YAP/TAZ are essential for TGF-β2–mediated conjunctival fibrosis

Akiko Futakuchi, Toshihiro Inoue, Fan Yan Wei, Miyuki Inoue-Mochita, Tomokazu Fujimoto, Kazuhito Tomizawa, Hidenobu Tanihara

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

PURPOSE. To investigate the roles of Yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ), the major effector molecules of the Hippo pathway, in TGF-β2–mediated conjunctival fibrosis. METHODS. Primary human conjunctival fibroblasts were treated with TGF-β2. The expression of YAP/TAZ was examined by Western blot analyses and immunocytochemistry. The expression of fibrotic proteins and genes were evaluated by Western blot analyses and quantitative real-time PCR, respectively. The effects of YAP/TAZ on fibrotic changes were examined by knockdown experiments and the YAP/TAZ inhibitor, verteporfin. RESULTS. TGF-β2 stabilized YAP/TAZ and subsequently activated Smad2/3, which led to the transcription of fibrotic genes in human primary conjunctival fibroblasts. These fibrotic genes were differently regulated by YAP/TAZ. Notably, a-smooth muscle actin, fibronectin, collagen I, and collagen IV were primarily regulated by YAP. In contrast, CCN family proteins (CTGF and CYR61) depended on both YAP and TAZ. Mechanistically, YAP/TAZ were located in close proximity to Smad2/3, and in particular, YAP was required for TGF-β2-mediated phosphorylation and the nuclear translocation of Smad2/3. Furthermore, a YAP/TAZ inhibitor markedly suppressed TGF-β2-mediated fibrotic changes in conjunctival fibroblasts. CONCLUSIONS. YAP/TAZ acted as a molecular hub of TGF-β2 signaling in a cellular model of conjunctival fibrosis. Moreover, verteporfin, a YAP/TAZ inhibitor exerted potent antifibrosis effects by suppressing TGF-β2-YAP/TAZ-Smad signaling. Our study highlights YAP/TAZ as essential regulators of conjunctival fibrosis and shows that inhibition of YAP/TAZ might potentially improve the outcomes of glaucoma filtration surgery.

Original languageEnglish
Pages (from-to)3069-3078
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume59
Issue number7
DOIs
Publication statusPublished - Jun 1 2018
Externally publishedYes

Fingerprint

Fibrosis
Proteins
Fibroblasts
CCN Intercellular Signaling Proteins
Collagen
Western Blotting
Smad Proteins
Filtering Surgery
Fibronectins
Glaucoma
Genes
Smooth Muscle
Actins
Real-Time Polymerase Chain Reaction
Immunohistochemistry
Phosphorylation

Keywords

  • Fibrosis
  • Glaucoma
  • TGF-β
  • The Hippo pathway

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Futakuchi, A., Inoue, T., Wei, F. Y., Inoue-Mochita, M., Fujimoto, T., Tomizawa, K., & Tanihara, H. (2018). YAP/TAZ are essential for TGF-β2–mediated conjunctival fibrosis. Investigative Ophthalmology and Visual Science, 59(7), 3069-3078. https://doi.org/10.1167/iovs.18-24258

YAP/TAZ are essential for TGF-β2–mediated conjunctival fibrosis. / Futakuchi, Akiko; Inoue, Toshihiro; Wei, Fan Yan; Inoue-Mochita, Miyuki; Fujimoto, Tomokazu; Tomizawa, Kazuhito; Tanihara, Hidenobu.

In: Investigative Ophthalmology and Visual Science, Vol. 59, No. 7, 01.06.2018, p. 3069-3078.

Research output: Contribution to journalArticle

Futakuchi, A, Inoue, T, Wei, FY, Inoue-Mochita, M, Fujimoto, T, Tomizawa, K & Tanihara, H 2018, 'YAP/TAZ are essential for TGF-β2–mediated conjunctival fibrosis', Investigative Ophthalmology and Visual Science, vol. 59, no. 7, pp. 3069-3078. https://doi.org/10.1167/iovs.18-24258
Futakuchi A, Inoue T, Wei FY, Inoue-Mochita M, Fujimoto T, Tomizawa K et al. YAP/TAZ are essential for TGF-β2–mediated conjunctival fibrosis. Investigative Ophthalmology and Visual Science. 2018 Jun 1;59(7):3069-3078. https://doi.org/10.1167/iovs.18-24258
Futakuchi, Akiko ; Inoue, Toshihiro ; Wei, Fan Yan ; Inoue-Mochita, Miyuki ; Fujimoto, Tomokazu ; Tomizawa, Kazuhito ; Tanihara, Hidenobu. / YAP/TAZ are essential for TGF-β2–mediated conjunctival fibrosis. In: Investigative Ophthalmology and Visual Science. 2018 ; Vol. 59, No. 7. pp. 3069-3078.
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abstract = "PURPOSE. To investigate the roles of Yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ), the major effector molecules of the Hippo pathway, in TGF-β2–mediated conjunctival fibrosis. METHODS. Primary human conjunctival fibroblasts were treated with TGF-β2. The expression of YAP/TAZ was examined by Western blot analyses and immunocytochemistry. The expression of fibrotic proteins and genes were evaluated by Western blot analyses and quantitative real-time PCR, respectively. The effects of YAP/TAZ on fibrotic changes were examined by knockdown experiments and the YAP/TAZ inhibitor, verteporfin. RESULTS. TGF-β2 stabilized YAP/TAZ and subsequently activated Smad2/3, which led to the transcription of fibrotic genes in human primary conjunctival fibroblasts. These fibrotic genes were differently regulated by YAP/TAZ. Notably, a-smooth muscle actin, fibronectin, collagen I, and collagen IV were primarily regulated by YAP. In contrast, CCN family proteins (CTGF and CYR61) depended on both YAP and TAZ. Mechanistically, YAP/TAZ were located in close proximity to Smad2/3, and in particular, YAP was required for TGF-β2-mediated phosphorylation and the nuclear translocation of Smad2/3. Furthermore, a YAP/TAZ inhibitor markedly suppressed TGF-β2-mediated fibrotic changes in conjunctival fibroblasts. CONCLUSIONS. YAP/TAZ acted as a molecular hub of TGF-β2 signaling in a cellular model of conjunctival fibrosis. Moreover, verteporfin, a YAP/TAZ inhibitor exerted potent antifibrosis effects by suppressing TGF-β2-YAP/TAZ-Smad signaling. Our study highlights YAP/TAZ as essential regulators of conjunctival fibrosis and shows that inhibition of YAP/TAZ might potentially improve the outcomes of glaucoma filtration surgery.",
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T1 - YAP/TAZ are essential for TGF-β2–mediated conjunctival fibrosis

