Toxoplasma gondii chronically infects approximately 30% of the USA population and causes severe life-threatening Toxoplasmic encephalitis infections in AIDS. Current drug treatments are not well tolerated, and because these treatments have little effect on the slow growing bradyzoite stages, latent parasites remain as a source of recrudescing infection in AIDS. More effective therapies that can target both tachyzoite and bradyzoite stages are urgently needed to treat acute infections as well as to prevent the toxoplasmosis caused by recrudescing parasites. Our preliminary studies have demonstrated that disruption of carbamoyl phosphate synthetase II (CPSII) in the de novo pyrimidine synthesis pathway causes a severe uracil auxotrophy with a corresponding loss of parasite growth and virulence. Consequently the pyrimidine biosynthetic pathway appears to be an excellent target for intervention, and fundamental studies on pyrimidine acquisition have the potential to identify new targets. Due to the need for uridine monophosphate in all life stages, we hypothesize that pyrimidine biosynthesis is necessary to sustain rapid tachyzoite replication as well as the viability of bradyzoites within tissue cysts. We propose to use a combination of genetic, biochemical, and cell biological approaches to better understand the fundamental biology of de novo pyrimidine synthesis and salvage pathways in T. gondii. Inhibitors of pyrimidine biosynthesis will be identified based on a screening strategy of pyrimidine auxotrophy. The information and reagents developed in our studies will be used to validate significant drug targets in the pathway as well as to validate the target(s) of inhibitors identified in this project. These studies will contribute to the development of new therapeutic agents to treat acute infections and may contribute to the development of the first therapeutic agents capable of clearing chronic infection to eliminate reactivation Toxoplasmic encephalitis in AIDS.

Public Health Relevance

This research project will validate drug targets and identify inhibitors of pyrimidine biosynthesis in Toxoplasma gondii using fundamental genetic, biochemical, and drug screening approaches. Consequently, this research project will accelerate the discovery of new vaccines and treatments for devastating opportunistic parasite infections in AIDS.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI041930-15
Application #
8441604
Study Section
AIDS-associated Opportunistic Infections and Cancer Study Section (AOIC)
Program Officer
Rogers, Martin J
Project Start
1997-08-01
Project End
2014-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
15
Fiscal Year
2013
Total Cost
$363,912
Indirect Cost
$133,588
Name
Dartmouth College
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
041027822
City
Hanover
State
NH
Country
United States
Zip Code
03755
Konradt, Christoph; Ueno, Norikiyo; Christian, David A et al. (2016) Endothelial cells are a replicative niche for entry of Toxoplasma gondii to the central nervous system. Nat Microbiol 1:
Fox, Barbara A; Rommereim, Leah M; Guevara, Rebekah B et al. (2016) The Toxoplasma gondii Rhoptry Kinome Is Essential for Chronic Infection. MBio 7:
Hortua Triana, Miryam Andrea; Cajiao Herrera, Daniela; Zimmermann, Barbara H et al. (2016) Pyrimidine Pathway-Dependent and -Independent Functions of the Toxoplasma gondii Mitochondrial Dihydroorotate Dehydrogenase. Infect Immun 84:2974-81
Sanders, Kiah L; Fox, Barbara A; Bzik, David J (2015) Attenuated Toxoplasma gondii Stimulates Immunity to Pancreatic Cancer by Manipulation of Myeloid Cell Populations. Cancer Immunol Res 3:891-901
Fox, Barbara A; Bzik, David J (2015) Nonreplicating, cyst-defective type II Toxoplasma gondii vaccine strains stimulate protective immunity against acute and chronic infection. Infect Immun 83:2148-55
Zhao, Yanlin; Marple, Andrew H; Ferguson, David J P et al. (2014) Avirulent strains of Toxoplasma gondii infect macrophages by active invasion from the phagosome. Proc Natl Acad Sci U S A 111:6437-42
Mouveaux, Thomas; Oria, Gabrielle; Werkmeister, Elisabeth et al. (2014) Nuclear glycolytic enzyme enolase of Toxoplasma gondii functions as a transcriptional regulator. PLoS One 9:e105820
Patil, Veerupaxagouda; Zhao, Yanlin; Shah, Suhagi et al. (2014) Co-existence of classical and alternative activation programs in macrophages responding to Toxoplasma gondii. Int J Parasitol 44:161-4
Baird, Jason R; Byrne, Katelyn T; Lizotte, Patrick H et al. (2013) Immune-mediated regression of established B16F10 melanoma by intratumoral injection of attenuated Toxoplasma gondii protects against rechallenge. J Immunol 190:469-78
Fox, Barbara A; Sanders, Kiah L; Bzik, David J (2013) Non-replicating Toxoplasma gondii reverses tumor-associated immunosuppression. Oncoimmunology 2:e26296

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