic dissection (TAD) is a highly lethal condition that is characterized by inflammatory cell infiltration. Recent evidence has indicated that Gasdermin D (GSDMD) plays an important role in vascular inflammation and degeneration. However, its effects on neutrophil extracellular trap formation and release (NETosis) during TAD remain unknown.METHODSA TAD mouse model was generated using four-week-old male neutrophil-specific GSDMD-knockout mice (GSDMD^F/F; Elane^Cre) and dimethyl fumarate (DMF)-treated C57BL/6J mice by administering β-aminopropionitrile monofumarate (BAPN; 1 g/kg/day) in their drinking water for 4 weeks. Immunoprecipitation and immunofluorescence assays were performed to examine the role of the endoplasmic reticulum (ER) and its associated protein, caspase-12, in GSDMD-induced NETosis.RESULTSGSDMD was elevated and co-localized primarily in neutrophils in the aortic tissues of patients with TAD and mice with BAPN-induced TAD. This was accompanied by increased NETosis. Neutrophil-specific GSDMD knockout and the NETosis inhibitor, GSK484, mitigated TAD development in mice. However, GSK484 did not provide additional therapeutic effects against TAD in the neutrophil-specific, GSDMD knockout mice. Mechanistically, ER stress promoted GSDMD cleavage by caspase-4/11, thereby inducing NETosis. Furthermore, caspase-12 exhibited non-redundant functions in the cleavage of GSDMD by caspase-4/11. The GSDMD inhibitor, DMF, partially prevented TAD development.CONCLUSIONSThe ER stress/GSDMD/NETosis signaling pathway provides a potential therapeutic target for the prevention and treatment of TAD.

2025-03-01·Cellular Signalling

Aquaporin-5 facilitates liver regeneration following hepatectomy via ROS/GSDMD pathway

Article

作者: Han, Wenshuo ; Chen, Peng ; Di, Guohu ; Li, Bin ; Sun, Hetong ; Wang, Dianqiang ; Ge, Huanhuan ; Song, Peirong ; Wang, Ye

During the proliferative phase of liver regeneration, insufficient regulation of hepatocyte hydrogen peroxide (H₂O₂) overproduction can result in oxidative stress and hepatocyte death. This study aims to investigate the influence of Aquaporin 5 (Aqp5) on liver regeneration by evaluating its role in reactive oxygen species (ROS) generation and NLRP3-GSDMD-mediated pyroptosis. A 70 % partial hepatectomy (PHx) model was established in Aqp5^-/- mice to evaluate the pathological changes in the liver. Reactive oxygen species (ROS) production was assessed using a dichlorodihydrofluorescein diacetate (DCFH-DA) assay. Aqp5 deficiency significantly increased ROS production, the number of TUNEL-positive cells, and disrupted mitochondrial membrane potential in the liver of Aqp5-deficient mice. The impact of Aqp5 on ROS/NLRP3/Gasdermin-D (GSDMD)-mediated pyroptosis was examined through the administration of N-acetyl-L-cysteine (NAC, an ROS scavenger) or disulfiram (DSF, a GSDMD inhibitor). In Aqp5-deficient mice, the regenerative liver exhibited increased expression of NLRP3, enhanced activation of caspase-1 and GSDMD, as well as elevated secretion of IL-1β. Treatment with DSF significantly attenuated GSDMD-mediated pyroptosis triggered by Aqp5 deficiency in the regenerating liver. Furthermore, the administration of NAC to Aqp5-deficient mice resulted in a reduction in the expression levels of NLRP3, the activity levels of caspase-1 and GSDMD, as well as the release of IL-1β. Our findings indicate that the deficiency of Aqp5 facilitates GSDMD activation through the production of ROS. The suppression of ROS or inhibition of GSDMD significantly alleviates the damage and pyroptosis observed in Aqp5-deficient regenerative liver.

2025-03-01·Clinical and Translational Medicine

The novel GSDMD inhibitor GI‐Y2 exerts antipyroptotic effects to reduce atherosclerosis

Article

作者: Fan, Xiaoxi ; Ye, Keke ; Tang, Zhixuan ; Cheng, Zhenfeng ; Huang, Weijian ; Chen, Xudong ; Ye, Bozhi ; Dai, Shanshan ; Shi, Si ; Cai, Xueli ; Shao, Ruiyin ; Han, Jibo ; Zheng, Wenyuan

AbstractIntroductionGasdermin D (GSDMD) and the pyroptosis it mediates are importantly involved in cardiovascular diseases (CVDs). Identifying and developing new inhibitors of GSDMD could be a promising strategy for treating pyroptosis‐mediated diseases, such as atherosclerosis.ObjectivesWe aimed to develop new inhibitor of GSDMD in atherosclerosis, as well as clarify the mechanisms underlying this inhibiting effect.MethodsSurface plasmon resonance and pull‐down assay were used to identify the amino acid sites of GSDMD inhibited by GI‐Y2. A mouse model of atherosclerosis was established by feeding a high‐fat diet for 12 weeks. After treating mice with GI‐Y2 (10 or 20 mg/kg, i.g.), the lipid plaque area on the arterial intimal surface, lipid deposition, collagen deposition and pyroptosis levels in aortic root sections were evaluated. Additionally, further treatment of atherosclerotic mice with macrophage membrane‐encapsulated GI‐Y2 was conducted to enhance the targeting ability of GI‐Y2 to atherosclerotic plaques.ResultsIn this study, we confirmed GI‐Y2 as a novel inhibitor of GSDMD via structure‐based virtual screening and pharmacological validation. Mechanistically, GI‐Y2 directly interacts with the Arg10 residue of GSDMD and reduces the membrane binding of GSDMD‐N. Functionally, we revealed that GI‐Y2 inhibits the formation of atherosclerotic plaques by targeting GSDMD. Similarly, GI‐Y2 reduces pyroptosis and macrophage infiltration in atherosclerosis. Furthermore, we constructed macrophage membrane‐coated GI‐Y2 nanoparticles to enhance the targeting of GI‐Y2 to macrophages in atheromatous plaques and demonstrated its vascular protective effect in vivo.ConclusionThis work demonstrated that GI‐Y2 can potentially alleviate CVDs by targeting GSDMD and provided a new compound for the study of GSDMD‐mediated pyroptosis.Key points

We preliminarily confirmed GI‐Y2 as a novel inhibitor of GSDMD via structure‐based virtual screening and pharmacological validation.

GI‐Y2 directly interacts with GSDMD and reduces the membrane binding of GSDMD‐N via the Arg10 residue.

GI‐Y2 inhibits the formation of atherosclerotic plaques by targeting GSDMD and GI‐Y2 reduces pyroptosis and macrophage infiltration in atherosclerosis.

We constructed macrophage membrane‐coated GI‐Y2 nanoparticles to enhance the targeting of GI‐Y2 to macrophages in atheromatous plaques and demonstrated its vascular protective effect in vivo.

100 项与 GSDMD 抑制剂 (炎明生物) 相关的药物交易

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