Caspase 3/GSDME-dependent pyroptosis contributes to chemotherapy drug-induced nephrotoxicity

Xiujin Shen,Hong Jiang,Haibing Wang,Chunhua Weng,Jianghua Chen

doi:10.1038/s41419-021-03458-5

Abstract

Chemotherapy drug-induced nephrotoxicity limits clinical applications for treating cancers. Pyroptosis, a newly discovered programmed cell death, was recently reported to be associated with kidney diseases. However, the role of pyroptosis in chemotherapeutic drug-induced nephrotoxicity has not been fully clarified. Herein, we demonstrate that the chemotherapeutic drug cisplatin or doxorubicin, induces the cleavage of gasdermin E (GSDME) in cultured human renal tubular epithelial cells, in a time- and concentration-dependent manner. Morphologically, cisplatin- or doxorubicin-treated renal tubular epithelial cells exhibit large bubbles emerging from the cell membrane. Furthermore, activation of caspase 3, not caspase 9, is associated with GSDME cleavage in cisplatin- or doxorubicin-treated renal tubular epithelial cells. Meanwhile, silencing GSDME alleviates cisplatin- or doxorubicin-induced HK-2 cell pyroptosis by increasing cell viability and decreasing LDH release. In addition, treatment with Ac-DMLD-CMK, a polypeptide targeting mouse caspase 3-Gsdme signaling, inhibits caspase 3 and Gsdme activation, alleviates the deterioration of kidney function, attenuates renal tubular epithelial cell injury, and reduces inflammatory cytokine secretion in vivo. Specifically, GSDME cleavage depends on ERK and JNK signaling. NAC, a reactive oxygen species (ROS) inhibitor, reduces GSDME cleavage through JNK signaling in human renal tubular epithelial cells. Thus, we speculate that renal tubular epithelial cell pyroptosis induced by chemotherapy drugs is mediated by ROS-JNK-caspase 3-GSDME signaling, implying that therapies targeting GSDME may prove efficacious in overcoming chemotherapeutic drug-induced nephrotoxicity.

Highlights

Traditional chemotherapeutic drugs, such as cisplatin and doxorubicin, are commonly used to treat various cancers, including lung, bladder, and ovarian cancer[1,2,3,4]
Cisplatin or doxorubicin induces pyroptosis of renal tubular epithelial cells It has been reported that the typical characteristics of pyroptosis were increased LDH release, increased PIpositive cells with flow cytometry, and typical bubbles emerging from the cell membrane[14]
Flow cytometry analysis demonstrated that cisplatin dramatically increased the proportion of propidium iodide (PI)+positive cells in a concentration-dependent manner (Fig. S1e, f) Morphologically, both the cisplatin- and doxorubicin-treated HK-2 cells showed typical bubbles emerging from the cell membrane (Fig. S1g)

Summary

Introduction

Traditional chemotherapeutic drugs, such as cisplatin and doxorubicin, are commonly used to treat various cancers, including lung, bladder, and ovarian cancer[1,2,3,4]. Severe side effects caused by toxicity to healthy organs and tissues, the kidney, limit the clinical application of these drugs[5,6]. Pyroptosis is a newly discovered form of programmed cell death with morphological characteristics that differ. Pyroptosis can be induced by activation of the executors, gasdermin E (GSDME), or gasdermin D (GSDMD), which results in the cleavage of their N-terminal fragments (GSDME-N or GSDMD-N, respectively)[13,14,15]. GSDME-N or GSDMD-N translocate to the cell membrane and mediate cell perforation, resulting in infiltration of extracellular material, cell swelling, and pyroptosis[16]. Moderate cell pyroptosis can remove pathogenic microorganisms and antagonize infection, excessive cell pyroptosis leads to cell death and enhances inflammatory responses, resulting in fever, hypotension, septicemia, as well as other serious symptoms[12]

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