Morbidity and mortality associated with Chagas disease in Latin America exceed better-known conditions such as malaria, tuberculosis, or AIDS. Millions of people are affected by this trypanosomiasis. No vaccines are available to prevent this disease and drug treatments have serious side effects and are not completely effective. Survival of Trypanosoma cruzi in the mammalian hosts depends on the parasite's ability to infect host cells, reproduce, and live in the blood of the host long enough to warrant its transmission through a bloodsucking insect vector.
Our aim i s to investigate these survival mechanisms and their role in the pathogenesis of T. cruzi infection, and shed light into potential ways to control the disease. Survival of T. cruzi trypomastigotes in the blood of the mammalian host and in the intestine of the vector depends in great part on their ability to tolerate dramatic changes in osmolarity during their circulation through the kidney medulla of the mammalian host (1,300-1,400 mOsm/Kg) or their passage through the rectum of the insect vector (1,000 mOsm/Kg), and we have found that polyphosphate (polyP) and the contractile vacuole complex (CVC) have en essential role in their survival mechanisms. Recent results from our laboratory suggest that polyP and/or pyrophosphate (PPi) could also be involved in the pathogenesis of T. cruzi infection. To reduce polyP levels we overexpressed a degradative enzyme that hydrolyzes both PPi and polyP resulting in a dramatic decrease of PPi/polyP. Mutant trypomastigotes overexpressing the enzyme did not produce detectable parasitemias and in several experiments all infected animals survived an otherwise lethal infection. Mice infected with parasites deficient in PPi/polyP failed to develop parasitemia after immunosuppression with cyclophosphamide strongly suggesting that infection had been completely cleared. Our results underscore an important role for PPi/polyP in the pathogenesis of T. cruzi infection. PolyP has been shown to act as a virulence factor in bacteria but little is known on its role in eukaryotic pathogens, besides its pro-coagulant and pro-inflammatory activities. Survival within the mammalian host also depends on the ability of T. cruzi to invade different host cells, escape from the parasitophorus vacuole and replicate intracellularly. T. cruzi trans-sialidases have been demonstrated to have essential roles in these mechanisms. However, the main obstacle in assessing the function of trans-sialidases is that knockout parasites were never obtained due to the large number of gene copies scattered through the genome. We have found that trans-sialidases traffic through the contractile vacuole in their way to the plasma membrane, and that disruption of this traffic by interfering with their passage through the CVC results in parasites devoid of these proteins in their surface. The study of this trafficking mechanism and of ways to interfere with this traffic will contribute to the understanding of the pathogenesis of T. cruzi infection.

Public Health Relevance

Morbidity and mortality associated with Chagas disease in Latin America exceed better-known conditions such as malaria, tuberculosis, or AIDS, and millions of people are affected by this trypanosomiasis with no vaccines available to prevent it and serious side effects of drug treatments. The study of metabolic pathways in these parasites that may be essential for their survival but may not find an equivalent counterpart in their host could make possible the development of specific inhibitors as possible means of controlling the parasites without damaging the hosts. We are investigating the role of polyphosphate and the traffic of trans-sialidases in the pathogenesis of Trypanosoma cruzi infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI107663-01A1
Application #
8650941
Study Section
Pathogenic Eukaryotes Study Section (PTHE)
Program Officer
Wali, Tonu M
Project Start
2014-09-03
Project End
2018-08-31
Budget Start
2014-09-03
Budget End
2015-08-31
Support Year
1
Fiscal Year
2014
Total Cost
$372,916
Indirect Cost
$122,916
Name
University of Georgia
Department
Public Health & Prev Medicine
Type
Organized Research Units
DUNS #
004315578
City
Athens
State
GA
Country
United States
Zip Code
30602
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Lander, Noelia; Chiurillo, Miguel A; Docampo, Roberto (2016) Genome Editing by CRISPR/Cas9: A Game Change in the Genetic Manipulation of Protists. J Eukaryot Microbiol 63:679-90
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Chao, María N; Matiuzzi, Carolina Exeni; Storey, Melissa et al. (2015) Aryloxyethyl Thiocyanates Are Potent Growth Inhibitors of Trypanosoma cruzi and Toxoplasma gondii. ChemMedChem 10:1094-108
Lander, Noelia; Li, Zhu-Hong; Niyogi, Sayantanee et al. (2015) CRISPR/Cas9-Induced Disruption of Paraflagellar Rod Protein 1 and 2 Genes in Trypanosoma cruzi Reveals Their Role in Flagellar Attachment. MBio 6:e01012
Veiga-Santos, Phercyles; Li, Kai; Lameira, Lilianne et al. (2015) SQ109, a new drug lead for Chagas disease. Antimicrob Agents Chemother 59:1950-61
Niyogi, Sayantanee; Docampo, Roberto (2015) A novel role of Rab11 in trafficking GPI-anchored trans-sialidase to the plasma membrane of Trypanosoma cruzi. Small GTPases 6:8-10
Jimenez, Veronica; Docampo, Roberto (2015) TcPho91 is a contractile vacuole phosphate sodium symporter that regulates phosphate and polyphosphate metabolism in Trypanosoma cruzi. Mol Microbiol 97:911-25
Ulrich, Paul N; Lander, Noelia; Kurup, Samarchith P et al. (2014) The acidocalcisome vacuolar transporter chaperone 4 catalyzes the synthesis of polyphosphate in insect-stages of Trypanosoma brucei and T. cruzi. J Eukaryot Microbiol 61:155-65

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