A human TRPV1 genetic variant within the channel gating domain regulates pain sensitivity in rodents
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
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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Research Article Neuroscience Vascular biology

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 9

V1-cal rescues nocifensive behavior in WT TRPV1 mice after SNI.

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V1-cal rescues nocifensive behavior in WT TRPV1 mice after SNI. (A) Expe...
(A) Experimental protocol for SNI model with peptide treatment. Rodents underwent SNI and were assessed for 2 weeks (injury phase). After the injury phase, osmotic pumps were implanted to deliver V1-cal or TAT47–57 for 2 weeks (rescue phase). (B) Thermal latency for sham-treated (dashed line) and SNI (solid line) mice. SNI mice were treated with V1-cal (green) or TAT47–57 (purple) during the rescue phase. (C) Threshold of withdrawal from mechanical stimuli for sham-treated (dashed line) and for SNI (solid line) mice. SNI mice were treated with V1-cal (green) or TAT47–57 (purple) during the rescue phase. (D) Percentage of hypersensitivity relative to the uninjured paw for sham-treated (n = 8) and SNI (n = 17) mice treated with V1-cal (n = 9) or TAT47–57 (n = 8) during the injury and rescue phases. Data are expressed as the mean ± SEM. (E) Duration of nociceptive behavior (paw licking/flinching) following exposure to acetone for sham-treated (dashed line, n = 8) and SNI (solid line, n = 16) mice. SNI mice were treated with V1-cal (green, n = 8) or TAT47–57 (purple, n = 8) during the rescue phase. *P < 0.05 and **P < 0.01, SNI plus TAT47–57 versus sham; #P < 0.05 and ##P < 0.01, SNI plus TAT47–57 versus SNI plus V1-cal. Significance was determined by 2-way RM ANOVA analysis with a mixed-effects model followed by Bonferroni’s post hoc test (B, C, and E) and unpaired, 2-tailed t test for the injury phase and 1-way ANOVA followed by Tukey’s post hoc test for the rescue phase (D).