Assessing drug efficacy against Plasmodium falciparum liver stages in vivo

Erika L. Flannery, Lander Foquet, Vorada Chuenchob, Matthew Fishbaugher, Zachary Billman, Mary Jane Navarro, William Betz, Tayla M. Olsen, Joshua Lee, Nelly Camargo, Thao Nguyen, Carola Schafer, Brandon K. Sack, Elizabeth M. Wilson, Jessica Saunders, John Bial, Brice Campo, Susan A. Charman, Sean C. Murphy, Margaret A. Phillips & 2 others Stefan Hi Kappe, Sebastian A. Mikolajczak

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Malaria eradication necessitates new tools to fight the evolving and complex Plasmodium pathogens. These tools include prophylactic drugs that eliminate Plasmodium liver stages and consequently prevent clinical disease, decrease transmission, and reduce the propensity for resistance development. Currently, the identification of these drugs relies on in vitro P. falciparum liver stage assays or in vivo causal prophylaxis assays using rodent malaria parasites; there is no method to directly test in vivo liver stage activity of candidate antimalarials against the human malaria-causing parasite P. falciparum. Here, we use a liver-chimeric humanized mouse (FRG huHep) to demonstrate in vivo P. falciparum liver stage development and describe the efficacy of clinically used and candidate antimalarials with prophylactic activity. We show that daily administration of atovaquone-proguanil (ATQ-PG; ATQ, 30 mg/kg, and PG, 10 mg/kg) protects 5 of 5 mice from liver stage infection, consistent with the use in humans as a causal prophylactic drug. Single-dose primaquine (60 mg/kg) has similar activity to that observed in humans, demonstrating the activity of this drug (and its active metabolites) in FRG huHep mice. We also show that DSM265, a selective Plasmodial dihydroorotate dehydrogenase inhibitor with causal prophylactic activity in humans, reduces liver stage burden in FRG huHep mice. Finally, we measured liver stage-to-blood stage transition of the parasite, the ultimate readout of prophylactic activity and measurement of infective capacity of parasites in the liver, to show that ATQ-PG reduces blood stage patency to below the limit of quantitation by quantitative PCR (qPCR). The FRG huHep model, thus, provides a platform for preclinical evaluation of drug candidates for liver stage causal prophylactic activity, pharmacokinetic/pharmacodynamics studies, and biological studies to investigate the mechanism of action of liver stage active antimalarials.

Original languageEnglish
Article numbere92587
Number of pages12
JournalJCI Insight
Volume3
Issue number1
DOIs
Publication statusPublished - 11 Jan 2018

Keywords

  • Drug screens
  • Infectious disease
  • Malaria
  • Microbiology
  • Mouse models

