Identification and Mechanistic Understanding of Dihydroorotate Dehydrogenase Point Mutations in Plasmodium falciparum that Confer in Vitro Resistance to the Clinical Candidate DSM265

John White, Satish K. Dhingra, Xiaoyi Deng, Farah El Mazouni, Marcus C.S. Lee, Gustavo A. Afanador, Aloysus Lawong, Diana R Tomchick, Caroline L. Ng, Jade Bath, Pradipsinh K. Rathod, David A. Fidock, Margaret A Phillips

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Malaria is one of the most challenging human infectious diseases, and both prevention and control have been hindered by the development of Plasmodium falciparum resistance to existing therapies. Several new compounds with novel mechanisms are in clinical development for the treatment of malaria, including DSM265, an inhibitor of Plasmodium dihydroorotate dehydrogenase. To explore the mechanisms by which resistance might develop to DSM265 in the field, we selected for DSM265-resistant P. falciparum parasites in vitro. Any of five different amino acid changes led to reduced efficacy on the parasite and to decreased DSM265 binding to P. falciparum DHODH. The DSM265-resistant parasites retained full sensitivity to atovaquone. All but one of the observed mutations were in the DSM265 binding site, and the remaining C276F was in the adjacent flavin cofactor site. The C276F mutation was previously identified in a recrudescent parasite during a Phase IIa clinical study. We confirmed that this mutation (and the related C276Y) accounted for the full level of observed DSM265 resistance by regenerating the mutation using CRISPR/Cas9 genome editing. X-ray structure analysis of the C276F mutant enzyme showed that conformational changes of nearby residues were required to accommodate the larger F276 residue, which in turn led to a restriction in the size of the DSM265 binding pocket. These findings underscore the importance of developing DSM265 as part of a combination therapy with other agents for successful use against malaria.

Original languageEnglish (US)
JournalACS Infectious Diseases
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Plasmodium falciparum
Point Mutation
Parasites
Malaria
Mutation
Clustered Regularly Interspaced Short Palindromic Repeats
Atovaquone
DSM265
In Vitro Techniques
dihydroorotate dehydrogenase
Plasmodium
Communicable Diseases
Binding Sites
X-Rays
Amino Acids
Enzymes
Therapeutics

Keywords

  • dihydroorotate dehydrogenase
  • drug resistance
  • gene editing
  • malaria
  • Plasmodium

ASJC Scopus subject areas

  • Infectious Diseases

Cite this

Identification and Mechanistic Understanding of Dihydroorotate Dehydrogenase Point Mutations in Plasmodium falciparum that Confer in Vitro Resistance to the Clinical Candidate DSM265. / White, John; Dhingra, Satish K.; Deng, Xiaoyi; El Mazouni, Farah; Lee, Marcus C.S.; Afanador, Gustavo A.; Lawong, Aloysus; Tomchick, Diana R; Ng, Caroline L.; Bath, Jade; Rathod, Pradipsinh K.; Fidock, David A.; Phillips, Margaret A.

In: ACS Infectious Diseases, 01.01.2018.

Research output: Contribution to journalArticle

White, John ; Dhingra, Satish K. ; Deng, Xiaoyi ; El Mazouni, Farah ; Lee, Marcus C.S. ; Afanador, Gustavo A. ; Lawong, Aloysus ; Tomchick, Diana R ; Ng, Caroline L. ; Bath, Jade ; Rathod, Pradipsinh K. ; Fidock, David A. ; Phillips, Margaret A. / Identification and Mechanistic Understanding of Dihydroorotate Dehydrogenase Point Mutations in Plasmodium falciparum that Confer in Vitro Resistance to the Clinical Candidate DSM265. In: ACS Infectious Diseases. 2018.
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