Effect of Brd4 Inhibition on the Expression of Pro-Fibrotic Markers
College
The Graduate School
Department
Department of Biomedical and Translational Sciences
Graduate Level
Master’s
Graduate Program/Concentration
Eastern Virginia Medical School - Biomedical Sciences Research
Presentation Type
Poster Presentation
Abstract
Effect of Brd4 Inhibition on the Expression of Pro-Fibrotic Markers
Ryan Washington, Jennifer Zhou
Eastern Virginia Medical School
INTRODUCTION: Idiopathic Pulmonary Fibrosis (IPF) is a chronic lung disease characterized by excessive scarring of lung tissues. 80,380 Americans are affected by IPF today. The likelihood of developing the disease increases as getting older. IPF patients have increased activated fibroblasts compared to healthy people. Inhibition of Brd4, a transcription factor that regulates the transformation of fibroblasts into myofibroblasts has been proposed as a way to slow the progression of IPF. In this study, we utilized two Brd4 inhibitors, dBET6 and A1874, to test the effects of Brd4 inhibition on fibroblast activation.
METHODS: IMR90 cells were cultured in a 6-well plate in 2 mL of DMEM media in each well. The cells were incubated in serum-free media overnight to arrest them in G0. 24 hours later, the dishes were subdivided into 2 control wells, 2 wells that contained TGF-B, one well containing TGF-B and dBET6, and one well containing TGF-B and A1874. The plates were incubated for 24 hrs, then protein was extracted. Afterwards, Western blot Analysis was conducted to detect differences in protein expression.
RESULTS: We observed an increase in the expression of pro-fibrotic markers in the cells treated with TGF-B with or without inhibitors when compared to control. The cells treated with TGF-β and dBET6 showed lower levels of profibrotic markers (α-SMA and Col3A1) expression compared to the control. However, the cells that were treated with TGF-β /A1874 showed no detectable difference.
CONCLUSION: In this study, we demonstrated that dBET6 decreases the expression of proteins associated with fibrosis in IMR90 cells induced by profibrotic cytokine TGF-β. These results suggest that dBET6 may play a protective role in the pathogenesis of IPF. In the future, further studies are needed to confirm the results in IPF patient samples.
Effect of Brd4 Inhibition on the Expression of Pro-Fibrotic Markers
Effect of Brd4 Inhibition on the Expression of Pro-Fibrotic Markers
Ryan Washington, Jennifer Zhou
Eastern Virginia Medical School
INTRODUCTION: Idiopathic Pulmonary Fibrosis (IPF) is a chronic lung disease characterized by excessive scarring of lung tissues. 80,380 Americans are affected by IPF today. The likelihood of developing the disease increases as getting older. IPF patients have increased activated fibroblasts compared to healthy people. Inhibition of Brd4, a transcription factor that regulates the transformation of fibroblasts into myofibroblasts has been proposed as a way to slow the progression of IPF. In this study, we utilized two Brd4 inhibitors, dBET6 and A1874, to test the effects of Brd4 inhibition on fibroblast activation.
METHODS: IMR90 cells were cultured in a 6-well plate in 2 mL of DMEM media in each well. The cells were incubated in serum-free media overnight to arrest them in G0. 24 hours later, the dishes were subdivided into 2 control wells, 2 wells that contained TGF-B, one well containing TGF-B and dBET6, and one well containing TGF-B and A1874. The plates were incubated for 24 hrs, then protein was extracted. Afterwards, Western blot Analysis was conducted to detect differences in protein expression.
RESULTS: We observed an increase in the expression of pro-fibrotic markers in the cells treated with TGF-B with or without inhibitors when compared to control. The cells treated with TGF-β and dBET6 showed lower levels of profibrotic markers (α-SMA and Col3A1) expression compared to the control. However, the cells that were treated with TGF-β /A1874 showed no detectable difference.
CONCLUSION: In this study, we demonstrated that dBET6 decreases the expression of proteins associated with fibrosis in IMR90 cells induced by profibrotic cytokine TGF-β. These results suggest that dBET6 may play a protective role in the pathogenesis of IPF. In the future, further studies are needed to confirm the results in IPF patient samples.