Cite this

Flannery, E. L., Foquet, L., Chuenchob, V., Fishbaugher, M., Billman, Z., Navarro, M. J., ... Mikolajczak, S. A. (2018). Assessing drug efficacy against Plasmodium falciparum liver stages in vivo. JCI Insight, 3(1), [e92587]. https://doi.org/10.1172/jci.insight.92587
Flannery, Erika L. ; Foquet, Lander ; Chuenchob, Vorada ; Fishbaugher, Matthew ; Billman, Zachary ; Navarro, Mary Jane ; Betz, William ; Olsen, Tayla M. ; Lee, Joshua ; Camargo, Nelly ; Nguyen, Thao ; Schafer, Carola ; Sack, Brandon K. ; Wilson, Elizabeth M. ; Saunders, Jessica ; Bial, John ; Campo, Brice ; Charman, Susan A. ; Murphy, Sean C. ; Phillips, Margaret A. ; Kappe, Stefan Hi ; Mikolajczak, Sebastian A. / Assessing drug efficacy against Plasmodium falciparum liver stages in vivo. In: JCI Insight. 2018 ; Vol. 3, No. 1.
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abstract = "Malaria eradication necessitates new tools to fight the evolving and complex Plasmodium pathogens. These tools include prophylactic drugs that eliminate Plasmodium liver stages and consequently prevent clinical disease, decrease transmission, and reduce the propensity for resistance development. Currently, the identification of these drugs relies on in vitro P. falciparum liver stage assays or in vivo causal prophylaxis assays using rodent malaria parasites; there is no method to directly test in vivo liver stage activity of candidate antimalarials against the human malaria-causing parasite P. falciparum. Here, we use a liver-chimeric humanized mouse (FRG huHep) to demonstrate in vivo P. falciparum liver stage development and describe the efficacy of clinically used and candidate antimalarials with prophylactic activity. We show that daily administration of atovaquone-proguanil (ATQ-PG; ATQ, 30 mg/kg, and PG, 10 mg/kg) protects 5 of 5 mice from liver stage infection, consistent with the use in humans as a causal prophylactic drug. Single-dose primaquine (60 mg/kg) has similar activity to that observed in humans, demonstrating the activity of this drug (and its active metabolites) in FRG huHep mice. We also show that DSM265, a selective Plasmodial dihydroorotate dehydrogenase inhibitor with causal prophylactic activity in humans, reduces liver stage burden in FRG huHep mice. Finally, we measured liver stage-to-blood stage transition of the parasite, the ultimate readout of prophylactic activity and measurement of infective capacity of parasites in the liver, to show that ATQ-PG reduces blood stage patency to below the limit of quantitation by quantitative PCR (qPCR). The FRG huHep model, thus, provides a platform for preclinical evaluation of drug candidates for liver stage causal prophylactic activity, pharmacokinetic/pharmacodynamics studies, and biological studies to investigate the mechanism of action of liver stage active antimalarials.",
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Flannery, EL, Foquet, L, Chuenchob, V, Fishbaugher, M, Billman, Z, Navarro, MJ, Betz, W, Olsen, TM, Lee, J, Camargo, N, Nguyen, T, Schafer, C, Sack, BK, Wilson, EM, Saunders, J, Bial, J, Campo, B, Charman, SA, Murphy, SC, Phillips, MA, Kappe, SH & Mikolajczak, SA 2018, 'Assessing drug efficacy against Plasmodium falciparum liver stages in vivo' JCI Insight, vol. 3, no. 1, e92587. https://doi.org/10.1172/jci.insight.92587

Assessing drug efficacy against Plasmodium falciparum liver stages in vivo. / Flannery, Erika L.; Foquet, Lander; Chuenchob, Vorada; Fishbaugher, Matthew; Billman, Zachary; Navarro, Mary Jane; Betz, William; Olsen, Tayla M.; Lee, Joshua; Camargo, Nelly; Nguyen, Thao; Schafer, Carola; Sack, Brandon K.; Wilson, Elizabeth M.; Saunders, Jessica; Bial, John; Campo, Brice; Charman, Susan A.; Murphy, Sean C.; Phillips, Margaret A.; Kappe, Stefan Hi; Mikolajczak, Sebastian A.

In: JCI Insight, Vol. 3, No. 1, e92587, 11.01.2018.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Assessing drug efficacy against Plasmodium falciparum liver stages in vivo

AU - Flannery, Erika L.

AU - Foquet, Lander

AU - Chuenchob, Vorada

AU - Fishbaugher, Matthew

AU - Billman, Zachary

AU - Navarro, Mary Jane

AU - Betz, William

AU - Olsen, Tayla M.

AU - Lee, Joshua

AU - Camargo, Nelly

AU - Nguyen, Thao

AU - Schafer, Carola

AU - Sack, Brandon K.

AU - Wilson, Elizabeth M.

AU - Saunders, Jessica

AU - Bial, John

AU - Campo, Brice

AU - Charman, Susan A.

AU - Murphy, Sean C.

AU - Phillips, Margaret A.

AU - Kappe, Stefan Hi

AU - Mikolajczak, Sebastian A.

