A model to identify novel targets involved in oxidative stress-induced apoptosis in human lung epithelial cells by RNA interference.
Chronic obstructive pulmonary disease (COPD) is an increasing health problem primarily associated with cigarette smoking, and one of the leading causes of morbidity and mortality worldwide. Despite recent advances in understanding the pathogenesis of the disease, overall patient outcome remains poor with limited therapeutic intervention. Chronic inflammation, an imbalance between proteolytic and anti-proteolytic activities (leading to lung parenchyma destruction) and excessive oxidative stress contribute to COPD pathophysiology. Oxidative stress-triggered apoptosis of alveolar structural cells, including epithelial cells, may be an important event in the development of COPD. In this study, we developed a cell-based oxidative stress-induced apoptosis assay and performed a high-throughput screen (HTS) using a human druggable genome siRNA library. Our results have identified potential novel pathways (e.g. unfolded protein response, proteosomal activity) and targets (e.g. MAP3K14, HMGB2) that regulate the response of lung epithelial cells to oxidative stress. This assay has proven to be a useful tool for the identification of potential new therapeutic targets for lung disease.
Wixted
William E
WE
Respiratory Center for Excellence in Drug Discovery, GlaxoSmithKline R&D, King of Prussia, PA 19406, USA.
Kitson
Chris
C
Colebrook
Jayne C
JC
Roberts
Emma J
EJ
Fox
Steven M
SM
Kou
Jen P
JP
Li
Jun U
JU
López-Boado
Yolanda S
YS
D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004847:Epithelial Cells / Q000187:drug effects*; D005434:Flow Cytometry; D015723:Gene Library; D006801:Humans; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors / Q000633:toxicity; D008168:Lung / Q000166:cytology* / Q000187:drug effects; D016877:Oxidants / Q000633:toxicity; D018384:Oxidative Stress / Q000187:drug effects*; D034622:RNA Interference / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects; D014162:Transfection
D000975:Antioxidants; D016207:Cytokines; D016877:Oxidants; D006861:Hydrogen Peroxide; D020169:Caspases