Abstract
Fibrotic disease is caused by the continuous deposition of extracellular matrix by persistently activated fibroblasts (also known as myofibroblasts), even after the resolution of the injury. Using fibroblasts from porcine aortic valves cultured on hydrogels that can be softened via exposure to ultraviolet light, here we show that increased extracellular stiffness activates the fibroblasts, and that cumulative tension on the nuclear membrane and increases in the activity of histone deacetylases transform transiently activated fibroblasts into myofibroblasts displaying condensed chromatin with genome-wide alterations. The condensed structure of the myofibroblasts is associated with cytoskeletal stability, as indicated by the inhibition of chromatin condensation and myofibroblast persistence after detachment of the nucleus from the cytoskeleton via the displacement of endogenous nesprins from the nuclear envelope. We also show that the chromatin structure of myofibroblasts from patients with aortic valve stenosis is more condensed than that of myofibroblasts from healthy donors. Our findings suggest that nuclear mechanosensing drives distinct chromatin signatures in persistently activated fibroblasts.
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Data availability
The authors declare that the main data supporting the results in this study are available within the paper and its Supplementary Information. The ATAC-seq data are available through the Gene Expression Omnibus under the accession code GSE167892. The raw and analysed datasets generated during the study are too large to be publicly shared, yet they are available from the corresponding authors on reasonable request.
Change history
28 May 2021
A Correction to this paper has been published: https://doi.org/10.1038/s41551-021-00748-3
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Acknowledgements
We acknowledge the support of the National Institutes of Health (grant no. R01 HL132353). C.J.W. was supported by the National Science Foundation (training grant no. IGERT 1144807), a National Institutes of Health Predoctoral Fellowship (grant no. F31HL142223) and a Department of Education GAANN Biomaterials Fellowship. A.R.K. was supported by the National Institutes of Health (grant no. R21 AR067469). C.C. was supported by a Human Frontiers Science Program fellowship (grant no. LT001449/2017‐L) and American Heart Association postdoctoral fellowship (grant no. 20POST3521111). J.C.G. was supported by a postdoctoral fellowship from the National Institutes of Health (grant no. T32 HL007822‐20). B.A.A. acknowledges funding from the National Institutes of Health (grant no. K99 HL148542) and the Burroughs Welcome Fund Postdoctoral Enrichment Program. L.A.L. was supported by the National Institutes of Health (grant nos RHL117138-05 and R01 GM29090). R.D.D. was supported by the National Institutes of Health (grant no. RO1 GM125871). We thank the BioFrontiers Institute Next-Gen Sequencing Core Facility, which performed the Illumina sequencing and library construction. The imaging work was performed at the BioFrontiers Institute Advanced Light Microscopy Core. Laser scanning confocal microscopy was performed on a Nikon A1R microscope supported by the NIST-CU Cooperative Agreement award number 70NANB15H226. Spinning disc confocal microscopy was performed on a Nikon Ti-E microscope supported by the BioFrontiers Institute and the Howard Hughes Medical Institute. We acknowledge the BioFrontiers Computing Core at the University of Colorado Boulder for providing high-performance computing resources (grant no. NIH 1S10OD012300) supported by BioFrontiers’ IT. C.J.W. thanks T. Ceccato, A. G. Rodriguez, M. Schroeder and D. Batan for their support and feedback on project ideas; and I. Marozas for help with plasmid acquisition and advice.
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C.J.W., L.A.L. and K.S.A. designed all of the experiments. M.A.A. and R.D.D. advised on the development and analysis of the ATAC-seq protocol. C.J.W. and D.R. performed the ATAC-seq experiment, and D.R. analysed the ATAC-seq data. C.J.W., C.C., A.R.K., J.C.G. and K.C. performed and analysed the cell culture experiments on hydrogels. C.J.W., C.C., D.R. and B.A.A. contributed to writing the paper. L.A.L. and K.S.A. edited the paper.
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Walker, C.J., Crocini, C., Ramirez, D. et al. Nuclear mechanosensing drives chromatin remodelling in persistently activated fibroblasts. Nat Biomed Eng 5, 1485–1499 (2021). https://doi.org/10.1038/s41551-021-00709-w
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DOI: https://doi.org/10.1038/s41551-021-00709-w
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