Unraveling the biology of human pathogens is fundamental toward understanding mechanisms of pathogenesis and identifying genes essential for survival in the host. This application focuses on the protozoan parasite Trypanosoma brucei, which causes devastating diseases in humans and animals in sub-Saharan Africa. There are no vaccines, and therapeutic drugs have serious side effects and decreasing efficacy. T. brucei undergoes a complex life cycle between the mammalian host and the blood-feeding tsetse fly vector (Diptera: Glossinidae), which among others involves changes in cell morphology, metabolism, signaling pathways and gene expression. Consequently, these parasites have evolved adaptations to allow for their survival in both the gut and salivary glands of the tsetse fly, as well as in the bloodstream of their mammalian host. Upon feeding on an infected host, the tsetse fly takes up slender, intermediate and stumpy bloodstream forms. In the fly midgut stumpy forms differentiate into non-infectious procyclic forms. Reacquisition of infectivity is achieved through a complex developmental program that culminates in the tsetse salivary glands with the generation of infectious metacyclics. Although the intricate nature of trypanosome development in the fly has been recognized for more than a century, the molecular mechanisms are still mysterious, due in part to the experimental challenges posed by the tsetse fly. We found that overexpression of the T. brucei RNA-binding protein RBP6 in cultured non-infectious procyclic forms initiates differentiation into the developmental stages found in tsetse flies and culminates with the generation of infective metacyclics expressing the variant surface glycoprotein (VSG) coat. Our first goal will be to delineate the mechanism of action of RBP6. As RBP6 appears to be a "master regulator" triggering a cascade of events, it will be critical to identify the primary mRNA targets of RBP6. This will provide crucial information about gene products involved in the early stages of differentiation and for formulating a testable hypothesis about the cellular adaptations occurring in the transition from procyclic to epimastigotes forms. Our second goal will be to provide a transcriptomic and proteomic map of epimastigotes and metacyclics and to decode the biology of metacyclogenesis. Finally, the discovery of novel genes, besides VSG, required for metacyclic differentiation will be a major breakthrough toward deciphering how the process of acquisition of infectivity is brought about. Taken together our research plan provides unique opportunities to illuminate the differentiation program from procyclic to metacyclic and reveal the mode of action of an important RNA binding protein.

Public Health Relevance

Parasitic protozoa are a major cause of global infectious diseases and thus, represent one of the most serious threats to public health. Among these are African trypanosomes, the causative agents of African trypanosomiasis or sleeping sickness in humans and a wasting and fatal disease (Nagana) in cattle, domestic pigs and other farm animals causing a profound effect on the economy of much of the continent. Our proposed studies will provide an insight into the developmental program that leads to the establishment of infectious trypanosomes, which has the potential to expose new strategies to combat the diseases caused by these deadly parasites.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI110325-01
Application #
8660833
Study Section
Special Emphasis Panel (ZRG1-IDM-M (02))
Program Officer
Mcgugan, Glen C
Project Start
2014-03-15
Project End
2019-02-28
Budget Start
2014-03-15
Budget End
2015-02-28
Support Year
1
Fiscal Year
2014
Total Cost
$416,250
Indirect Cost
$166,250
Name
Yale University
Department
Public Health & Prev Medicine
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520