. The pathogenic fungus Cryptococcus neoformans causes life-threatening infections in AIDS patients and therefore poses a major threat to the >34 million people worldwide who are infected with HIV. The related species Cryptococcus gattii has recently emerged as a primary pathogen of immunocompetent people. The long-term goal of this project is to acquire knowledge that will lead to new strategies to combat fungal infections. In particular, we want to establish a detailed understanding of the factors required for pathogen growth in mammalian hosts and identify useful targets for therapy. In this regard, iron availability is a key indicator of the host environment as well as an essential nutrient for pathogen proliferation. In addition, mammals actively withhold iron from pathogens in a process termed nutritional immunity, and pathogens must therefore aggressively compete for iron. Iron is particularly important for the pathogenesis of C. neoformans because the availability of this metal not only influences growth but also the size of the polysaccharide capsule that is the major virulence factor. To fill gaps in our understanding of the mechanisms by which fungal pathogens compete for iron, the first specific aim is to determine the molecular functions of cell surface proteins that support iron acquisition from heme. These proteins each contribute to robust growth on heme and include a secreted mannoprotein that is a candidate heme-binding protein as well as three ferric reductases. A second specific aim will characterize the intracellular machinery for heme trafficking and processing. An insertional mutagenesis screen identified 25 genes in which mutations caused growth defects on heme, and some of these genes encode intracellular trafficking machinery. This result led to the hypothesis that heme is acquired by endocytosis and transported to the vacuole for iron extraction and recycling. Mutants with defects at specific steps in known and candidate transport functions will be constructed and tested for their ability to process heme. A final specific aim will evaluate the role of heme and iron acquisition functions in the virulence of C. neoformans. Mutants lacking combinations of iron acquisition functions will be tested in mouse inhalation models of cryptococcosis and the proliferation of the mutants in macrophages will be examined in the context of different iron sources. These studies will provide a comprehensive view of the relative importance of specific host iron sources and fungal uptake strategies during cryptococcosis. !

Public Health Relevance

of this project comes from the pressing need to control fungal infections in humans with impaired immune systems. In particular, the 34 million or more people infected with HIV have a high chance of succumbing to fungal diseases. The research will specifically examine the potential to control these infections by targeting the abiliy of fungal pathogens to acquire iron as a vital nutrient during infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI053721-11
Application #
8616150
Study Section
Pathogenic Eukaryotes Study Section (PTHE)
Program Officer
Duncan, Rory A
Project Start
2003-08-01
Project End
2019-02-28
Budget Start
2014-03-15
Budget End
2015-02-28
Support Year
11
Fiscal Year
2014
Total Cost
$283,793
Indirect Cost
$21,022
Name
University of British Columbia
Department
Type
DUNS #
251949962
City
Vancouver
State
BC
Country
Canada
Zip Code
V6 1-Z3
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Do, Eunsoo; Hu, Guanggan; Caza, Mélissa et al. (2016) The ZIP family zinc transporters support the virulence of Cryptococcus neoformans. Med Mycol 54:605-15
Jung, Won Hee; Sham, Anita; White, Rick et al. (2016) Correction: Iron Regulation of the Major Virulence Factors in the AIDS-Associated Pathogen Cryptococcus neoformans. PLoS Biol 14:e1002410
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Caza, Mélissa; Hu, Guanggan; Price, Michael et al. (2016) The Zinc Finger Protein Mig1 Regulates Mitochondrial Function and Azole Drug Susceptibility in the Pathogenic Fungus Cryptococcus neoformans. mSphere 1:
Ding, Hao; Mayer, François L; Sánchez-León, Eddy et al. (2016) Networks of fibers and factors: regulation of capsule formation in Cryptococcus neoformans. F1000Res 5:
Choi, Jaehyuk; Jung, Won Hee; Kronstad, James W (2015) The cAMP/protein kinase A signaling pathway in pathogenic basidiomycete fungi: Connections with iron homeostasis. J Microbiol 53:579-87
Do, Eunsoo; Hu, Guanggan; Caza, Mélissa et al. (2015) Leu1 plays a role in iron metabolism and is required for virulence in Cryptococcus neoformans. Fungal Genet Biol 75:11-9
Hu, Guanggan; Caza, Mélissa; Cadieux, Brigitte et al. (2015) The endosomal sorting complex required for transport machinery influences haem uptake and capsule elaboration in Cryptococcus neoformans. Mol Microbiol 96:973-92
Saikia, Sanjay; Oliveira, Debora; Hu, Guanggan et al. (2014) Role of ferric reductases in iron acquisition and virulence in the fungal pathogen Cryptococcus neoformans. Infect Immun 82:839-50

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