Pseudomonas aeruginosa (P. aeruginosa) is a Gram-negative, opportunistic pathogen associated with bacterial keratitis, especially in contact lens usage. With the increasing incidence of antibiotic resistance, our goal is to determine the role of the pro- inflammatory neuropeptide, Substance P (SP) and the anti-inflammatory neuropeptide, vasoactive intestinal peptide (VIP) in regulation of inflammation, innate immunity and restoration of corneal homeostasis.
The aims of this proposal are: 1) to test the hypothesis that VIP modulates TLRs and TLR-related adaptor molecules;down- regulates the expression of adhesion molecules, decreases the migration/persistence of inflammatory cells (PMN, M?, Langerhans cells) in the infected cornea;and promotes restoration of corneal homeostasis and re-modeling of the ECM, 2) to test the hypothesis that SP exacerbates disease pathogenesis through the up-regulation of TLRs and TLR- related adaptor molecules;up-regulates adhesion molecule expression and enhances the migration/persistence of inflammatory cells (PMN, M?, Langerhans cells) in the infected cornea and degradation of the ECM. Experiments will include use of techniques such as real-time RT-PCR, short interfering RNA (siRNA), as well as plate counts, enzyme linked immunosorbant assay (ELISA), myeloperoxidase assay (MPO) for neutrophil (PMN) quantitation, dual antibody immunostaining, histopathology, and Western blotting. Our findings will be particularly significant for development of novel therapeutics for bacterial keratitis, targeting not only regulation of corneal infection and inflammation, but also the critical aspect of promoting tissue repair. Since in the United States alone the incidence of microbial keratitis is 25,000-30,000 cases annually with cost of treatment estimated at $15-30 million, the studies are relevant to human health and have considerable medical and economic impact.

Public Health Relevance

P. aeruginosa is a Gram-negative pathogen associated with bacterial keratitis, particularly in contact lens wearers, with considerable medical and economic consequences. Understanding the mechanisms of neuropeptide regulation of Toll-like receptor and associated adaptor molecule activation and modulation of adhesion molecules and extracellular matrix components will provide substantive insight and novel therapeutic potential to regulate disease pathogenesis in cornea and perhaps other infectious and immune-based inflammatory diseases.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY002986-34
Application #
8374119
Study Section
Anterior Eye Disease Study Section (AED)
Program Officer
Mckie, George Ann
Project Start
1979-04-01
Project End
2014-11-30
Budget Start
2012-12-01
Budget End
2014-11-30
Support Year
34
Fiscal Year
2013
Total Cost
$408,451
Indirect Cost
$139,733
Name
Wayne State University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
001962224
City
Detroit
State
MI
Country
United States
Zip Code
48202
Li, Cui; McClellan, Sharon A; Barrett, Ronald et al. (2014) Interleukin 17 regulates Mer tyrosine kinase-positive cells in Pseudomonas aeruginosa keratitis. Invest Ophthalmol Vis Sci 55:6886-900
Hazlett, Linda D; Jiang, Xiaoyu; McClellan, Sharon A (2014) IL-10 function, regulation, and in bacterial keratitis. J Ocul Pharmacol Ther 30:373-80
Jiang, Xiaoyu; McClellan, Sharon A; Barrett, Ronald et al. (2014) HGF signaling impacts severity of Pseudomonas aeruginosa keratitis. Invest Ophthalmol Vis Sci 55:2180-90
Berger, Elizabeth A; McClellan, Sharon A; Vistisen, Kerry S et al. (2013) HIF-1* is essential for effective PMN bacterial killing, antimicrobial peptide production and apoptosis in Pseudomonas aeruginosa keratitis. PLoS Pathog 9:e1003457
Foldenauer, Megan E B; McClellan, Sharon A; Berger, Elizabeth A et al. (2013) Mammalian target of rapamycin regulates IL-10 and resistance to Pseudomonas aeruginosa corneal infection. J Immunol 190:5649-58
Jiang, Xiaoyu; McClellan, Sharon A; Barrett, Ronald P et al. (2011) VIP and growth factors in the infected cornea. Invest Ophthalmol Vis Sci 52:6154-61
Berger, Elizabeth A; McClellan, Sharon A; Barrett, Ronald P et al. (2011) Testican-1 promotes resistance against Pseudomonas aeruginosa-induced keratitis through regulation of MMP-2 expression and activation. Invest Ophthalmol Vis Sci 52:5339-46
Hazlett, Linda D; Hendricks, Robert L (2010) Reviews for immune privilege in the year 2010: immune privilege and infection. Ocul Immunol Inflamm 18:237-43
Berger, Elizabeth A; McClellan, Sharon A; Barrett, Ronald P et al. (2010) VIP promotes resistance in the Pseudomonas aeruginosa-infected cornea by modulating adhesion molecule expression. Invest Ophthalmol Vis Sci 51:5776-82
Zhou, Zimei; Wu, Minhao; Barrett, Ronald P et al. (2010) Role of the Fas pathway in Pseudomonas aeruginosa keratitis. Invest Ophthalmol Vis Sci 51:2537-47

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