1. Academic Validation
  2. VX-765 ameliorates renal injury and fibrosis in diabetes by regulating caspase-1-mediated pyroptosis and inflammation

VX-765 ameliorates renal injury and fibrosis in diabetes by regulating caspase-1-mediated pyroptosis and inflammation

  • J Diabetes Investig. 2022 Jan;13(1):22-33. doi: 10.1111/jdi.13660.
Si Wen 1 2 Fei Deng 3 Lulu Li 1 Li Xu 1 4 Xin Li 1 Qiuling Fan 1
Affiliations

Affiliations

  • 1 Department of Nephrology, First Hospital of China Medical University, Shenyang, China.
  • 2 Department of Nephrology, First Affiliated Hospital of Dalian Medical University, Dalian, China.
  • 3 Department of Urology, Second Xiangya Hospital of Central South University, Changsha, China.
  • 4 Department of Laboratory Medicine, First Hospital of China Medical University, Shenyang, China.
Abstract

Introduction: As a lytic inflammatory cell death, Pyroptosis has been recently described but has not been unequivocally elucidated in diabetic nephropathy (DN). VX-765 is a safe and effective inhibitor of Caspase-1, that was well tolerated in a phase II clinical trial in patients with epilepsy, but its application in DN is still undefined.

Materials and methods: Immunoblot, co-immunoprecipitation, confocal microscope and flow cytometry were used to analyze the effects of glucose on Pyroptosis in renal tubular epithelia (HK-2). In vitro, selective Caspase-1 inhibitors VX-765 and Z-YVAD-FMK were administered. Pyroptosis and fibrogenesis were determined by immunoblot, ELISA, cytotoxicity assay and flow cytometry. In vivo, diabetic mice were administered with 100 mg/kg VX-765. Renal function, pathological changes, and the expressions of NLRC4, GSDMD, IL-1β, collagen I, fibronectin and CD45 in renal cortex were evaluated.

Results: We identified NLRC4 as a sensor for Caspase-1 activation. Moreover, we provided morphological and molecular evidence for Pyroptosis in glucose-stressed tubular cells, including ballooned cell membrane, Caspase-1 immunoreactivity, GSDMD cleavage, and the release of inflammatory cytokine and cellular contents. All these effects were prevented by treatment with VX-765 or Z-YVAD-FMK, confirming that Caspase-1 effectively regulates the occurrence of Pyroptosis in HK-2 cells. In vivo, treatment of diabetic Animals with VX-765 ameliorated renal function, suppressed inflammatory cell infiltration and pyroptosis-associated protein expression, and mitigated tubulointerstitial fibrosis.

Conclusions: This work revealed that caspase-1-mediated Pyroptosis drives renal inflammation and fibrosis in diabetes. Our results are the first demonstration of VX-765 representing a promising therapeutic opportunity for alleviating the progression of DN.

Keywords

Caspase-1; Diabetic nephropathy; GSDMD; Inflammation; Pyroptosis; VX-765.

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