Skeletal dysplasia-causing TRPV4 mutations suppress the hypertrophic differentiation of human iPSC-derived chondrocytes
- Washington University in St. Louis, United States
- Regeneron Pharmaceuticals, United States
- 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).
Article and author information
Author details
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.
Reviewing Editor
- Di Chen, Chinese Academy of Sciences, China
Version history
- Received: June 10, 2021
- Preprint posted: June 15, 2021 (view preprint)
- Accepted: February 3, 2023
- Accepted Manuscript published: February 22, 2023 (version 1)
- Version of Record published: February 23, 2023 (version 2)
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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Skeletal dysplasia-causing TRPV4 mutations suppress the hypertrophic differentiation of human iPSC-derived chondrocytes
eLife 12:e71154.
https://doi.org/10.7554/eLife.71154
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