The mitochondrion of P. falciparum is stripped of many of the typical functions of mitochondria, yet retains pathways essential to the parasite survival. The remaining mitochondrial functions are proving to be an exceptionally productive set of targets for antimalarial drug discovery. One approach to overcoming drug resistance is to design new drugs such that the fitness cost of resistance restricts the ability of mutant parasites to survive combination therapy or to persist in the absence of selection. Through whole organism high-throughput screening we have discovered many new chemotypes that appear to act on mitochondrial targets. We propose to take a chemical biology approach to the mitochondrion and develop these into a suite of reagents useful in characterizing mitochondrial function by pinpointing mechanisms of action and resistance for a range of chemotypes that act against the mitochondrion. We will study the effects of the resistance mutations on the structure and function of the targets and the impact of those mutations and consequent biochemical changes on parasite growth and fitness in vitro. We will select resistance to a range of chemical inhibitors that appear to target the mitochondrial ETC, focusing on DHODH. Using an approach that has proven highly productive, we will characterize the resistant mutants to identify the target of the chemical inhibitor. We propose to conduct selections in sufficient depth and using a sufficiently diverse range of chemistry to sample the range of possible targets and mechanisms affecting the mitochondrial ETC, and to explore the range of resistance mutations to DHODH inhibitors intensively, focusing on the compound advancing to clinical testing. We will screen previously identified DHODH inhibitor screening hits for those active against the resistant parasites. We will map the resistance mutations of mutants newly isolated and assess the biological consequences by measuring the effects of the mutations on the enzyme, on mitochondrial function and on the growth of the organism.

Public Health Relevance

In this project we will develop chemical and genetic probes to explore the role and function of the mitochondrion of the malaria parasite. We will find new anti- malarial compounds that target mitochondrial functions and then select mutations resistant to those compounds to pinpoint the exact target within the mitochondrion. Often the mutations that give rise to resistance hamper the ability of the parasite to develop and grow normally;we will study the effect of the mutations on growth to allow us to better choose new drug candidates or combinations of drugs less likely to select resistance when the drugs are given to patients.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI093716-02
Application #
8505368
Study Section
Drug Discovery and Mechanisms of Antimicrobial Resistance Study Section (DDR)
Program Officer
Rogers, Martin J
Project Start
2012-07-06
Project End
2016-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
2
Fiscal Year
2013
Total Cost
$423,041
Indirect Cost
$111,078
Name
Broad Institute, Inc.
Department
Type
DUNS #
623544785
City
Cambridge
State
MA
Country
United States
Zip Code
02142
Ross, Leila S; Gamo, Francisco Javier; Lafuente-Monasterio, Maria José et al. (2014) In vitro resistance selections for Plasmodium falciparum dihydroorotate dehydrogenase inhibitors give mutants with multiple point mutations in the drug-binding site and altered growth. J Biol Chem 289:17980-95
Comer, Eamon; Beaudoin, Jennifer A; Kato, Nobutaka et al. (2014) Diversity-oriented synthesis-facilitated medicinal chemistry: toward the development of novel antimalarial agents. J Med Chem 57:8496-502
Lukens, Amanda K; Ross, Leila Saxby; Heidebrecht, Richard et al. (2014) Harnessing evolutionary fitness in Plasmodium falciparum for drug discovery and suppressing resistance. Proc Natl Acad Sci U S A 111:799-804