This proposal seeks to continue a successful line of investigation into how skin and microbial proteases modify antimicrobial peptide function. Previous data have shown that activation of cathelicidin antimicrobial peptides is dependent on serine proteases including Kallikrein 5 and 7. Alternative processing of the substrate cathelicidin precursor hCAP 18 results in a variety of mature peptides with alternative function. Control of this processing is relevant to skin diseases ie. rosacea and psoriasis. Additional processing by microbial proteases serves as a virulence mechanism to alter normal host immunity. Recent data have shown alternative enzymatic processing of cathelicidin enables keratinocytes to recognize self-DNA through TLR9 and that TLR9 is essential for defense against skin infection by Group A Streptococcus. These findings suggest a novel role for nucleic acid recognition by skin epithelium. The focus of our proposal is to define what proteolytic products of cathelicidin processing control pattern. This approach will also extend to evaluation of how microbes influence these events.
Specific Aim 1. Define the structure-function relationships between cathelicidin peptides and the capacity of keratinocytes to respond to CpG and genomic DNA. We will generate a library of cathelicidin peptides and test their ability to influence TLR9 activation. This screen will define the critical structural elements of the peptide and identify key enzymatic steps that result in the generation of these products.
Specific Aim 2. Characterize the immune response to products defined in SA1 to determine the consequences of this to skin inflammation. We will establish the physiological relevance of peptides identified in aim 1 by using mouse models of skin inflammation, examine how the products of keratinocytes stimulated by alternative cathelicidin peptides alter T cell polarization, and establish the role of these findings in the in vivo skin inflammatory response.
Specific Aims3. Determine how protease and nuclease expression by a bacterial pathogen modulates host cutaneous innate immune responses. We will study how protease expression by a bacterial pathogen changes the size and activity profile of cathelicidin peptides in skin. We will investigate cathelicidin expression in DNA-based extracellular traps (ETs) produced by neutrophils and mast cells and their effect on keratinocyte TLR-9 signaling. We will study how bacterial protease and nuclease expression influences these interactions, and how cathelicidins themselves may promote ET production.

Public Health Relevance

The action of enzymes influences immune function by changing the way components of the immune system function. We have discovered that peptides produced by skin cells require specific enzymes for activation. This proposal will study in detail how specific peptide and nucleic acid products of enzymes control skin inflammation and influence human skin diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR052728-09
Application #
8607896
Study Section
Arthritis, Connective Tissue and Skin Study Section (ACTS)
Program Officer
Cibotti, Ricardo
Project Start
2005-07-01
Project End
2016-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
9
Fiscal Year
2014
Total Cost
$313,875
Indirect Cost
$111,375
Name
University of California San Diego
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
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Cole, Jason N; Nizet, Victor (2016) Bacterial Evasion of Host Antimicrobial Peptide Defenses. Microbiol Spectr 4:
Kumaraswamy, Monika; Lin, Leo; Olson, Joshua et al. (2016) Standard susceptibility testing overlooks potent azithromycin activity and cationic peptide synergy against MDR Stenotrophomonas maltophilia. J Antimicrob Chemother 71:1264-9
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Borkowski, Andrew W; Kuo, I-Hsin; Bernard, Jamie J et al. (2015) Toll-like receptor 3 activation is required for normal skin barrier repair following UV damage. J Invest Dermatol 135:569-78
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MacLeod, Daniel T; Nakatsuji, Teruaki; Wang, Zhenping et al. (2015) Vaccinia virus binds to the scavenger receptor MARCO on the surface of keratinocytes. J Invest Dermatol 135:142-50
Beaumont, Paula E; McHugh, Brian; Gwyer Findlay, Emily et al. (2014) Cathelicidin host defence peptide augments clearance of pulmonary Pseudomonas aeruginosa infection by its influence on neutrophil function in vivo. PLoS One 9:e99029
Santos, José Carlos; Silva-Gomes, Sandro; Silva, João Pedro et al. (2014) Endogenous cathelicidin production limits inflammation and protective immunity to Mycobacterium avium in mice. Immun Inflamm Dis 2:1-12

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