Skeletal dysplasia-causing TRPV4 mutations suppress the hypertrophic differentiation of human iPSC-derived chondrocytes

  1. Amanda R Dicks
  2. Grigory I Maksaev
  3. Zainab Harissa
  4. Alireza Savadipour
  5. Ruhang Tang
  6. Nancy Steward
  7. Wolfgang Liedtke
  8. Colin G Nichols
  9. Chia-Lung Wu
  10. Farshid Guilak  Is a corresponding author
  1. Washington University in St. Louis, United States
  2. Regeneron Pharmaceuticals, United States
  3. University of Rochester, United States

Abstract

Mutations in the TRPV4 ion channel can lead to a range of skeletal dysplasias. However, the mechanisms by which TRPV4 mutations lead to distinct disease severity remain unknown. Here, we use CRISPR-Cas9-edited human induced pluripotent stem cells (hiPSCs) harboring either the mild V620I or lethal T89I mutations to elucidate the differential effects on channel function and chondrogenic differentiation. We found that hiPSC-derived chondrocytes with the V620I mutation exhibited increased basal currents through TRPV4. However, both mutations showed more rapid calcium signaling with a reduced overall magnitude in response to TRPV4 agonist GSK1016790A compared to wildtype. There were no differences in overall cartilaginous matrix production, but the V620I mutation resulted in reduced mechanical properties of cartilage matrix later in chondrogenesis. mRNA sequencing revealed that both mutations upregulated several anterior HOX genes and downregulated antioxidant genes CAT and GSTA1 throughout chondrogenesis. BMP4 treatment upregulated several essential hypertrophic genes in WT chondrocytes; however, this hypertrophic maturation response was inhibited in mutant chondrocytes. These results indicate that the TRPV4 mutations alter BMP signaling in chondrocytes and prevent proper chondrocyte hypertrophy, as a potential mechanism for dysfunctional skeletal development. Our findings provide potential therapeutic targets for developing treatments for TRPV4-mediated skeletal dysplasias.

Data availability

All RNAseq data files generated and reported in this study are available on GEO (accession number GSE225446, https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE225446).

The following data sets were generated

Article and author information

Author details

  1. Amanda R Dicks

    Department of Biomedical Engineering, Washington University in St. Louis, St Louis, United States
    Competing interests
    No competing interests declared.
  2. Grigory I Maksaev

    Department of Cell Biology and Physiology, Washington University in St. Louis, St Louis, United States
    Competing interests
    No competing interests declared.
  3. Zainab Harissa

    Department of Biomedical Engineering, Washington University in St. Louis, St Louis, United States
    Competing interests
    No competing interests declared.
  4. Alireza Savadipour

    Department of Orthopedic Surgery, Washington University in St. Louis, St Louis, United States
    Competing interests
    No competing interests declared.
  5. Ruhang Tang

    Department of Orthopedic Surgery, Washington University in St. Louis, St Louis, United States
    Competing interests
    No competing interests declared.
  6. Nancy Steward

    Department of Orthopedic Surgery, Washington University in St. Louis, St Louis, United States
    Competing interests
    No competing interests declared.
  7. Wolfgang Liedtke

    Regeneron Pharmaceuticals, Tarrytown, United States
    Competing interests
    Wolfgang Liedtke, Patents on TRPV4 inhibitors have been licensed to TRPblue. Dr. Liedtke is an employee of Regeneron Pharmaceuticals.(US Patents 9,701,675; 10,329,265; and 11,014,896)..
  8. Colin G Nichols

    Department of Cell Biology and Physiology, Washington University in St. Louis, St Louis, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4929-2134
  9. Chia-Lung Wu

    Department of Orthopaedics and Rehabilitation, University of Rochester, Rochester, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9598-7036
  10. Farshid Guilak

    Department of Orthopedic Surgery, Washington University in St. Louis, St Louis, United States
    For correspondence
    guilak@wustl.edu
    Competing interests
    Farshid Guilak, Patents on TRPV4 inhibitors licensed to TRPblue Inc. (US Patents 9,701,675; 10,329,265; and 11,014,896)..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7380-0330

Funding

National Institutes of Health (AG15768)

  • Farshid Guilak

National Institutes of Health (AG46927)

  • Farshid Guilak

National Institutes of Health (ar072999)

  • Farshid Guilak

National Institutes of Health (AR075899)

  • Chia-Lung Wu

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

Copyright

© 2023, Dicks 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.

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  1. Amanda R Dicks
  2. Grigory I Maksaev
  3. Zainab Harissa
  4. Alireza Savadipour
  5. Ruhang Tang
  6. Nancy Steward
  7. Wolfgang Liedtke
  8. Colin G Nichols
  9. Chia-Lung Wu
  10. Farshid Guilak
(2023)
Skeletal dysplasia-causing TRPV4 mutations suppress the hypertrophic differentiation of human iPSC-derived chondrocytes
eLife 12:e71154.
https://doi.org/10.7554/eLife.71154

Share this article

https://doi.org/10.7554/eLife.71154

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