This is a revised grant application to study global gene expression in Cryptosporidium parvum and in host cells infected with this parasite. Cryptosporidium parvum is an opportunistic pathogen in people with AIDS and is a common cause of diarrhea in children worldwide. The completed sequence of the C. parvum genome provides new opportunities for identifying drug targets and for research on this parasite on a genomic and functional level. In this project DNA microarrays will be used to study parasite gene regulation and the response of the host cell to this infection. Analysis of global parasite transcription, primarily during excystation, host cell invasion, and the initial phase of intracellular development will identify parasite biochemical pathways, which are differentially expressed during this phase of the life cycle. Analysis of the transcriptional response of infected host cells in culture and in animal models will identify mechanisms by which the host responds to the infection, and shed light on the molecular pathways leading to the intestinal symptoms of cryptosporidosis. This project is a collaborative effort between two research groups with complementary expertise in the biology and genomics of C. parvum and extensive collaborative experience.
In Specific Aim 1 a comprehensive DNA microarray containing all of the genes of C. parvum will be developed. This task will take advantage of the genomic sequence by converting genomic information into a tool for identifying genetic processes associated with parasite development.
In Specific Aim 2 microarrays developed under aim 1 will be used to probe global gene expression during oocyst excystation, host cell invasion, and first generation merogony.
Specific Aim 3 will use commercially available human and mouse gene arrays to study the response of the host cell to C. parvum. A comparison of the host cell response to C. parvum type 1 and type 2 will be performed to probe the molecular basis of the observed difference in virulence between types.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI055347-01A2
Application #
6841097
Study Section
Special Emphasis Panel (ZRG1-AARR-B (03))
Program Officer
Rogers, Martin J
Project Start
2004-07-01
Project End
2008-06-30
Budget Start
2004-07-01
Budget End
2005-06-30
Support Year
1
Fiscal Year
2004
Total Cost
$446,655
Indirect Cost
Name
Tufts University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
039318308
City
Boston
State
MA
Country
United States
Zip Code
02111
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Mazurie, Aurelien; Bonchev, Danail; Schwikowski, Benno et al. (2010) Evolution of metabolic network organization. BMC Syst Biol 4:59
Vanee, Niti; Roberts, Seth B; Fong, Stephen S et al. (2010) A genome-scale metabolic model of Cryptosporidium hominis. Chem Biodivers 7:1026-39
Cohn, Benjamin; Manque, Patricio; Lara, Ana M et al. (2010) Putative cis-regulatory elements associated with heat shock genes activated during excystation of Cryptosporidium parvum. PLoS One 5:e9512
Yang, Yi-Lin; Buck, Gregory A; Widmer, Giovanni (2010) Cell sorting-assisted microarray profiling of host cell response to Cryptosporidium parvum infection. Infect Immun 78:1040-8
Widmer, G (2009) Meta-analysis of a polymorphic surface glycoprotein of the parasitic protozoa Cryptosporidium parvum and Cryptosporidium hominis. Epidemiol Infect 137:1800-8
Zhang, L; Sheoran, A S; Widmer, G (2009) Cryptosporidium parvum DNA replication in cell-free culture. J Parasitol 95:1239-42
Valente, Andre X C N; Roberts, Seth B; Buck, Gregory A et al. (2009) Functional organization of the yeast proteome by a yeast interactome map. Proc Natl Acad Sci U S A 106:1490-5
Roberts, Seth B; Robichaux, Jennifer L; Chavali, Arvind K et al. (2009) Proteomic and network analysis characterize stage-specific metabolism in Trypanosoma cruzi. BMC Syst Biol 3:52
Yang, Yi-Lin; Serrano, Myrna G; Sheoran, Abhineet S et al. (2009) Over-expression and localization of a host protein on the membrane of Cryptosporidium parvum infected epithelial cells. Mol Biochem Parasitol 168:95-101

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