Negative Regulators of Insulin Signaling Revealed in a Genome-Wide Functional Screen

被引:25
作者
Huang, Shih-Min A.
Hancock, Michael K.
Pitman, Jeffrey L.
Orth, Anthony P.
Gekakis, Nicholas
机构
[1] Genomics Institute of the Novartis Research Foundation, San Diego, CA
[2] The Scripps Research Institute, La Jolla, CA
[3] Novartis Institute for Biomedical Research, Developmental and Molecular Pathways, Cambridge, MA
[4] Invitrogen, Madison, WI
来源
PLOS ONE | 2009年 / 4卷 / 09期
关键词
D O I
10.1371/journal.pone.0006871
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: Type 2 diabetes develops due to a combination of insulin resistance and beta-cell failure and current therapeutics aim at both of these underlying causes. Several negative regulators of insulin signaling are known and are the subject of drug discovery efforts. We sought to identify novel contributors to insulin resistance and hence potentially novel targets for therapeutic intervention. Methodology: An arrayed cDNA library encoding 18,441 human transcripts was screened for inhibitors of insulin signaling and revealed known inhibitors and numerous potential novel regulators. The novel hits included proteins of various functional classes such as kinases, phosphatases, transcription factors, and GTPase associated proteins. A series of secondary assays confirmed the relevance of the primary screen hits to insulin signaling and provided further insight into their modes of action. Conclusion/Significance: Among the novel hits was PALD (KIAA1274, paladin), a previously uncharacterized protein that when overexpressed led to inhibition of insulin's ability to down regulate a FOXO1A-driven reporter gene, reduced upstream insulin-stimulated AKT phosphorylation, and decreased insulin receptor (IR) abundance. Conversely, knockdown of PALD gene expression resulted in increased IR abundance, enhanced insulin-stimulated AKT phosphorylation, and an improvement in insulin's ability to suppress FOXO1A-driven reporter gene activity. The present data demonstrate that the application of arrayed genome-wide screening technologies to insulin signaling is fruitful and is likely to reveal novel drug targets for insulin resistance and the metabolic syndrome.
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页数:9
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