KS0365, a novel activator of the transient receptor potential vanilloid 3 (TRPV3) channel, accelerates keratinocyte migration

Abstract

Background and Purpose: Ca 2+ signalling mediated by the thermosensitive, nonselective, Ca 2+ -permeable transient receptor potential channel TRPV3 is assumed to play a critical role in regulating several aspects of skin functions, such as keratinocyte proliferation, differentiation, skin barrier formation and wound healing. Studying the function of TRPV3 in skin homeostasis, however, is still constrained by a lack of potent and selective pharmacological modulators of TRPV3.Experimental Approach: By screening an in-house compound library using fluorometric intracellular Ca 2+ assays, we identified two chemically related hits. The more potent and efficient TRPV3 activator 2-(2-chloro-3-isopropylcyclopent-2-en-1-yl)-4-methylphenol (KS0365) was further evaluated in fluo-4-assisted Ca 2+ assays, different Ca 2+ imaging approaches, electrophysiological studies, cytotoxicity and migration assays. Key Results: KS0365 activated recombinant and native mouse TRPV3 more potently and with a higher efficacy compared with 2-APB and did not activate TRPV2 or TRPV4 channels. The activation of TRPV3 by KS0365 super-additively accelerated the EGF-induced keratinocyte migration, which was inhibited by the TRP channel blocker ruthenium red or by siRNA-mediated TRPV3 knockdown. Moreover, KS0365 induced strong Ca 2+ responses in migrating front cells and in leading edges of keratinocytes. Conclusions and Implications: The selective TRPV3 activator KS0365 triggers increases in [Ca 2+ ] i with most prominent signals in the leading edge and accelerates migration of keratinocytes. TRPV3 activators may promote re-epithelialization upon skin wounding.

TRPV3 Ion Channel: From Gene to Pharmacology

“…Other data indicated that TRPV3 regulates nitric oxide synthesis in the skin, thus promoting wound healing in vivo [153]. A synthetic activator KS0365 accelerated keratinocyte migration that paralleled strong TRPV3-mediated calcium responses in migrating front cells and in leading edges, which suggests the role of TRPV3 in re-epithelialization upon skin wounding [54].…”

“…A novel synthetic activator of TRPV3, KS0365 showed a higher efficacy and potency than 2-APB. The compound also acted non-selectively, acting on TRPV1 and TRPV2 channels and triggering intracellular calcium release at higher concentrations [54].…”

Multimodal roles of transient receptor potential channel activation in inducing pathological tissue scarification

“…TRPV3, on the other hand, can induce fibrosis through TRPV3/TSLP/Smad2/3 pathway, resulting in significantly increased expression levels of α-SMA, fibronectin, COL1A1, and TSLP ( 62 ). Additionally, the selective TRPV3 activator KS0365 has been shown to accelerate the migration of KCs and promote re-epithelialization during physiological wound healing ( 63 ). Furthermore, the presence of TRPV3 in macrophage lysosomes may play a crucial role in the inflammatory phase of PS ( 64 ).…”

“…Studies have shown that the new TRPV3 channel activator KS0365 promotes wound healing by accelerating the re-epithelialization of KCs, while the broad-spectrum channel blocker ruthenium red and siRNA-mediated TRPV3 knockdown inhibit this process ( 63 , 159 ). This finding indicates that overexpression of TRPV3 channels can promote excessive wound healing leading to the formation of PS.…”

Novel Insights into the Role of Keratinocytes-Expressed TRPV3 in the Skin

“…For example, TRPV3 is a promising target for improving wound healing. In the epidermis, appropriately-activated TRPV3 can promote keratinocyte proliferation, differentiation, and migration and inhibit cell apoptosis, thus accelerating re-epithelialization [ 76 , 94 ]. In the dermis, TRPV3 is involved in the process of dermal extracellular matrix deposition, which can improve the integrity of injured skin, which also suggests the potential of TRPV3 in regulating scar formation [ 95 ].…”