<named-content content-type="genus-species">Plasmodium falciparum</named-content> Resistance to a Lead Benzoxaborole Due to Blocked Compound Activation and Altered Ubiquitination or Sumoylation

ABSTRACT New antimalarial drugs are needed. The benzoxaborole AN13762 showed excellent activity against cultured Plasmodium falciparum, against fresh Ugandan P. falciparum isolates, and in murine malaria models. To gain mechanistic insights, we selected in vitro for P. falciparum isolates resistant...

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Autores principales:	Kirthana M. V. Sindhe, Wesley Wu, Jenny Legac, Yong-Kang Zhang, Eric E. Easom, Roland A. Cooper, Jacob J. Plattner, Yvonne R. Freund, Joseph L. DeRisi, Philip J. Rosenthal
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	malaria Plasmodium falciparum drug benzoxaborole resistance PfPARE Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/f9067457c1d346e893d1e74840fb86f1
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spelling	oai:doaj.org-article:f9067457c1d346e893d1e74840fb86f12021-11-15T15:56:57Z<named-content content-type="genus-species">Plasmodium falciparum</named-content> Resistance to a Lead Benzoxaborole Due to Blocked Compound Activation and Altered Ubiquitination or Sumoylation10.1128/mBio.02640-192150-7511https://doaj.org/article/f9067457c1d346e893d1e74840fb86f12020-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02640-19https://doaj.org/toc/2150-7511ABSTRACT New antimalarial drugs are needed. The benzoxaborole AN13762 showed excellent activity against cultured Plasmodium falciparum, against fresh Ugandan P. falciparum isolates, and in murine malaria models. To gain mechanistic insights, we selected in vitro for P. falciparum isolates resistant to AN13762. In all of 11 independent selections with 100 to 200 nM AN13762, the 50% inhibitory concentration (IC50) increased from 18–118 nM to 180–890 nM, and whole-genome sequencing of resistant parasites demonstrated mutations in prodrug activation and resistance esterase (PfPARE). The introduction of PfPARE mutations led to a similar level of resistance, and recombinant PfPARE hydrolyzed AN13762 to the benzoxaborole AN10248, which has activity similar to that of AN13762 but for which selection of resistance was not readily achieved. Parasites further selected with micromolar concentrations of AN13762 developed higher-level resistance (IC50, 1.9 to 5.0 μM), and sequencing revealed additional mutations in any of 5 genes, 4 of which were associated with ubiquitination/sumoylation enzyme cascades; the introduction of one of these mutations, in SUMO-activating enzyme subunit 2, led to a similar level of resistance. The other gene mutated in highly resistant parasites encodes the P. falciparum cleavage and specificity factor homolog PfCPSF3, previously identified as the antimalarial target of another benzoxaborole. Parasites selected for resistance to AN13762 were cross-resistant with a close analog, AN13956, but not with standard antimalarials, AN10248, or other benzoxaboroles known to have different P. falciparum targets. Thus, AN13762 appears to have a novel mechanism of antimalarial action and multiple mechanisms of resistance, including loss of function of PfPARE preventing activation to AN10248, followed by alterations in ubiquitination/sumoylation pathways or PfCPSF3. IMPORTANCE Benzoxaboroles are under study as potential new drugs to treat malaria. One benzoxaborole, AN13762, has potent activity and promising features, but its mechanisms of action and resistance are unknown. To gain insights into these mechanisms, we cultured malaria parasites with nonlethal concentrations of AN13762 and generated parasites with varied levels of resistance. Parasites with low-level resistance had mutations in PfPARE, which processes AN13762 into an active metabolite; PfPARE mutations prevented this processing. Parasites with high-level resistance had mutations in any of a number of enzymes, mostly those involved in stress responses. Parasites selected for AN13762 resistance were not resistant to other antimalarials, suggesting novel mechanisms of action and resistance for AN13762, a valuable feature for a new class of antimalarial drugs.Kirthana M. V. SindheWesley WuJenny LegacYong-Kang ZhangEric E. EasomRoland A. CooperJacob J. PlattnerYvonne R. FreundJoseph L. DeRisiPhilip J. RosenthalAmerican Society for MicrobiologyarticlemalariaPlasmodium falciparumdrugbenzoxaboroleresistancePfPAREMicrobiologyQR1-502ENmBio, Vol 11, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	malaria Plasmodium falciparum drug benzoxaborole resistance PfPARE Microbiology QR1-502
spellingShingle	malaria Plasmodium falciparum drug benzoxaborole resistance PfPARE Microbiology QR1-502 Kirthana M. V. Sindhe Wesley Wu Jenny Legac Yong-Kang Zhang Eric E. Easom Roland A. Cooper Jacob J. Plattner Yvonne R. Freund Joseph L. DeRisi Philip J. Rosenthal <named-content content-type="genus-species">Plasmodium falciparum</named-content> Resistance to a Lead Benzoxaborole Due to Blocked Compound Activation and Altered Ubiquitination or Sumoylation
