The long term objective of this proposal is a comprehensive analysis of virulence factors in pathogenic yeast required for colonization and persistence in the urinary tract. Candida glabrata and Candida albicans are the two major organisms responsible for funguria, as well as vaginal, oropharyngeal and systemic candidiasis. Little is known about the factors that allow Candida to colonize or persist in the urinary tract. Here, we will take a genetic approach to understanding Candida infections of the urinary tract. Unlike C. albicans, C. glabrata is a genetic approach to understanding Candida infections of the urinary tract. Unlike C. albicans, C. glabrata is haploid making it an excellent model for genetic analysis of virulence using already established genetic tools. We propose first to identify genes specifically involved in colonization and persistence in the urinary tract. We will use two different complementary global approaches. First, a variation of signature-tagged mutagenesis that we have established in C. glabrata will be used to screen a bank of . glabrata mutants for those that are unable to colonize and replicate in the bladder. Second, a library of genomic fragments fused to GFP will be constructed and screened for promotes specifically induced in the bladder or kidney but not under laboratory conditions These two global approaches will yield overlapping sets of genes relevant to the host interaction... Lastly, we will analyze in detail the role of any identified genes in urinary tract infections. Identified genes will first be prioritized for further study based on sequence homology. Mutants will be characterized with a set of in vitro assays for different aspects of the host interaction, including adherence to host tissue, interaction with phagocytes and growth under in vitro conditions mimicking the bladder environment. Different genes are likely to have roles in different aspects of infection which can be assayed only if the dynamics of the infection are known. We will therefore also use our signature-tagged strains to analyze the dynamics of UTI infection for wild type and mutant strains of C. glabrata, distinguishing between initial colonization and outgrowth. These approaches will provide insight into diverse aspects of the yeast- host interaction and an improved understanding of the processes contributing to fungal UTI, ultimately permitting enhanced therapeutic intervention.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Program Projects (P01)
Project #
5P01DK049720-07
Application #
6501062
Study Section
Special Emphasis Panel (ZDK1)
Project Start
2001-07-01
Project End
2002-06-30
Budget Start
Budget End
Support Year
7
Fiscal Year
2001
Total Cost
$129,815
Indirect Cost
Name
University of Maryland Baltimore
Department
Type
DUNS #
003255213
City
Baltimore
State
MD
Country
United States
Zip Code
21201
Buckles, Eric L; Luterbach, Courtney L; Wang, Xiaolin et al. (2015) Signature-tagged mutagenesis and co-infection studies demonstrate the importance of P fimbriae in a murine model of urinary tract infection. Pathog Dis 73:
Buckles, Eric L; Wang, Xiaolin; Lane, M Chelsea et al. (2009) Role of the K2 capsule in Escherichia coli urinary tract infection and serum resistance. J Infect Dis 199:1689-97
Lane, M Chelsea; Li, Xin; Pearson, Melanie M et al. (2009) Oxygen-limiting conditions enrich for fimbriate cells of uropathogenic Proteus mirabilis and Escherichia coli. J Bacteriol 191:1382-92
Zupancic, Margaret L; Frieman, Matthew; Smith, David et al. (2008) Glycan microarray analysis of Candida glabrata adhesin ligand specificity. Mol Microbiol 68:547-59
Jacobsen, Sandra M; Lane, Mary C; Harro, Jean M et al. (2008) The high-affinity phosphate transporter Pst is a virulence factor for Proteus mirabilis during complicated urinary tract infection. FEMS Immunol Med Microbiol 52:180-93
Ma, Biao; Pan, Shih-Jung; Zupancic, Margaret L et al. (2007) Assimilation of NAD(+) precursors in Candida glabrata. Mol Microbiol 66:14-25
Buckles, Eric L; Wang, Xiaolin; Lockatell, C Virginia et al. (2006) PhoU enhances the ability of extraintestinal pathogenic Escherichia coli strain CFT073 to colonize the murine urinary tract. Microbiology 152:153-60
Castano, Irene; Pan, Shih-Jung; Zupancic, Margaret et al. (2005) Telomere length control and transcriptional regulation of subtelomeric adhesins in Candida glabrata. Mol Microbiol 55:1246-58
Domergue, Renee; Castano, Irene; De Las Penas, Alejandro et al. (2005) Nicotinic acid limitation regulates silencing of Candida adhesins during UTI. Science 308:866-70
Jansen, Angela M; Lockatell, Virginia; Johnson, David E et al. (2004) Mannose-resistant Proteus-like fimbriae are produced by most Proteus mirabilis strains infecting the urinary tract, dictate the in vivo localization of bacteria, and contribute to biofilm formation. Infect Immun 72:7294-305

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