The complement system forms the central core of innate immunity but also mediates a variety of inflammatory diseases. Recent studies with human subjects and animal models demonstrated that complement component C3a plays a novel and important role in the pathogenesis of asthma. The cellular and molecular mechanism by which C3a modulates asthma, however, remains unknown. Our preliminary studies demonstrated that G protein coupled receptor (GPCR) for C3a (C3aR) are expressed in human mast cells and that C3a causes rapid mast cell degranulation/leukotriene C4 generation and delayed NF-?B activation/chemokine generation;pathways that coordinately regulate airway hyperresponsiveness (AHR) and lung inflammation, two major features of asthma. Our preliminary studies also suggest new concepts in the regulation of C3aR function in mast cells, which may provide novel targets for differential regulation of AHR and lung inflammation. Receptor phosphorylation by G protein coupled receptor kinases (GRKs) and the subsequent recruitment of ?-arrestin provides an important mechanism for desensitization of GPCRs. We made the novel observation that C3aR phosphorylation, which inhibits mast cell degranulation, provides a stimulatory signal for chemokine gene expression. Based on these findings, we hypothesize that C3aR phosphorylation by distinct GRKs differentially regulates AHR and lung inflammation by inhibiting mast cell degranulation but stimulating chemokine gene expression.
Three specific aims are proposed to test this hypothesis.
In aim #1, we will test the hypothesis that phosphorylation of C3aR at distinct site by different GRKs inhibits mast cell degranulation but promotes NF-?B activation/chemokine generation.
In aim #2, we will test the hypothesis that phosphorylated C3aR associates with both 2-arrestin and PSD-95/Dlg/Zo1 (PDZ) domain containing proteins to inhibit and stimulate NF-?B activity, respectively.
In aim #3, we will modulate allergen-induced AHR and lung inflammation in vivo by targeting C3aR phosphorylation in mast cells. Collectively, these studies will generate significant new information on how C3aR signaling modulates asthma and may offer novel therapeutic approaches for the treatment of asthma and airway inflammation. Narrative: Asthma is a complex airway inflammatory disease characterized by bronchoconstriction, airway hyperresponsiveness (AHR) and inflammation. Approximately 17 million Americans are estimated to have asthma, one third of them children. In recent years, asthma prevalence and severity have been increasing dramatically world-wide. Mast cells present in the lung release mediators that cause the symptoms of asthma. This proposal is based on the identification of a new molecule that regulates mast cell function in asthma. We believe that proposed studies will generate significant new information on the regulation of mast cells and may offer novel therapeutic approaches for the treatment of asthma.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL085774-03
Application #
7848270
Study Section
Cellular and Molecular Immunology - A Study Section (CMIA)
Program Officer
Noel, Patricia
Project Start
2008-09-20
Project End
2012-06-01
Budget Start
2010-06-02
Budget End
2011-06-01
Support Year
3
Fiscal Year
2010
Total Cost
$393,750
Indirect Cost
Name
University of Pennsylvania
Department
Pathology
Type
Schools of Dentistry
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Subramanian, Hariharan; Gupta, Kshitij; Ali, Hydar (2016) Roles of Mas-related G protein-coupled receptor X2 on mast cell-mediated host defense, pseudoallergic drug reactions, and chronic inflammatory diseases. J Allergy Clin Immunol 138:700-710
Subramanian, Hariharan; Gupta, Kshitij; Parameswaran, Narayanan et al. (2014) Regulation of Fc?RI signaling in mast cells by G protein-coupled receptor kinase 2 and its RH domain. J Biol Chem 289:20917-27
Subramanian, Hariharan; Gupta, Kshitij; Lee, Donguk et al. (2013) ?-Defensins activate human mast cells via Mas-related gene X2. J Immunol 191:345-52
Gupta, Kshitij; Subramanian, Hariharan; Klos, Andreas et al. (2012) Phosphorylation of C3a receptor at multiple sites mediates desensitization, ?-arrestin-2 recruitment and inhibition of NF-?B activity in mast cells. PLoS One 7:e46369
Guo, Qiang; Subramanian, Hariharan; Gupta, Kshitij et al. (2011) Regulation of C3a receptor signaling in human mast cells by G protein coupled receptor kinases. PLoS One 6:e22559
Subramanian, Hariharan; Gupta, Kshitij; Guo, Qiang et al. (2011) Mas-related gene X2 (MrgX2) is a novel G protein-coupled receptor for the antimicrobial peptide LL-37 in human mast cells: resistance to receptor phosphorylation, desensitization, and internalization. J Biol Chem 286:44739-49
Vibhuti, Arpana; Gupta, Kshitij; Subramanian, Hariharan et al. (2011) Distinct and shared roles of ?-arrestin-1 and ?-arrestin-2 on the regulation of C3a receptor signaling in human mast cells. PLoS One 6:e19585
Kashem, Sakeen W; Subramanian, Hariharan; Collington, Sarah J et al. (2011) G protein coupled receptor specificity for C3a and compound 48/80-induced degranulation in human mast cells: roles of Mas-related genes MrgX1 and MrgX2. Eur J Pharmacol 668:299-304
Subramanian, Hariharan; Kashem, Sakeen W; Collington, Sarah J et al. (2011) PMX-53 as a dual CD88 antagonist and an agonist for Mas-related gene 2 (MrgX2) in human mast cells. Mol Pharmacol 79:1005-13
Ali, Hydar (2010) Regulation of human mast cell and basophil function by anaphylatoxins C3a and C5a. Immunol Lett 128:36-45