Cited by 530Cellular Mechanics and InteractionsInterstitial Lung Diseases and Idiopathic Pulmonary FibrosisProtein Kinase Regulation and GTPase Signaling

Matrix stiffness-induced myofibroblast differentiation is mediated by intrinsic mechanotransduction

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Huang, Xiangwei, Yang, Naiheng, Fiore, Vincent F., Barker, Thomas H., Sun, Yi, Morris, Stephan W., Ding, Qiang, Thannickal, Victor J. +1 more·2012·American Journal of Respiratory Cell and Molecular Biology

Abstract

The mechanical properties of the extracellular matrix have recently been shown to promote myofibroblast differentiation and lung fibrosis. Mechanisms by which matrix stiffness regulates myofibroblast differentiation are not fully understood. The goal of this study was to determine the intrinsic mechanisms of mechanotransduction in the regulation of matrix stiffness–induced myofibroblast differentiation. A well established polyacrylamide gel system with tunable substrate stiffness was used in this study. Megakaryoblastic leukemia factor-1 (MKL1) nuclear translocation was imaged by confocal immunofluorescent microscopy. The binding of MKL1 to the α-smooth muscle actin (α-SMA) gene promoter was quantified by quantitative chromatin immunoprecipitation assay. Normal human lung fibroblasts responded to matrix stiffening with changes in actin dynamics that favor filamentous actin polymerization. Actin polymerization resulted in nuclear translocation of MKL1, a serum response factor coactivator that plays a central role in regulating the expression of fibrotic genes, including α-SMA, a marker for myofibroblast differentiation. Mouse lung fibroblasts deficient in Mkl1 did not respond to matrix stiffening with increased α-SMA expression, whereas ectopic expression of human MKL1 cDNA restored the ability of Mkl1 null lung fibroblasts to express α-SMA. Furthermore, matrix stiffening promoted production and activation of the small GTPase RhoA, increased Rho kinase (ROCK) activity, and enhanced fibroblast contractility. Inhibition of RhoA/ROCK abrogated stiff matrix–induced actin cytoskeletal reorganization, MKL1 nuclear translocation, and myofibroblast differentiation. This study indicates that actin cytoskeletal remodeling–mediated activation of MKL1 transduces mechanical stimuli from the extracellular matrix to a fibrogenic program that promotes myofibroblast differentiation, suggesting an intrinsic mechanotransduction mechanism.

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  1. Desmouliere A, Chaponnier C, Gabbiani G. Tissue repair, contraction, and the myofibroblast. Wound Repair Regen 2005;13:7-12.
  2. Hooley R, Yu CY, Symons M, Barber DL. G alpha 13 stimulates na1- h1 exchange through distinct cdc42-dependent and rhoa-dependent pathways. J Biol Chem 1996;271:6152-6158.
  3. Danen EH, Sonneveld P, Brakebusch C, Fassler R, Sonnenberg A. The fibronectin-binding integrins alpha5beta1 and alphavbeta3 differen- tially modulate rhoa-gtp loading, organization of cell matrix adhe- sions, and fibronectin fibrillogenesis. J Cell Biol 2002;159:1071-1086.

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Authors
Huang, X., Yang, N., Fiore, V. F., Barker, T. H., Sun, Y., Morris, S. W., Ding, Q., Thannickal, V. J., & Zhou, Y.
Affiliations
  • University of Alabama at Birmingham (US)
  • Georgia Institute of Technology (US)
  • St. Jude Children's Research Hospital (US)
Corresponding author
Zhou, Yong
Venue
American Journal of Respiratory Cell and Molecular Biology
Published
2012-09-01
Volume
47
Issue
3
Pages
340-348
ISSN
  • 1044-1549 (ISSN-L)
  • 1535-4989
OpenAlex
W2172237567
Cited by
530
References
65
Citations by year
2026: 39 · 2025: 35 · 2024: 44 · 2023: 45 · 2022: 35 · 2021: 44 · 2020: 41 · 2019: 34 · 2018: 47 · 2017: 36 · 2016: 22 · 2015: 39 · 2014: 26 · 2013: 32 · 2012: 11
FWCI
16.35
Type
journal article
Language
en
Open access
Locations
  • American Journal of Respiratory Cell and Molecular Biology (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Science Foundation
  • American Heart Association
  • St. Jude Children's Research Hospital
  • American Lebanese Syrian Associated Charities
  • National Institutes of Health
  • National Heart, Lung, and Blood Institute (R21HL097215, HL107181, HL 067967, P50HL107181, R01HL067967, R01HL085324, HL097215)
  • National Cancer Institute (R01CA087064, CA87064)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Interstitial Lung Diseases and Idiopathic Pulmonary Fibrosis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
Keywords
extracellular matrix stiffness, myofibroblast differentiation, mechanotransduction, MKL1, RhoA/ROCK signaling, actin cytoskeleton remodeling, alpha-smooth muscle actin, pulmonary fibrosis, lung fibroblasts
MeSH terms
Animals, Base Sequence, Cell Differentiation, Cells, Cultured, DNA-Binding Proteins, Electrophoresis, Polyacrylamide Gel, Extracellular Matrix, Humans, Phosphorylation, Oncogene Proteins, Fusion, Trans-Activators, Extracellular Matrix Proteins, DNA Primers, Mice, Knockout, Mechanotransduction, Cellular, Mice, Myofibroblasts

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