Pseudomonas aeruginosa infections result in significant morbidity and mortality in people suffering from cystic fibrosis (CF). A key feature of these infections is that they involve biofilm communities. Another is that during the course of chronic infection, P. aeruginosa acquires mutations that help it adapt to the CF environment. This proposal will investigate a clear link between biofilm formation and genetic adaptation, the RSCV phenotype. RSCVs generally exhibit increased biofilm formation and antimicrobial tolerance relative to the non- mucoid, smooth colony morphotype. One important trait of RSCVs are elevated production of the biofilm matrix exopolysaccharide PEL. The structure of PEL remains unknown, and its elucidation is a goal of this proposal. In addition, P. aeruginosa RSCVs are actively selected for during the course of chronic cystic fibrosis (CF) airway infections. We recently discovered that the prevalence of RSCVs in the CF airways has been grossly underestimated and we subsequently identified novel genetic loci linked to the RSCV phenotype. The molecular mechanism underpinning why these mutations produce RSCVs is unclear. Finally, we found that RSCVs can be highly variable phenotypically, depending upon the causal mutation. Some of these variable phenotypes are pathogenically relevant, suggesting that different RSCVs may vary in their ability to form biofilms and cause chronic infection. The overall goals of this application are to determine the structure of PEL and to elucidate the molecular mechanism leading to the RSCV phenotype that results from novel RSCV- linked mutations. We will also examine the pathogenic fitness advantages of different RSCV genotypes using chronic animal models of infection and in vitro cellular immunological assays. Once mucoid genetic variants of P. aeruginosa emerge, prognosis of CF patients clearly worsens. Thus, there exists a critical window of opportunity for immunological or chemotherapeutic intervention prior to mucoid conversion. RSCVs are usually observed prior to mucoidy, thus therapeutic strategies taking advantage of this window of opportunity would have to account for them. A thorough analysis of RSCVs and Pel may lead to the development of therapeutic agents and improve the quality of life for CF patients as well as individuals with other P. aeruginosa infections that involve biofilms.

Public Health Relevance

This proposal is focused on examining the role of RSCVs in Pseudomonas aeruginosa biofilm formation and pathogenesis as it relates to cystic fibrosis infection. There exists a critical window of opportunity for immunological or chemotherapeutic intervention in CF prior to mucoid conversion. RSCVs are usually observed prior to mucoidy, thus therapeutic strategies taking advantage of this window of opportunity would have to account for them.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI077628-05A1
Application #
8757461
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Taylor, Christopher E,
Project Start
2008-04-01
Project End
2019-06-30
Budget Start
2014-07-10
Budget End
2015-06-30
Support Year
5
Fiscal Year
2014
Total Cost
$450,187
Indirect Cost
$108,951
Name
University of Washington
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
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Zhao, Kun; Tseng, Boo Shan; Beckerman, Bernard et al. (2013) Psl trails guide exploration and microcolony formation in Pseudomonas aeruginosa biofilms. Nature 497:388-91
Marmont, Lindsey S; Whitney, John C; Robinson, Howard et al. (2012) Expression, purification, crystallization and preliminary X-ray analysis of Pseudomonas aeruginosa PelD. Acta Crystallogr Sect F Struct Biol Cryst Commun 68:181-4
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Colvin, Kelly M; Gordon, Vernita D; Murakami, Keiji et al. (2011) The pel polysaccharide can serve a structural and protective role in the biofilm matrix of Pseudomonas aeruginosa. PLoS Pathog 7:e1001264
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