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

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.

• Authors: Si Wen, Fei Deng, Lulu Li, Li Xu, Xin Li, Qiuling Fan
• Journal: Journal of diabetes investigation (J Diabetes Investig) Vol. 13 Issue 1 Pg. 22-33 (Jan 2022) ISSN: 2040-1124 [Electronic] Japan
• PMID: 34494385 (Publication Type: Journal Article)
• Copyright: © 2021 The Authors. Journal of Diabetes Investigation published by Asian Association for the Study of Diabetes (AASD) and John Wiley & Sons Australia, Ltd.

Chemical References

• Dipeptides
• para-Aminobenzoates
• belnacasan
• Caspase 1
• Glucose

Topics

• Animals
• Caspase 1 (metabolism)
• Cell Culture Techniques
• Diabetes Mellitus, Experimental (drug therapy, pathology)
• Diabetic Nephropathies (drug therapy, pathology)
• Dipeptides (pharmacology)
• Fibrosis
• Glucose (pharmacology)
• Humans
• Inflammation
• Kidney (pathology)
• Kidney Tubules (drug effects, pathology)
• Male
• Mice
• Pyroptosis (drug effects)
• para-Aminobenzoates (pharmacology)