A human TRPV1 genetic variant within the channel gating domain regulates pain sensitivity in rodents
Shufang He, … , Ye Zhang, Eric R. Gross
Shufang He, … , Ye Zhang, Eric R. Gross
Published December 6, 2022
Citation Information: J Clin Invest. 2023;133(3):e163735. https://doi.org/10.1172/JCI163735.
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A human TRPV1 genetic variant within the channel gating domain regulates pain sensitivity in rodents

Abstract

Pain signals are relayed to the brain via a nociceptive system, and in rare cases, this nociceptive system contains genetic variants that can limit the pain response. Here, we questioned whether a human transient receptor potential vanilloid 1 (TRPV1) missense variant causes a resistance to noxious stimuli and, further, whether we could target this region with a cell-permeable peptide as a pain therapeutic. Initially using a computational approach, we identified a human K710N TRPV1 missense variant in an otherwise highly conserved region of mammalian TRPV1. After generating a TRPV1K710N-knockin mouse using CRISPR/Cas9, we discovered that the K710N variant reduced capsaicin-induced calcium influx in dorsal root ganglion neurons. The TRPV1K710N rodents also had less acute behavioral responses to noxious chemical stimuli and less hypersensitivity to nerve injury, while their response to noxious heat remained intact. Furthermore, blocking this K710 region in WT rodents using a cell-penetrating peptide limited acute behavioral responses to noxious stimuli and returned pain hypersensitivity induced by nerve injury to baseline levels. These findings identify K710 TRPV1 as a discrete site that is crucial for the control of nociception and provide insights into how to leverage rare genetic variants in humans to uncover fresh strategies for developing pain therapeutics.

Authors

Shufang He, Vanessa O. Zambelli, Pritam Sinharoy, Laura Brabenec, Yang Bian, Freeborn Rwere, Rafaela C.R. Hell, Beatriz Stein Neto, Barbara Hung, Xuan Yu, Meng Zhao, Zhaofei Luo, Chao Wu, Lijun Xu, Katrin J. Svensson, Stacy L. McAllister, Creed M. Stary, Nana-Maria Wagner, Ye Zhang, Eric R. Gross

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Figure 1

Identification of a human missense TRPV1 variant that reduces TRPV1-mediated calcium influx.

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Identification of a human missense TRPV1 variant that reduces TRPV1-medi...
(A) Venn diagram identifying 5 missense TRPV1 variants that intersect when examining the human TRPV1 missense variants from the gnomAD and avian genetic divergence from the human TRPV1 sequence. (B) Location of these 5 missense variants within the rat TRPV1 structure (PDB ID: 3J5P). Alignment of the mammalian and avian TRP domain (I696–S711 within the red dotted box; the unconserved amino acids 708 and 710 are shown in white). (C) 3D structure of WT TRPV1, K710N, and T708I based on the closed-state rat TRPV1 molecular model (PDB ID: 3J5P). Polar contacts are indicated in red or by blue dashed lines. (D–G) Calcium influx in response to 1 μM capsaicin (Cap) with (D) WT TRPV1, (E) K710N, (F) T708I, or (G) K710N/T708I TRPV1 mutations presented as a Fura-2AM ratio of 340:380 nm. (H) The AUC (total amount of calcium influx) and (I) percentage of maximal change in the Fura-2AM ratio for TRPV1 mutants relative to WT TRPV1 were calculated. n = 8 cells/group from 3 independent experiments. Data are expressed as the mean ± SEM. Significance was determined by 1-way ANOVA followed by Tukey’s post hoc test.