The mechanosensitive Piezo1 channels contribute to the arterial medial calcification

László Szabó; Norbert Balogh; Andrea Tóth; Ágnes Angyal; Mónika Gönczi; Dávid Máté Csiki; Csaba Tóth; Ildikó Balatoni; Viktória Jeney; László Csernoch; Beatrix Dienes

doi:10.3389/fphys.2022.1037230

The mechanosensitive Piezo1 channels contribute to the arterial medial calcification

Front Physiol. 2022 Nov 10:13:1037230. doi: 10.3389/fphys.2022.1037230. eCollection 2022.

Authors

László Szabó^{1

2

3}, Norbert Balogh¹, Andrea Tóth^{4

5}, Ágnes Angyal¹, Mónika Gönczi^{1

3}, Dávid Máté Csiki^{4

5}, Csaba Tóth⁶, Ildikó Balatoni⁷, Viktória Jeney⁴, László Csernoch^{1

3}, Beatrix Dienes¹

Affiliations

¹ Department of Physiology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
² Doctoral School of Molecular Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
³ ELKH-DE Cell Physiology Research Group, University of Debrecen, Debrecen, Hungary.
⁴ MTA-DE Lendület Vascular Pathophysiology Research Group, Research Centre for Molecular Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
⁵ Doctoral School of Molecular Cell and Immune Biology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
⁶ Department of Surgery, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
⁷ Clinical Center, University of Debrecen, Debrecen, Hungary.

Abstract

Vascular calcification (VC) is associated with a number of cardiovascular diseases, as well as chronic kidney disease. The role of smooth muscle cells (SMC) has already been widely explored in VC, as has the role of intracellular Ca²⁺ in regulating SMC function. Increased intracellular calcium concentration ([Ca²⁺]_i) in vascular SMC has been proposed to stimulate VC. However, the contribution of the non-selective Piezo1 mechanosensitive cation channels to the elevation of [Ca²⁺]_i, and consequently to the process of VC has never been examined. In this work the essential contribution of Piezo1 channels to arterial medial calcification is demonstrated. The presence of Piezo1 was proved on human aortic smooth muscle samples using immunohistochemistry. Quantitative PCR and Western blot analysis confirmed the expression of the channel on the human aortic smooth muscle cell line (HAoSMC). Functional measurements were done on HAoSMC under control and calcifying condition. Calcification was induced by supplementing the growth medium with inorganic phosphate (1.5 mmol/L, pH 7.4) and calcium (CaCl₂, 0.6 mmol/L) for 7 days. Measurement of [Ca²⁺]_i using fluorescent Fura-2 dye upon stimulation of Piezo1 channels (either by hypoosmolarity, or Yoda1) demonstrated significantly higher calcium transients in calcified as compared to control HAoSMCs. The expression of mechanosensitive Piezo1 channel is augmented in calcified arterial SMCs leading to a higher calcium influx upon stimulation. Activation of the channel by Yoda1 (10 μmol/L) enhanced calcification of HAoSMCs, while Dooku1, which antagonizes the effect of Yoda1, reduced this amplification. Application of Dooku1 alone inhibited the calcification. Knockdown of Piezo1 by siRNA suppressed the calcification evoked by Yoda1 under calcifying conditions. Our results demonstrate the pivotal role of Piezo1 channels in arterial medial calcification.

Keywords: Dooku1; Piezo1; Yoda1; calcification; intracellular calcium concentration; mechanosensation; vascular smooth muscle.