Therapeutic inhibition of keratinocyte TRPV3 sensory channel by local anesthetic dyclonine

  1. Qiang Liu
  2. Jin Wang
  3. Xin Wei
  4. Juan Hu
  5. Conghui Ping
  6. Yue Gao
  7. Chang Xie
  8. Peiyu Wang
  9. Peng Cao
  10. Zhengyu Cao
  11. Ye Yu
  12. Dongdong Li
  13. Jing Yao  Is a corresponding author
  1. Wuhan University, China
  2. China Pharmaceutical University, China
  3. Nanjing University of Chinese Medicine, China
  4. CNRS UMR8246, INSERM U1130, UPMC UM119, France

Abstract

The multimodal sensory channel transient receptor potential vanilloid-3 (TRPV3) is expressed in epidermal keratinocytes and implicated in chronic pruritus, allergy, and inflammation-related skin disorders. Gain-of-function mutations of TRPV3 cause hair growth disorders in mice and Olmsted Syndrome in human. We here report that mouse and human TRPV3 channel is targeted by the clinical medication dyclonine that exerts a potent inhibitory effect. Accordingly, dyclonine rescued cell death caused by gain-of-function TRPV3 mutations and suppressed pruritus symptoms in vivo in mouse model. At the single-channel level, dyclonine inhibited TRPV3 open probability but not the unitary conductance. By molecular simulations and mutagenesis, we further uncovered key residues in TRPV3 pore region that could toggle the inhibitory efficiency of dyclonine. The functional and mechanistic insights obtained on dyclonine-TRPV3 interaction will help to conceive updated therapeutics for skin inflammation.

Data availability

All the data for Therapeutic inhibition of keratinocyte TRPV3 sensory channel by local anesthetic dyclonine have been deposited in Dyrad with DOI https://doi.org/10.5061/dryad.7d7wm37sq.

The following data sets were generated

Article and author information

Author details

  1. Qiang Liu

    Department of Cell Biology, Wuhan University, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Jin Wang

    School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Xin Wei

    Department of Cell Biology, Wuhan University, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Juan Hu

    Department of Cell Biology, Wuhan University, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Conghui Ping

    Department of Cell Biology, Wuhan University, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  6. Yue Gao

    Department of Cell Biology, Wuhan University, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  7. Chang Xie

    Department of Cell Biology, Wuhan University, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  8. Peiyu Wang

    Department of Cell Biology, Wuhan University, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  9. Peng Cao

    Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, China
    Competing interests
    The authors declare that no competing interests exist.
  10. Zhengyu Cao

    School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, China
    Competing interests
    The authors declare that no competing interests exist.
  11. Ye Yu

    School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, China
    Competing interests
    The authors declare that no competing interests exist.
  12. Dongdong Li

    Neuroscience Paris Seine, CNRS UMR8246, INSERM U1130, UPMC UM119, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  13. Jing Yao

    Department of Cell Biology, Wuhan University, Wuhan, China
    For correspondence
    jyao@whu.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1844-3988

Funding

National Natural Science Foundation of China (31830031)

  • Jing Yao

National Natural Science Foundation of China (31929003)

  • Jing Yao

National Natural Science Foundation of China (31871174)

  • Jing Yao

National Natural Science Foundation of China (31671209)

  • Jing Yao

National Natural Science Foundation of China (31601864)

  • Jing Yao

Natural Science Foundation of Hubei Province (2017CFA063)

  • Jing Yao

Natural Science Foundation of Hubei Province (2018CFA016)

  • Jing Yao

Natural Science Foundation of Jiangsu Province (BK20202002)

  • Ye Yu

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All mice were housed in the specific pathogen-free animal facility at Wuhan University and all animal experiments were in accordance with protocols were adhered to the Chinese National Laboratory Animal-Guideline for Ethical Review of Animal Welfare and approved by the Institutional Animal Care and Use Committee of Wuhan University (NO. WDSKY0201804). The mice were euthanatized with CO2 followed by various studies.

