RNAi screening identifies a mechanosensitive ROCK-JAK2- STAT3 network central to myofibroblast activation

Raymond S. Oh, Andrew J. Haak, Karry M.J. Smith, Giovanni Ligresti, Kyoung Moo Choi, Tiao Xie, Shaohua Wang, Paula R. Walters, Michael A. Thompson, Michelle R. Freeman, Logan J. Manlove, Vivian M. Chu, Carol Feghali-Bostwick, Anja Roden, Jürgen Schymeinsky, Christina M Pabelick, Y.s. Prakash, Robert Vassallo, Daniel J Tschumperlin

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Myofibroblasts play key roles in wound healing and pathological fibrosis. Here, we used an RNAi screen to characterize myofibroblast regulatory genes, using a high-content imaging approach to quantify a-smooth muscle actin stress fibers in cultured human fibroblasts. Screen hits were validated on physiological compliance hydrogels, and selected hits tested in primary fibroblasts from patients with idiopathic pulmonary fibrosis. Our RNAi screen led to the identification of STAT3 as an essential mediator of myofibroblast activation and function. Strikingly, we found that STAT3 phosphorylation, while responsive to exogenous ligands on both soft and stiff matrices, is innately active on a stiff matrix in a ligand/receptor-independent, but ROCK- and JAK2-dependent fashion. These results demonstrate how a cytokine-inducible signal can become persistently activated by pathological matrix stiffening. Consistent with a pivotal role for this pathway in driving persistent fibrosis, a STAT3 inhibitor attenuated murine pulmonary fibrosis when administered in a therapeutic fashion after bleomycin injury. Our results identify novel genes essential for the myofibroblast phenotype, and point to STAT3 as an important target in pulmonary fibrosis and other fibrotic diseases.

Original languageEnglish (US)
Article numberjcs209932
JournalJournal of Cell Science
Volume131
Issue number10
DOIs
StatePublished - May 15 2018

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Keywords

  • Fibroblast
  • Fibrosis
  • SiRNA
  • Stiffness
  • TGF-β

ASJC Scopus subject areas

  • Cell Biology

Cite this

Oh, R. S., Haak, A. J., Smith, K. M. J., Ligresti, G., Choi, K. M., Xie, T., Wang, S., Walters, P. R., Thompson, M. A., Freeman, M. R., Manlove, L. J., Chu, V. M., Feghali-Bostwick, C., Roden, A., Schymeinsky, J., Pabelick, C. M., Prakash, Y. S., Vassallo, R., & Tschumperlin, D. J. (2018). RNAi screening identifies a mechanosensitive ROCK-JAK2- STAT3 network central to myofibroblast activation. Journal of Cell Science, 131(10), [jcs209932]. https://doi.org/10.1242/jcs.209932