CHMP4B and VSP4A reverse GSDMD-mediated pyroptosis by cell membrane remodeling in endometrial carcinoma

Ye Yang; Hai-Lian Chen; Su Fang Wu; Wei Bao

doi:10.1016/j.bbagen.2023.130497

CHMP4B and VSP4A reverse GSDMD-mediated pyroptosis by cell membrane remodeling in endometrial carcinoma

Biochim Biophys Acta Gen Subj. 2024 Jan;1868(1):130497. doi: 10.1016/j.bbagen.2023.130497. Epub 2023 Nov 4.

Authors

Ye Yang¹, Hai-Lian Chen², Su Fang Wu³, Wei Bao⁴

Affiliations

¹ Obstetrics and Gynecology Department, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 85 Wujin Road, Hongkou, Shanghai 200080, PR China.
² Surgical Department, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 85 Wujin Road, Hongkou, Shanghai 200080, PR China.
³ Obstetrics and Gynecology Department, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 85 Wujin Road, Hongkou, Shanghai 200080, PR China. Electronic address: wusufang73@163.com.
⁴ Obstetrics and Gynecology Department, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 85 Wujin Road, Hongkou, Shanghai 200080, PR China. Electronic address: forever_chipper@163.com.

PMID: 37931722
DOI: 10.1016/j.bbagen.2023.130497

Abstract

Background: In advanced and recurrent endometrial carcinoma (EC), the current state of immuno- or targeted therapy remains in the clinical research phase. Our study aimed to explore the role of the ESCRT machinery in maintaining cell membrane integrity and reversing pyroptotic cell death.

Methods: Immunohistochemistry, western blotting, and co-immunoprecipitation were performed to determine the expression and relationship between GSDMD, CHMP4B, and VPS4A. We employed techniques such as FITC Annexin V/propidium iodide staining, Ca²⁺ fluorescence intensity, IL-1β enzyme-linked immunosorbent assay, and lactate dehydrogenase release assay to detect pyroptosis in endometrial cancer cells. Plasma membrane perforations and CHMP4B/VPS4A puncta were observed through electron and fluorescence confocal microscopy.

Results: We showed that GSDMD, CHMP4B, and VPS4A were differentially expressed in the pyroptotic EC xenograft mouse model group, as well as high, moderate, and mild expression in EC cells treated with LPS and nigericin compared to endometrial epithelial cells. Co-IP confirmed the interaction between GSDMD, CHMP4B, and VPS4A. We found that GSDMD knockdown reduced PI-positive cells, Ca²⁺ efflux, IL-1β, and LDH release, while CHMP4B and VPS4A depletion enhanced these indicators in HEC1A and AN3CA cells. Electron microscopy showed membrane perforations correspondingly decreased with inactivated GSDMD and increased or decreased after CHMP4B and VPS4A depletion or overexpression in EC cells. Fluorescence confocal microscopy detected CHMP4B protein puncta associated with VPS4A at the injured plasma membrane in GSDMD^NT cells.

Conclusions: We preliminary evidenced that CHMP4B and VPS4A reverses GSDMD-mediated pyroptosis by facilitating cell membrane remodeling in endometrial carcinoma. Targeting CHMP4B related proteins may promote pyroptosis in endometrial tumors.

Keywords: CHMP4B; Cell membrane remodeling; Endometrial carcinoma; GSDMD; Pyroptosis; VPS4A.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ATPases Associated with Diverse Cellular Activities / metabolism
Animals
Cell Membrane / metabolism
Disease Models, Animal
Endometrial Neoplasms* / metabolism
Endosomal Sorting Complexes Required for Transport / metabolism
Female
Gasdermins
Humans
Intracellular Signaling Peptides and Proteins / metabolism
Mice
Phosphate-Binding Proteins / metabolism
Pyroptosis
Vacuolar Proton-Translocating ATPases* / metabolism

Substances

Intracellular Signaling Peptides and Proteins
CHMP4B protein, human
Endosomal Sorting Complexes Required for Transport
VPS4A protein, human
ATPases Associated with Diverse Cellular Activities
Vacuolar Proton-Translocating ATPases
GSDMD protein, human
Gasdermins
Phosphate-Binding Proteins
CHMP4B protein, mouse