Reviewing Editor

  1. Kenton J Swartz, National Institute of Neurological Disorders and Stroke, National Institutes of Health, United States

Version history

  1. Received: March 5, 2021
  2. Accepted: April 19, 2021
  3. Accepted Manuscript published: April 20, 2021 (version 1)
  4. Version of Record published: May 11, 2021 (version 2)
  5. Version of Record updated: May 13, 2021 (version 3)

Copyright

© 2021, Liu et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,988
    Page views
  • 326
    Downloads
  • 12
    Citations

Article citation count generated by polling the highest count across the following sources: PubMed Central, Crossref, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Qiang Liu
  2. Jin Wang
  3. Xin Wei
  4. Juan Hu
  5. Conghui Ping
  6. Yue Gao
  7. Chang Xie
  8. Peiyu Wang
  9. Peng Cao
  10. Zhengyu Cao
  11. Ye Yu
  12. Dongdong Li
  13. Jing Yao
(2021)
Therapeutic inhibition of keratinocyte TRPV3 sensory channel by local anesthetic dyclonine
eLife 10:e68128.
https://doi.org/10.7554/eLife.68128

Share this article

https://doi.org/10.7554/eLife.68128

Further reading

    1. Biochemistry and Chemical Biology
    2. Cell Biology
    Kristian Davidsen, Jonathan S Marvin ... Lucas B Sullivan
    Research Article

    Intracellular levels of the amino acid aspartate are responsive to changes in metabolism in mammalian cells and can correspondingly alter cell function, highlighting the need for robust tools to measure aspartate abundance. However, comprehensive understanding of aspartate metabolism has been limited by the throughput, cost, and static nature of the mass spectrometry (MS)-based measurements that are typically employed to measure aspartate levels. To address these issues, we have developed a green fluorescent protein (GFP)-based sensor of aspartate (jAspSnFR3), where the fluorescence intensity corresponds to aspartate concentration. As a purified protein, the sensor has a 20-fold increase in fluorescence upon aspartate saturation, with dose-dependent fluorescence changes covering a physiologically relevant aspartate concentration range and no significant off target binding. Expressed in mammalian cell lines, sensor intensity correlated with aspartate levels measured by MS and could resolve temporal changes in intracellular aspartate from genetic, pharmacological, and nutritional manipulations. These data demonstrate the utility of jAspSnFR3 and highlight the opportunities it provides for temporally resolved and high-throughput applications of variables that affect aspartate levels.

    1. Biochemistry and Chemical Biology
    Chi-Ning Chuang, Hou-Cheng Liu ... Ting-Fang Wang
    Research Article

    Serine(S)/threonine(T)-glutamine(Q) cluster domains (SCDs), polyglutamine (polyQ) tracts and polyglutamine/asparagine (polyQ/N) tracts are Q-rich motifs found in many proteins. SCDs often are intrinsically disordered regions that mediate protein phosphorylation and protein-protein interactions. PolyQ and polyQ/N tracts are structurally flexible sequences that trigger protein aggregation. We report that due to their high percentages of STQ or STQN amino acid content, four SCDs and three prion-causing Q/N-rich motifs of yeast proteins possess autonomous protein expression-enhancing activities. Since these Q-rich motifs can endow proteins with structural and functional plasticity, we suggest that they represent useful toolkits for evolutionary novelty. Comparative Gene Ontology (GO) analyses of the near-complete proteomes of 26 representative model eukaryotes reveal that Q-rich motifs prevail in proteins involved in specialized biological processes, including Saccharomyces cerevisiae RNA-mediated transposition and pseudohyphal growth, Candida albicans filamentous growth, ciliate peptidyl-glutamic acid modification and microtubule-based movement, Tetrahymena thermophila xylan catabolism and meiosis, Dictyostelium discoideum development and sexual cycles, Plasmodium falciparum infection, and the nervous systems of Drosophila melanogaster, Mus musculus and Homo sapiens. We also show that Q-rich-motif proteins are expanded massively in 10 ciliates with reassigned TAAQ and TAGQ codons. Notably, the usage frequency of CAGQ is much lower in ciliates with reassigned TAAQ and TAGQ codons than in organisms with expanded and unstable Q runs (e.g. D. melanogaster and H. sapiens), indicating that the use of noncanonical stop codons in ciliates may have coevolved with codon usage biases to avoid triplet repeat disorders mediated by CAG/GTC replication slippage.