G proteins 23 subunits play a central role in G-protein coupled receptor (GPCR)- mediated signal transduction. They act as cofactors in the receptor-mediated activation process as well as playing direct roles in signal transfer to downstream targets. Considerable data has accumulated in number of systems that excess 23 signaling has pathological consequences and that manipulation of 23 subunit signaling could be an effective therapeutic strategy in heart failure as well as other diseases. We have developed a novel targeting strategy for selective manipulation of G protein 23 subunit signaling pathways by selectively blocking 23-subunit binding interactions with functional protein partners using small molecules. In our preliminary data we demonstrate these compounds are efficacious in animal models of heart failure, inflammation and morphine-dependent analgesia. In the proposed experiments we will explore the fundamental mechanisms underlying binding and selectivity of these 23 binding compounds.
In Specific aim 1 the experiments will characterize the selectivity of compounds for a broad range of G23 targets and develop new assays for predicting compound selectivity.
Specific aim 2 will combine biophysical ligand interaction methods, mutagenesis and x-ray crystallography to develop a detailed understanding of the mechanisms for selectivity.
Specific aim3 will exploit the information from specific aim 2 to refine our computational screening approach to allow for more efficient compound identification that may have unique mechanisms of action.
Specific aim 4 will apply our knowledge of selectivity to GPCR signaling in neutrophil functions. Successful completion of the proposed experiments will lead to a thorough understanding of a the mechanism of action of a new family of molecules that target G23 signaling that have potential uses in dissecting the mechanisms of action of GPCR stimulated signaling and providing the basis for novel therapeutic approaches. G protein coupled receptors (GPCRs) are a major class of transmembrane receptors responsible for recognition of a large class of diverse ligands. Here we propose investigation of selective small molecule inhibitors of G protein 23 subunits identified in our laboratory which could be used to inhibit multiple GPCRs and modify actions of existing GPCR directed pharmaceuticals. Public Health Relevance: Results of these experiments will help to validate this alternate approach to modification of signaling pathways downstream of GPCRs that could ultimately lead to development of novel therapeutics.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM081772-03
Application #
7755402
Study Section
Special Emphasis Panel (ZRG1-MIST-G (01))
Program Officer
Dunsmore, Sarah
Project Start
2008-04-01
Project End
2012-01-31
Budget Start
2010-02-01
Budget End
2011-01-31
Support Year
3
Fiscal Year
2010
Total Cost
$304,920
Indirect Cost
Name
University of Rochester
Department
Pharmacology
Type
Schools of Dentistry
DUNS #
041294109
City
Rochester
State
NY
Country
United States
Zip Code
14627
Rangel-Moreno, Javier; To, Jesi Y; Owen, Teresa et al. (2016) Inhibition of G Protein ?? Subunit Signaling Abrogates Nephritis in Lupus-Prone Mice. Arthritis Rheumatol 68:2244-56
Surve, Chinmay R; To, Jesi Y; Malik, Sundeep et al. (2016) Dynamic regulation of neutrophil polarity and migration by the heterotrimeric G protein subunits G?i-GTP and G??. Sci Signal 9:ra22
Brand, Cameron S; Sadana, Rachna; Malik, Sundeep et al. (2015) Adenylyl Cyclase 5 Regulation by G?? Involves Isoform-Specific Use of Multiple Interaction Sites. Mol Pharmacol 88:758-67
Smrcka, Alan V (2015) Regulation of phosphatidylinositol-specific phospholipase C at the nuclear envelope in cardiac myocytes. J Cardiovasc Pharmacol 65:203-10
Malik, S; deRubio, R G; Trembley, M et al. (2015) G protein ?? subunits regulate cardiomyocyte hypertrophy through a perinuclear Golgi phosphatidylinositol 4-phosphate hydrolysis pathway. Mol Biol Cell 26:1188-98
Kamal, Fadia A; Mickelsen, Deanne M; Wegman, Katherine M et al. (2014) Simultaneous adrenal and cardiac g-protein-coupled receptor-g?? inhibition halts heart failure progression. J Am Coll Cardiol 63:2549-2557
Le, Nhat-Tu; Takei, Yuichiro; Izawa-Ishizawa, Yuki et al. (2014) Identification of activators of ERK5 transcriptional activity by high-throughput screening and the role of endothelial ERK5 in vasoprotective effects induced by statins and antimalarial agents. J Immunol 193:3803-15
Surve, Chinmay R; Lehmann, David; Smrcka, Alan V (2014) A chemical biology approach demonstrates G protein ?? subunits are sufficient to mediate directional neutrophil chemotaxis. J Biol Chem 289:17791-801
Sun, Zhizeng; Smrcka, Alan V; Chen, Songhai (2013) WDR26 functions as a scaffolding protein to promote G??-mediated phospholipase C ?2 (PLC?2) activation in leukocytes. J Biol Chem 288:16715-25
Smrcka, Alan V (2013) Molecular targeting of G? and G?? subunits: a potential approach for cancer therapeutics. Trends Pharmacol Sci 34:290-8

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