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Inhibition of Connective Tissue Growth Factor Expression in Adult Retinal Pigment Epithelial-19 Cells by Blocking
Yoko Murakami1, Toshiyasu Imaizumi1, Kouhei Hashizume1
1Department of Ophthalmology, School of Medicine, Iwate Medical University, 2-1-1 Idaitori, Yahaba-Cho, Shiwa-gun, Iwate, Japan.
Abstract:
To investigate the effect of yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) on connective tissue growth factor (CTGF) expression in adult retinal pigment epithelial (ARPE)-19 cells. We also studied the inhibitory effect of K-975, a new pan-transcriptional enhanced associate domain (TEAD) inhibitor, and luteolin, a plant-derived flavonoid on CTGF expression. ARPE-19 cells were transfected with either YAP or TAZ overexpression plasmid or treated with transforming growth factor (TGF)-β2. The cells were cultured either with or without K-975 or luteolin. The expression of YAP, TAZ, and CTGF was examined using real-time PCR. ARPE-19 cells overexpressing YAP or TAZ exhibited significantly increased CTGF expression. This increase was attenuated by K-975 or luteolin alone. TGF-β2 treatment significantly raised the expression of not just YAP and TAZ, but also CTGF in ARPE-19 cells. TGF-β2 treatment-enhanced CTGF expression was considerably lowered by the addition of K-975 or luteolin. Overexpression of YAP or TAZ and treatment with TGF-β2 led to an increase in the expression of CTGF in ARPE-19 cells. These increases were attenuated by treatment with K-975 and luteolin. These findings suggest that YAP and TAZ may be related to the expression of CTGF in ARPE-19 cells and that K-975 and luteolin can be explored as potential therapeutic agents for preventing CTGF production in vitreoretinal fibrosis.
Insights
Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) increase connective tissue growth factor (CTGF) in retinal cells. K-975 and luteolin inhibit this CTGF production, suggesting therapeutic potential for vitreoretinal fibrosis.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- Connective tissue growth factor (CTGF) plays a role in vitreoretinal fibrosis.
- The Hippo pathway, involving yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ), regulates cellular processes.
- Understanding the regulation of CTGF is crucial for developing treatments for fibrotic eye diseases.
Purpose of the Study:
- To investigate the role of YAP and TAZ in regulating CTGF expression in adult retinal pigment epithelial (ARPE)-19 cells.
- To evaluate the inhibitory effects of a TEAD inhibitor (K-975) and luteolin on CTGF expression.
- To explore potential therapeutic strategies for vitreoretinal fibrosis.
Main Methods:
- ARPE-19 cells were transfected with YAP or TAZ overexpression plasmids or treated with transforming growth factor-beta2 (TGF-β2).
- Cells were treated with or without K-975 or luteolin.
- Expression levels of YAP, TAZ, and CTGF were quantified using real-time PCR.
Main Results:
- Overexpression of YAP or TAZ significantly increased CTGF expression in ARPE-19 cells.
- Both K-975 and luteolin treatment attenuated the increased CTGF expression.
- TGF-β2 treatment upregulated YAP, TAZ, and CTGF, an effect reduced by K-975 and luteolin.
Conclusions:
- YAP and TAZ are involved in the regulation of CTGF expression in ARPE-19 cells.
- K-975 and luteolin demonstrate potential as therapeutic agents to inhibit CTGF production.
- These findings suggest a novel therapeutic avenue for preventing CTGF-driven vitreoretinal fibrosis.
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