Phosphate burden induces vascular calcification through a NLRP3-caspase-1-mediated pyroptotic pathway

Li-Chun Ho; Yu-Hsin Chen; Ting-Yun Wu; Ling-Zhen Kao; Shih-Yuan Hung; Hung-Hsiang Liou; Pei-Chun Chen; Pei-Jane Tsai; Hui-Kuan Lin; Yi-Che Lee; Hsi-Hao Wang; Yau-Sheng Tsai

doi:10.1016/j.lfs.2023.122123

Phosphate burden induces vascular calcification through a NLRP3-caspase-1-mediated pyroptotic pathway

Life Sci. 2023 Nov 1:332:122123. doi: 10.1016/j.lfs.2023.122123. Epub 2023 Sep 22.

Authors

Li-Chun Ho¹, Yu-Hsin Chen², Ting-Yun Wu³, Ling-Zhen Kao³, Shih-Yuan Hung⁴, Hung-Hsiang Liou⁵, Pei-Chun Chen⁶, Pei-Jane Tsai⁷, Hui-Kuan Lin⁸, Yi-Che Lee⁹, Hsi-Hao Wang⁴, Yau-Sheng Tsai¹⁰

Affiliations

¹ School of Medicine, College of Medicine, I-Shou University, Kaohsiung, Taiwan; Division of General Medicine, Department of Internal Medicine, E-Da Hospital, I-Shou University, Kaohsiung, Taiwan. Electronic address: hoim7024@ms3.hinet.net.
² Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan; Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
³ Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
⁴ Division of Nephrology, Department of Internal Medicine, E-Da Hospital, I-Shou University, Kaohsiung, Taiwan.
⁵ Division of Nephrology, Department of Internal Medicine, Hsin-Jen Hospital, New Taipei City, Taiwan.
⁶ Department of Physiology, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
⁷ Department of Medical Laboratory Science and Biotechnology, College of Medicine, National Cheng Kung University, Tainan, Taiwan; Department of Cancer Biology, Wake Forest Baptist Medical Center, Wake Forest University, Winston Salem, NC, USA.
⁸ Department of Cancer Biology, Wake Forest Baptist Medical Center, Wake Forest University, Winston Salem, NC, USA.
⁹ School of Medicine, College of Medicine, I-Shou University, Kaohsiung, Taiwan.
¹⁰ Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan; Department of Cancer Biology, Wake Forest Baptist Medical Center, Wake Forest University, Winston Salem, NC, USA; Clinical Medicine Research Center, National Cheng Kung University Hospital, Tainan, Taiwan. Electronic address: yaustsai@mail.ncku.edu.tw.

PMID: 37742736
DOI: 10.1016/j.lfs.2023.122123

Abstract

Aims: The aim of this study is to clarify the role of NLRP3 inflammasome in phosphate burden-induced vascular smooth muscle cell (VSMC) calcification.

Main methods: VSMC calcification was induced using a high concentration of inorganic phosphate. After pharmacological inhibition or genetic silencing of the NLRP3 inflammasome, pyroptosis, or potassium efflux, the cells were examined by RT-qPCR, immunofluorescence, and western blotting to identify the NLRP3-mediated pathway for VSMC calcification.

Key findings: Calcified VSMCs with α-smooth muscle actin (α-SMA) disarray presented features of pyroptosis, including caspase-1 maturation, cleaved gasdermin D (GSDMD), and a high supernatant level of lactate dehydrogenase A. Pharmacological inhibitions of caspase-1 and pyroptosis attenuated VSMC calcification, whereas interleukin-1β receptor antagonism did not. Unlike canonical NLRP3 activation, osteogenic VSMCs did not upregulate NLRP3 expression. However, NLRP3 genetic silencing or inhibitions, which targets different domains of the NLRP3 protein, could ameliorate VSMC calcification by aborting caspase-1 and GSDMD activation. Furthermore, potassium efflux through the inward-rectifier potassium channel, and not through the P2X7 receptor, triggered NLRP3 inflammasome activation and VSMC calcification.

Significance: In the present study, we identified a potassium efflux-triggered NLRP3-caspase-1-mediated pyroptotic pathway for VSMC calcification that is unique and different from the canonical NLRP3 inflammasome activation. Therefore, targeting this pathway may serve as a novel therapeutic strategy for vascular calcification.

Keywords: NLRP3 inflammasome; Pyroptosis; Vascular calcification.