Interaction networks of intracellular pathogens and their host cells are complex and predicted to be adaptive in promoting pathogen survival. During malaria parasite intrahepatocytic liver stage infection, parasites protect their host hepatocyte by preventing its death and fully exploit the host cell resources for growth and replication. The host hepatocyte molecular signaling landscape that undergirds successful liver stage replication has not been elucidated, yet it is highly medically relevant. We will use wild-type and attenuated parasite strains that arrest at different points after hepatocyte infection as a tool to investigat the differences between how healthy, wild-type parasites manipulate their host cell and how attenuated parasites, which are cleared from the liver shortly after arrest, are unable to do so. I this proposal, we propose experiments that fully delineate the pro- apoptotic milieu that the parasite can evade, and what intrinsic and extrinsic perturbations lead to the demise of even a wild-type parasite. Next, we will expand upon our already intriguing molecular dataset by extensively interrogating signaling proteins in hepatocytes in response to both wild-type and attenuated parasites using protein lysate microarrays, an approach that allows us to monitor hundreds of protein and post-translational modification levels using lysates derived from ~10,000 liver stage-infected hepatocytes. We propose monitoring hepatocyte signals not only in response to rodent malaria infection, but also in response to the most deadly human malaria parasite, Plasmodium falciparum, using infected hepatocytes from a mouse with a humanized liver. In preliminary studies, we have recently correlated the down-regulation of the tumor suppressor gene p53 with successful Plasmodium liver stage infection, and have demonstrated that artificially increasing p53 levels can eliminate liver stage parasites. We plan to further elucidate the mechanism behind the parasite's dependence on low host p53 for survival. Finally, we investigate the role that BH3-domain containing proteins in the mitochondria of hepatocytes play in malaria parasite liver-stage infection and ask if they are targeted by the parasite to prevent host cell apoptosis. The proposed studies will lead to a more comprehensive understanding of the hepatocyte signaling landscape critical to modulating the host response to Plasmodium liver stage parasite infection, including unparalleled understanding of parasites that infect humans and that impact human health. Accomplishing our aims opens the possibility of altering the hepatocyte signaling landscape with small-molecules that could prevent a wild-type parasite from progressing to symptomatic erythrocyte infection. Such a host-based approach for prophylaxis is novel and will circumvent the massive problem of continuously developing resistance to standard antimalarial drugs. This approach is further fostered by the fact that many hepatocyte proteins are already targets of known therapeutic inhibitors. Perturbations of hepatocyte signaling by a complex intracellular pathogen might also reveal new intrinsic features of the signaling system within hepatocytes.

Public Health Relevance

The complex web of signals that arise from malaria parasite development within a host hepatocyte is highly medically relevant, as malaria kills more than one million people annually. This proposal seeks to investigate the molecular signals by which the hepatocyte and intracellular pathogen interact over the course of the parasite's liver stage development.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM101183-03
Application #
8827809
Study Section
Cellular Signaling and Regulatory Systems Study Section (CSRS)
Program Officer
Sledjeski, Darren D
Project Start
2013-04-01
Project End
2017-03-31
Budget Start
2015-04-01
Budget End
2016-03-31
Support Year
3
Fiscal Year
2015
Total Cost
$359,100
Indirect Cost
$169,100
Name
Seattle Biomedical Research Institute
Department
Type
DUNS #
070967955
City
Seattle
State
WA
Country
United States
Zip Code
98109
Patterson, Nathan Heath; Tuck, Michael; Lewis, Adam et al. (2018) Next Generation Histology-Directed Imaging Mass Spectrometry Driven by Autofluorescence Microscopy. Anal Chem 90:12404-12413
Glennon, Elizabeth K K; Dankwa, Selasi; Smith, Joseph D et al. (2018) Opportunities for Host-targeted Therapies for Malaria. Trends Parasitol 34:843-860
Zuck, Meghan; Austin, Laura S; Danziger, Samuel A et al. (2017) The Promise of Systems Biology Approaches for Revealing Host Pathogen Interactions in Malaria. Front Microbiol 8:2183
Arang, Nadia; Kain, Heather S; Glennon, Elizabeth K et al. (2017) Identifying host regulators and inhibitors of liver stage malaria infection using kinase activity profiles. Nat Commun 8:1232
Kaushansky, Alexis; Kappe, Stefan Hi (2015) Selection and refinement: the malaria parasite's infection and exploitation of host hepatocytes. Curr Opin Microbiol 26:71-8
Douglass, Alyse N; Kain, Heather S; Abdullahi, Marian et al. (2015) Host-based Prophylaxis Successfully Targets Liver Stage Malaria Parasites. Mol Ther 23:857-865
Kaushansky, Alexis; Douglass, Alyse N; Arang, Nadia et al. (2015) Malaria parasites target the hepatocyte receptor EphA2 for successful host infection. Science 350:1089-92
Kaushansky, Alexis; Austin, Laura S; Mikolajczak, Sebastian A et al. (2015) Susceptibility to Plasmodium yoelii preerythrocytic infection in BALB/c substrains is determined at the point of hepatocyte invasion. Infect Immun 83:39-47
Austin, Laura S; Kaushansky, Alexis; Kappe, Stefan H I (2014) Susceptibility to Plasmodium liver stage infection is altered by hepatocyte polyploidy. Cell Microbiol 16:784-95
Kaushansky, A; Metzger, P G; Douglass, A N et al. (2013) Malaria parasite liver stages render host hepatocytes susceptible to mitochondria-initiated apoptosis. Cell Death Dis 4:e762