Journal article
TRPV4-mediated calcium signaling in mesenchymal stem cells regulates aligned collagen matrix formation and vinculin tension
Proceedings of the National Academy of Sciences - PNAS, Vol.116(6), pp.1992-1997
02/05/2019
DOI: 10.1073/pnas.1811095116
PMCID: PMC6369759
PMID: 30674675
Abstract
Microarchitectural cues drive aligned fibrillar collagen deposition in vivo and in biomaterial scaffolds, but the cell-signaling events that underlie this process are not well understood. Utilizing a multicellular patterning model system that allows for observation of intracellular signaling events during collagen matrix assembly, we investigated the role of calcium (Ca
) signaling in human mesenchymal stem cells (MSCs) during this process. We observed spontaneous Ca
oscillations in MSCs during fibrillar collagen assembly, and hypothesized that the transient receptor potential vanilloid 4 (TRPV4) ion channel, a mechanosensitive Ca
-permeable channel, may regulate this signaling. Inhibition of TRPV4 nearly abolished Ca
signaling at initial stages of collagen matrix assembly, while at later times had reduced but significant effects. Importantly, blocking TRPV4 activity dramatically reduced aligned collagen fibril assembly; conversely, activating TRPV4 accelerated aligned collagen formation. TRPV4-dependent Ca
oscillations were found to be independent of pattern shape or subpattern cell location, suggesting this signaling mechanism is necessary for aligned collagen formation but not sufficient in the absence of physical (microarchitectural) cues that force multicellular alignment. As cell-generated mechanical forces are known to be critical to the matrix assembly process, we examined the role of TRPV4-mediated Ca
signaling in force generated across the load-bearing focal adhesion protein vinculin within MSCs using an FRET-based tension sensor. Inhibiting TRPV4 decreased tensile force across vinculin, whereas TRPV4 activation caused a dynamic unloading and reloading of vinculin. Together, these findings suggest TRPV4 activity regulates forces at cell-matrix adhesions and is critical to aligned collagen matrix assembly by MSCs.
Details
- Title: Subtitle
- TRPV4-mediated calcium signaling in mesenchymal stem cells regulates aligned collagen matrix formation and vinculin tension
- Creators
- Christopher L Gilchrist - Duke UniversityHolly A Leddy - Duke UniversityLaurel Kaye - Duke UniversityNatasha D Case - Saint Louis UniversityKatheryn E Rothenberg - Duke UniversityDianne Little - Purdue University West LafayetteWolfgang Liedtke - Duke University HospitalBrenton D Hoffman - Duke UniversityFarshid Guilak - Washington University in St. Louis
- Resource Type
- Journal article
- Publication Details
- Proceedings of the National Academy of Sciences - PNAS, Vol.116(6), pp.1992-1997
- DOI
- 10.1073/pnas.1811095116
- PMID
- 30674675
- PMCID
- PMC6369759
- ISSN
- 0027-8424
- eISSN
- 1091-6490
- Grant note
- P30 AR057235 / NIAMS NIH HHS R01 AR048182 / NIAMS NIH HHS R01 AG015768 / NIA NIH HHS K08 AR059784 / NIAMS NIH HHS F32 AR065888 / NIAMS NIH HHS P01 AR050245 / NIAMS NIH HHS R01 AR048852 / NIAMS NIH HHS R01 AG046927 / NIA NIH HHS P30 AR073752 / NIAMS NIH HHS R03 AR065764 / NIAMS NIH HHS
- Language
- English
- Date published
- 02/05/2019
- Academic Unit
- Biology
- Record Identifier
- 9984696721202771
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