description	ABSTRACT New antimalarial drugs are needed. The benzoxaborole AN13762 showed excellent activity against cultured Plasmodium falciparum, against fresh Ugandan P. falciparum isolates, and in murine malaria models. To gain mechanistic insights, we selected in vitro for P. falciparum isolates resistant to AN13762. In all of 11 independent selections with 100 to 200 nM AN13762, the 50% inhibitory concentration (IC50) increased from 18–118 nM to 180–890 nM, and whole-genome sequencing of resistant parasites demonstrated mutations in prodrug activation and resistance esterase (PfPARE). The introduction of PfPARE mutations led to a similar level of resistance, and recombinant PfPARE hydrolyzed AN13762 to the benzoxaborole AN10248, which has activity similar to that of AN13762 but for which selection of resistance was not readily achieved. Parasites further selected with micromolar concentrations of AN13762 developed higher-level resistance (IC50, 1.9 to 5.0 μM), and sequencing revealed additional mutations in any of 5 genes, 4 of which were associated with ubiquitination/sumoylation enzyme cascades; the introduction of one of these mutations, in SUMO-activating enzyme subunit 2, led to a similar level of resistance. The other gene mutated in highly resistant parasites encodes the P. falciparum cleavage and specificity factor homolog PfCPSF3, previously identified as the antimalarial target of another benzoxaborole. Parasites selected for resistance to AN13762 were cross-resistant with a close analog, AN13956, but not with standard antimalarials, AN10248, or other benzoxaboroles known to have different P. falciparum targets. Thus, AN13762 appears to have a novel mechanism of antimalarial action and multiple mechanisms of resistance, including loss of function of PfPARE preventing activation to AN10248, followed by alterations in ubiquitination/sumoylation pathways or PfCPSF3. IMPORTANCE Benzoxaboroles are under study as potential new drugs to treat malaria. One benzoxaborole, AN13762, has potent activity and promising features, but its mechanisms of action and resistance are unknown. To gain insights into these mechanisms, we cultured malaria parasites with nonlethal concentrations of AN13762 and generated parasites with varied levels of resistance. Parasites with low-level resistance had mutations in PfPARE, which processes AN13762 into an active metabolite; PfPARE mutations prevented this processing. Parasites with high-level resistance had mutations in any of a number of enzymes, mostly those involved in stress responses. Parasites selected for AN13762 resistance were not resistant to other antimalarials, suggesting novel mechanisms of action and resistance for AN13762, a valuable feature for a new class of antimalarial drugs.
format	article
author	Kirthana M. V. Sindhe Wesley Wu Jenny Legac Yong-Kang Zhang Eric E. Easom Roland A. Cooper Jacob J. Plattner Yvonne R. Freund Joseph L. DeRisi Philip J. Rosenthal
author_facet	Kirthana M. V. Sindhe Wesley Wu Jenny Legac Yong-Kang Zhang Eric E. Easom Roland A. Cooper Jacob J. Plattner Yvonne R. Freund Joseph L. DeRisi Philip J. Rosenthal
author_sort	Kirthana M. V. Sindhe
title	<named-content content-type="genus-species">Plasmodium falciparum</named-content> Resistance to a Lead Benzoxaborole Due to Blocked Compound Activation and Altered Ubiquitination or Sumoylation
title_short	<named-content content-type="genus-species">Plasmodium falciparum</named-content> Resistance to a Lead Benzoxaborole Due to Blocked Compound Activation and Altered Ubiquitination or Sumoylation
title_full	<named-content content-type="genus-species">Plasmodium falciparum</named-content> Resistance to a Lead Benzoxaborole Due to Blocked Compound Activation and Altered Ubiquitination or Sumoylation
title_fullStr	<named-content content-type="genus-species">Plasmodium falciparum</named-content> Resistance to a Lead Benzoxaborole Due to Blocked Compound Activation and Altered Ubiquitination or Sumoylation
title_full_unstemmed	<named-content content-type="genus-species">Plasmodium falciparum</named-content> Resistance to a Lead Benzoxaborole Due to Blocked Compound Activation and Altered Ubiquitination or Sumoylation
title_sort	<named-content content-type="genus-species">plasmodium falciparum</named-content> resistance to a lead benzoxaborole due to blocked compound activation and altered ubiquitination or sumoylation
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/f9067457c1d346e893d1e74840fb86f1
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<named-content content-type="genus-species">Plasmodium falciparum</named-content> Resistance to a Lead Benzoxaborole Due to Blocked Compound Activation and Altered Ubiquitination or Sumoylation

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