Yoda1 analogue (Dooku1) which antagonizes Yoda1‐evoked activation of Piezo1 and aortic relaxation

Abstract

Background and PurposeThe mechanosensitive Piezo1 channel has important roles in vascular physiology and disease. Yoda1 is a small‐molecule agonist, but the pharmacology of these channels is otherwise limited.Experimental ApproachYoda1 analogues were generated by synthetic chemistry. Intracellular Ca2+ and Tl+ measurements were made in HEK 293 or CHO cell lines overexpressing channel subunits and in HUVECs, which natively express Piezo1. Isometric tension recordings were made from rings of mouse thoracic aorta.Key ResultsModification of the pyrazine ring of Yoda1 yielded an analogue, which lacked agonist activity but reversibly antagonized Yoda1. The analogue is referred to as Dooku1. Dooku1 inhibited 2 μM Yoda1‐induced Ca2+‐entry with IC50s of 1.3 μM (HEK 293 cells) and 1.5 μM (HUVECs) yet failed to inhibit constitutive Piezo1 channel activity. It had no effect on endogenous ATP‐evoked Ca2+ elevation or store‐operated Ca2+ entry in HEK 293 cells or Ca2+ entry through TRPV4 or TRPC4 channels overexpressed in CHO and HEK 293 cells. Yoda1 caused dose‐dependent relaxation of aortic rings, which was mediated by an endothelium‐ and NO‐dependent mechanism and which was antagonized by Dooku1 and analogues of Dooku1.Conclusion and ImplicationsChemical antagonism of Yoda1‐evoked Piezo1 channel activity is possible, and the existence of a specific chemical interaction site is suggested with distinct binding and efficacy domains.

Inhibition of Piezo1 attenuates demyelination in the central nervous system

“…Yoda-1 (Tocris, 5586) is a synthetic small molecule that specifically activates Piezo1 at micromolar concentrations (Evans et al, 2018;Syeda et al, 2015) by interacting with the agonist transduction motif of Piezo1 subunits, thus enhancing channel opening-time (Lacroix, Botello-Smith, & Luo, 2018). In contrast to GsMTx4, Yoda-1 may interact directly with Piezo1 domains rather than modifying the lipid environment around the ion channel (Syeda et al, 2015).…”

Sphingomyelinase disables Piezo1 channel inactivation to enable sustained response to mechanical force

“…We first studied cultured human umbilical vein endothelial cells (HUVECs) as a model of native endothelium. These cells express functional Piezo1 channels 11,31 . A synthetic chemical agonist, Yoda1 31,32 , activated the channels to cause Ca 2+ entry (Figure 2a).…”

“…These cells express functional Piezo1 channels 11,31 . A synthetic chemical agonist, Yoda1 31,32 , activated the channels to cause Ca 2+ entry (Figure 2a). The response to Yoda1 was suppressed by Gd 3+ , a non-specific Piezo1 channel blocker 14 or by depletion of Piezo1 caused by Piezo1-targeted siRNA, consistent with Piezo1 channels mediating the response (Figure 2a Neutral sphingomyelinase inhibitors prevent sustained endothelial response to flow To investigate flow responses of native channels, endothelium was freshly isolated from second-order mesenteric arteries of adult mice.…”

Piezo1 channel activates ADAM10 sheddase to regulate Notch1 and gene expression

“…Therefore, we speculated about a potential relationship between Piezo1 and Notch1, using endothelial cells to investigate this idea because both proteins are prominent in these cells and have established functional significance in them 1,[8][9][10]12,19 . Because mechanical force can affect numerous mechanisms, we explored the relationship by specifically activating Piezo1 channels with a synthetic small-molecule agonist (Yoda1) that acts directly to enhance force sensitivity [21][22][23][24] . Because there is inherent force in cell membranes and force arises through cell-cell and cell-substrate contact, Yoda1 can be used to activate the channels in endothelial cells without applying an exogenous force 24 that might otherwise complicate the analysis by concurrently stimulating parallel mechanisms.…”