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April 1, 2026British Journal of Pharmacology2 citationsOpen Access

Defining AV2‐1 as a novel pharmacological probe to target human and rodent TRPV2

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ALAndrea LeipeLeipzig UniversityJRJulia A. RoceretaTranslational Therapeutics (United States)RPRuth A. PumroyTranslational Therapeutics (United States)

Key Points

  • The objective is to identify and characterise a novel small-molecule activator of TRPV2 to advance understanding of its function in immune cells.
  • Screened a compound library using calcium imaging in HEK293 cells expressing mouse TRPV2
  • Validated lead compound AV2-1 through concentration-response analyses and microfluorometric assays
  • Conducted structural analysis with cryoEM to understand AV2-1 interaction with TRPV2
  • Performed mutagenesis to confirm binding site residues on TRPV2
  • Assessed efficacy in human macrophages using various imaging techniques
  • AV2-1 effectively activates TRPV2 across mouse, rat, and human orthologues
  • Binding analysis reveals stabilization of TRPV2's active conformation by AV2-1
  • Induced calcium signals and ionic currents were observed in human peripheral blood-derived macrophages
  • Confirmed specific interactions with TRPV2 residues His165 and Cys157
  • AV2-1 shows low toxicity while enhancing calcium signals in immune cells

Abstract

Abstract Background and Purpose Transient receptor potential vanilloid 2 (TRPV2) is a non‐selective cation channel implicated in immune cell functions. However, progress in understanding TRPV2 has been limited by a lack of potent and selective pharmacological tools, particularly those targeting the human variant. We aimed to identify and characterise a novel small‐molecule activator of TRPV2. Experimental Approach We screened a compound library using Ca 2+ imaging in HEK293 cells stably expressing mouse TRPV2. The lead compound AV2‐1 was validated by concentration–response analyses, microfluorometric Ca 2+ assays, and electrophysiological recordings. Structural insights were obtained from cryoEM of TRPV2 in complex with AV2‐1, and mutagenesis was performed to confirm binding site residues. The efficacy of AV2‐1 was assessed in human peripheral blood‐derived macrophages by Ca 2+ imaging, whole‐cell electrophysiology and TIRF microscopy to detect Ca 2+ microdomains. Key Results AV2‐1 is a novel TRPV2 activator showing robust efficacy across mouse, rat and human orthologues. Structural analysis reveals that AV2‐1 stabilises the channel in its active conformation by binding to an established intracellular pocket via TRPV2‐specific residues His165 and Cys157, as confirmed by mutagenesis experiments. AV2‐1 induces TRPV2‐dependent global Ca 2+ i signals, ionic currents and localised subplasmalemmal Ca 2+ microdomains in human peripheral blood‐derived macrophages. Conclusion and Implications AV2‐1 represents a novel pharmacological tool for probing TRPV2 function in immune cells, combining improved selectivity and potency with low toxicity. Its ability to activate human TRPV2 and elicit physiologically relevant Ca 2+ signals highlights its potential for advancing TRPV2 research and for therapeutic exploration in immune modulation and disease contexts.

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Cite This Study

Leipe et al. (2026) studied this question.

synapsesocial.com/papers/69cd7b345652765b073a90b8https://doi.org/10.1111/bph.70413
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