PY - 2018/1/11

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N2 - Malaria eradication necessitates new tools to fight the evolving and complex Plasmodium pathogens. These tools include prophylactic drugs that eliminate Plasmodium liver stages and consequently prevent clinical disease, decrease transmission, and reduce the propensity for resistance development. Currently, the identification of these drugs relies on in vitro P. falciparum liver stage assays or in vivo causal prophylaxis assays using rodent malaria parasites; there is no method to directly test in vivo liver stage activity of candidate antimalarials against the human malaria-causing parasite P. falciparum. Here, we use a liver-chimeric humanized mouse (FRG huHep) to demonstrate in vivo P. falciparum liver stage development and describe the efficacy of clinically used and candidate antimalarials with prophylactic activity. We show that daily administration of atovaquone-proguanil (ATQ-PG; ATQ, 30 mg/kg, and PG, 10 mg/kg) protects 5 of 5 mice from liver stage infection, consistent with the use in humans as a causal prophylactic drug. Single-dose primaquine (60 mg/kg) has similar activity to that observed in humans, demonstrating the activity of this drug (and its active metabolites) in FRG huHep mice. We also show that DSM265, a selective Plasmodial dihydroorotate dehydrogenase inhibitor with causal prophylactic activity in humans, reduces liver stage burden in FRG huHep mice. Finally, we measured liver stage-to-blood stage transition of the parasite, the ultimate readout of prophylactic activity and measurement of infective capacity of parasites in the liver, to show that ATQ-PG reduces blood stage patency to below the limit of quantitation by quantitative PCR (qPCR). The FRG huHep model, thus, provides a platform for preclinical evaluation of drug candidates for liver stage causal prophylactic activity, pharmacokinetic/pharmacodynamics studies, and biological studies to investigate the mechanism of action of liver stage active antimalarials.

AB - Malaria eradication necessitates new tools to fight the evolving and complex Plasmodium pathogens. These tools include prophylactic drugs that eliminate Plasmodium liver stages and consequently prevent clinical disease, decrease transmission, and reduce the propensity for resistance development. Currently, the identification of these drugs relies on in vitro P. falciparum liver stage assays or in vivo causal prophylaxis assays using rodent malaria parasites; there is no method to directly test in vivo liver stage activity of candidate antimalarials against the human malaria-causing parasite P. falciparum. Here, we use a liver-chimeric humanized mouse (FRG huHep) to demonstrate in vivo P. falciparum liver stage development and describe the efficacy of clinically used and candidate antimalarials with prophylactic activity. We show that daily administration of atovaquone-proguanil (ATQ-PG; ATQ, 30 mg/kg, and PG, 10 mg/kg) protects 5 of 5 mice from liver stage infection, consistent with the use in humans as a causal prophylactic drug. Single-dose primaquine (60 mg/kg) has similar activity to that observed in humans, demonstrating the activity of this drug (and its active metabolites) in FRG huHep mice. We also show that DSM265, a selective Plasmodial dihydroorotate dehydrogenase inhibitor with causal prophylactic activity in humans, reduces liver stage burden in FRG huHep mice. Finally, we measured liver stage-to-blood stage transition of the parasite, the ultimate readout of prophylactic activity and measurement of infective capacity of parasites in the liver, to show that ATQ-PG reduces blood stage patency to below the limit of quantitation by quantitative PCR (qPCR). The FRG huHep model, thus, provides a platform for preclinical evaluation of drug candidates for liver stage causal prophylactic activity, pharmacokinetic/pharmacodynamics studies, and biological studies to investigate the mechanism of action of liver stage active antimalarials.

KW - Drug screens

KW - Infectious disease

KW - Malaria

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KW - Mouse models

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Flannery EL, Foquet L, Chuenchob V, Fishbaugher M, Billman Z, Navarro MJ et al. Assessing drug efficacy against Plasmodium falciparum liver stages in vivo. JCI Insight. 2018 Jan 11;3(1). e92587. https://doi.org/10.1172/jci.insight.92587