Caspase-4/11 promotes hyperlipidemia and chronic kidney disease–accelerated vascular inflammation by enhancing trained immunity
Yu Sun, Yifan Lu, Lu Liu, Fatma Saaoud, Ying Shao, Keman Xu, Charles Drummer IV, Ramon Cueto, Huimin Shan, Xiaohua Jiang, Huaqing Zhao, Hong Wang, Xiaofeng Yang
Yu Sun, Yifan Lu, Lu Liu, Fatma Saaoud, Ying Shao, Keman Xu, Charles Drummer IV, Ramon Cueto, Huimin Shan, Xiaohua Jiang, Huaqing Zhao, Hong Wang, Xiaofeng Yang
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Research Article Inflammation Vascular biology

Caspase-4/11 promotes hyperlipidemia and chronic kidney disease–accelerated vascular inflammation by enhancing trained immunity

Abstract

To determine whether hyperlipidemia and chronic kidney disease (CKD) have a synergy in accelerating vascular inflammation via trained immunity (TI), we performed aortic pathological analysis and RNA-Seq of high-fat diet–fed (HFD-fed) 5/6 nephrectomy CKD (HFD+CKD) mice. We made the following findings: (a) HFD+CKD increased aortic cytosolic LPS levels, caspase-11 (CASP11) activation, and 998 gene expressions of TI pathways in the aorta (first-tier TI mechanism); (b) CASP11–/– decreased aortic neointima hyperplasia, aortic recruitment of macrophages, and casp11–gasdermin D–mediated cytokine secretion; (c) CASP11–/– decreased N-terminal gasdermin D (N-GSDMD) membrane expression on aortic endothelial cells and aortic IL-1B levels; (d) LPS transfection into human aortic endothelial cells resulted in CASP4 (human)/CASP11 (mouse) activation and increased N-GSDMD membrane expression; and (e) IL-1B served as the second-tier mechanism underlying HFD+CKD-promoted TI. Taken together, hyperlipidemia and CKD accelerated vascular inflammation by promoting 2-tier trained immunity.

Authors

Yu Sun, Yifan Lu, Lu Liu, Fatma Saaoud, Ying Shao, Keman Xu, Charles Drummer IV, Ramon Cueto, Huimin Shan, Xiaohua Jiang, Huaqing Zhao, Hong Wang, Xiaofeng Yang

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Figure 6

Cytosolic LPS is increased in the aorta of HFD+CKD and activates the casp4/11-GSDMD pathway.

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Cytosolic LPS is increased in the aorta of HFD+CKD and activates the cas...
(A) The expression of LPS endocytic protein HMGB1 was increased in the kidneys of patients with CKD. (B) The expression of HMGB1 was negatively correlated with GFR (GSE9493, GSE66494). (C and D) The LPS-FITC (2 μg/mL) was transfected into human aortic endothelial cell (HAECs) using FuGENE for 16 hours. The transfection efficiency was detected by flow cytometry (D) and verified by visualization with confocal microscopy images (C) (40× magnification). (E and F) HAECs were treated with blank control, direct LPS stimulation (2 μg/mL), LPS transfection (2 μg/mL), and transfection control. The activity of casp4 (E) and cleavage of N-GSDMD (F) were examined by FLICA and flow cytometry. (G) The secretion of IL-1β in the supernatant was determined by ELISA. (H) The design of a new competitive inhibitor of GSDMD cleavage. (I and J) The inhibition efficiency detection of GSDMD peptide inhibitor (4 μM) was examined by flow cytometry (n = 6). (K) The secretion of IL-1β via the N-GSDMD protein channel into the supernatant of LPS-transfected HAECs was detected in the presence and absence of GSDMD cleavage inhibitor by ELISA (n = 3). (L) The expression of adhesion molecule VCAM-1 was measured by flow cytometry in the LPD, LPS transfection, and LPS transfection after the addition of different concentrations of GSDMD inhibitor. Two-tailed Student’s t test was used in A. Pearson correlations were used in B. The Kruskal-Wallis test with Benjamini and Hochberg multiple-comparison method was used to control the overall FDR of 5% (D–G and J–L).