Shear stress activates ADAM10 sheddase to regulate Notch1 via the Piezo1 force sensor in endothelial cells.

Vincenza Caolo,T Simon Futers,Laeticia Lichtenstein,Naima Endesh,Gregory Parsonage,Fiona Bartoli,David J Beech,Elizabeth Av Jones,Marjolaine Debant

doi:10.7554/elife.50684

Vincenza Caolo, T Simon Futers + Show 7 more

Open Access

https://doi.org/10.7554/elife.50684

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Journal: eLife	Publication Date: Jun 2, 2020
Citations: 55	License type: CC BY 4.0

Affiliation: University of Leeds

Abstract

Mechanical force is a determinant of Notch signalling but the mechanism of force detection and its coupling to Notch are unclear. We propose a role for Piezo1 channels, which are mechanically-activated non-selective cation channels. In cultured microvascular endothelial cells, Piezo1 channel activation by either shear stress or a chemical agonist Yoda1 activated a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10), a Ca2+-regulated transmembrane sheddase that mediates S2 Notch1 cleavage. Consistent with this observation, we found Piezo1-dependent increase in the abundance of Notch1 intracellular domain (NICD) that depended on ADAM10 and the downstream S3 cleavage enzyme, γ-secretase. Conditional endothelial-specific disruption of Piezo1 in adult mice suppressed the expression of multiple Notch1 target genes in hepatic vasculature, suggesting constitutive functional importance in vivo. The data suggest that Piezo1 is a mechanism conferring force sensitivity on ADAM10 and Notch1 with downstream consequences for sustained activation of Notch1 target genes and potentially other processes.

Highlights

Mammalian Notch proteins were identified following studies in D. melanogaster that linked genetic abnormality to wing notch (Siebel and Lendahl, 2017)
Statistical analysis indicated no significant change in this basal Notch1 intracellular domain (NICD) signal (Figure 1b), its existence in some individual experiments complicated our determination of whether shear stress induced an increase in NICD (Figure 1a,b)
The study has identified a connection between Piezo1 channels and Notch1 signalling and a novel mechanism by which Notch1 can be regulated and impacted by mechanical force

Summary

Introduction

Mammalian Notch proteins were identified following studies in D. melanogaster that linked genetic abnormality to wing notch (Siebel and Lendahl, 2017). Piezo channels are Ca2+-permeable non-selective cationic channels, so when force causes them to open there is Ca2+ entry, elevation of the cytosolic Ca2+ concentration and regulation of Ca2+dependent mechanisms (Coste et al, 2010; Murthy et al, 2017) Relevant to such a system is Ca2+ and Ca2+-calmodulin regulation of ADAM10 (Nagano et al, 2004; Maretzky et al, 2015), a metalloprotease or sheddase that catalyses rate-limiting S2 cleavage of Notch prior to gsecretase-mediated S3 cleavage and release of Notch intracellular domain (NICD), driving downstream transcription (Siebel and Lendahl, 2017; Alabi et al, 2018; Anders et al, 2001). We focussed on endothelial cells where both proteins are prominent and have established functional significance (Siebel and Lendahl, 2017; Murthy et al, 2017; Li et al, 2014; Rode et al, 2017; Wu et al, 2017; Alabi et al, 2018)

Results

Discussion

Materials and methods