AU - Futakuchi, Akiko

AU - Inoue, Toshihiro

AU - Wei, Fan Yan

AU - Inoue-Mochita, Miyuki

AU - Fujimoto, Tomokazu

AU - Tomizawa, Kazuhito

AU - Tanihara, Hidenobu

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N2 - PURPOSE. To investigate the roles of Yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ), the major effector molecules of the Hippo pathway, in TGF-β2–mediated conjunctival fibrosis. METHODS. Primary human conjunctival fibroblasts were treated with TGF-β2. The expression of YAP/TAZ was examined by Western blot analyses and immunocytochemistry. The expression of fibrotic proteins and genes were evaluated by Western blot analyses and quantitative real-time PCR, respectively. The effects of YAP/TAZ on fibrotic changes were examined by knockdown experiments and the YAP/TAZ inhibitor, verteporfin. RESULTS. TGF-β2 stabilized YAP/TAZ and subsequently activated Smad2/3, which led to the transcription of fibrotic genes in human primary conjunctival fibroblasts. These fibrotic genes were differently regulated by YAP/TAZ. Notably, a-smooth muscle actin, fibronectin, collagen I, and collagen IV were primarily regulated by YAP. In contrast, CCN family proteins (CTGF and CYR61) depended on both YAP and TAZ. Mechanistically, YAP/TAZ were located in close proximity to Smad2/3, and in particular, YAP was required for TGF-β2-mediated phosphorylation and the nuclear translocation of Smad2/3. Furthermore, a YAP/TAZ inhibitor markedly suppressed TGF-β2-mediated fibrotic changes in conjunctival fibroblasts. CONCLUSIONS. YAP/TAZ acted as a molecular hub of TGF-β2 signaling in a cellular model of conjunctival fibrosis. Moreover, verteporfin, a YAP/TAZ inhibitor exerted potent antifibrosis effects by suppressing TGF-β2-YAP/TAZ-Smad signaling. Our study highlights YAP/TAZ as essential regulators of conjunctival fibrosis and shows that inhibition of YAP/TAZ might potentially improve the outcomes of glaucoma filtration surgery.

AB - PURPOSE. To investigate the roles of Yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ), the major effector molecules of the Hippo pathway, in TGF-β2–mediated conjunctival fibrosis. METHODS. Primary human conjunctival fibroblasts were treated with TGF-β2. The expression of YAP/TAZ was examined by Western blot analyses and immunocytochemistry. The expression of fibrotic proteins and genes were evaluated by Western blot analyses and quantitative real-time PCR, respectively. The effects of YAP/TAZ on fibrotic changes were examined by knockdown experiments and the YAP/TAZ inhibitor, verteporfin. RESULTS. TGF-β2 stabilized YAP/TAZ and subsequently activated Smad2/3, which led to the transcription of fibrotic genes in human primary conjunctival fibroblasts. These fibrotic genes were differently regulated by YAP/TAZ. Notably, a-smooth muscle actin, fibronectin, collagen I, and collagen IV were primarily regulated by YAP. In contrast, CCN family proteins (CTGF and CYR61) depended on both YAP and TAZ. Mechanistically, YAP/TAZ were located in close proximity to Smad2/3, and in particular, YAP was required for TGF-β2-mediated phosphorylation and the nuclear translocation of Smad2/3. Furthermore, a YAP/TAZ inhibitor markedly suppressed TGF-β2-mediated fibrotic changes in conjunctival fibroblasts. CONCLUSIONS. YAP/TAZ acted as a molecular hub of TGF-β2 signaling in a cellular model of conjunctival fibrosis. Moreover, verteporfin, a YAP/TAZ inhibitor exerted potent antifibrosis effects by suppressing TGF-β2-YAP/TAZ-Smad signaling. Our study highlights YAP/TAZ as essential regulators of conjunctival fibrosis and shows that inhibition of YAP/TAZ might potentially improve the outcomes of glaucoma filtration surgery.

KW - Fibrosis

KW - Glaucoma

KW - TGF-β

KW - The Hippo pathway

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