Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility

Detalhes bibliográficos
Autor(a) principal: Santi, Ana Maria Murta
Data de Publicação: 2021
Outros Autores: Silva, Paula Alves, Santos, Isabella Fernandes Martins, Murta, Silvane Maria Fonseca
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/51547
Resumo: Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Genômica Funcional de Parasitos. Belo Horizonte, MG, Brazil.
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spelling Santi, Ana Maria MurtaSilva, Paula AlvesSantos, Isabella Fernandes MartinsMurta, Silvane Maria Fonseca2022-03-04T13:31:09Z2022-03-04T13:31:09Z2021SANTI, Ana Maria Murta et al. Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility. Parasit Vectors, v. 14, n. 1, 366, 2021. doi: 10.1186/s13071-021-04838-8.1756-3305https://www.arca.fiocruz.br/handle/icict/5154710.1186/s13071-021-04838-8.engBioMed CentralDownregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibilityinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto René Rachou. Grupo de Genômica Funcional de Parasitos. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Genômica Funcional de Parasitos. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Genômica Funcional de Parasitos. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Genômica Funcional de Parasitos. Belo Horizonte, MG, Brazil.Background: Superoxide dismutase (SOD), a central component of the antioxidant defence system of most organisms, removes excess superoxide anions by converting them to oxygen and hydrogen peroxide. As iron (Fe) SOD is absent in the human host, this enzyme is a promising molecular target for drug development against trypanosomatids. Results: We obtained Leishmania infantum mutant clones with lower FeSOD-A expression and investigated their phenotypes. Our attempts to delete this enzyme-coding gene using three different methodologies (conventional allelic replacement or two different CRISPR/methods) failed, as FeSOD-A gene copies were probably retained by aneuploidy or gene amplification. Promastigote forms of WT and mutant parasites were used in quantitative reverse-transcription polymerase chain reaction (RT-qPCR) and western blot analyses, and these parasite forms were also used to assess drug susceptibility. RT-qPCR and western blot analyses revealed that FeSOD-A transcript and protein levels were lower in FeSOD-A-/-/+ L. infantum mutant clones than in the wild-type (WT) parasite. The decrease in FeSOD-A expression in L. infantum did not interfere with the parasite growth or susceptibility to amphotericin B. Surprisingly, FeSOD-A-/-/+ L. infantum mutant clones were 1.5- to 2.0-fold more resistant to trivalent antimony and 2.4- to 2.7-fold more resistant to miltefosine. To investigate whether the decrease in FeSOD-A expression was compensated by other enzymes, the transcript levels of five FeSODs and six enzymes from the antioxidant defence system were assessed by RT-qPCR. The transcript level of the enzyme ascorbate peroxidase increased in both the FeSOD-A-/-/+ mutants tested. The FeSOD-A-/-/+ mutant parasites were 1.4- to 1.75-fold less tolerant to oxidative stress generated by menadione. Infection analysis using THP-1 macrophages showed that 72 h post-infection, the number of infected macrophages and their intracellular multiplication rate were lower in the FeSOD-A-/-/+ mutant clones than in the WT parasite. Conclusions: The unsuccessful attempts to delete FeSOD-A suggest that this gene is essential in L. infantum. This enzyme plays an important role in the defence against oxidative stress and infectivity in THP-1 macrophages. FeSOD-A-deficient L. infantum parasites deregulate their metabolic pathways related to antimony and miltefosine resistance.Drug resistanceIron superoxide dismutaseKnockoutLeishmania infantumOxidative stressinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83082https://www.arca.fiocruz.br/bitstream/icict/51547/1/license.txt9193a7c197bc67acd023525e72a03240MD51ORIGINALDownregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility.pdfDownregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility.pdfapplication/pdf5501318https://www.arca.fiocruz.br/bitstream/icict/51547/2/Downregulation%20of%20FeSOD-A%20expression%20in%20Leishmania%20infantum%20alters%20trivalent%20antimony%20and%20miltefosine%20susceptibility.pdfe2437f50c5553790caa86d9a551149d8MD52icict/515472022-03-04 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dc.title.pt_BR.fl_str_mv Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility
title Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility
spellingShingle Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility
Santi, Ana Maria Murta
Drug resistance
Iron superoxide dismutase
Knockout
Leishmania infantum
Oxidative stress
title_short Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility
title_full Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility
title_fullStr Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility
title_full_unstemmed Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility
title_sort Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility
author Santi, Ana Maria Murta
author_facet Santi, Ana Maria Murta
Silva, Paula Alves
Santos, Isabella Fernandes Martins
Murta, Silvane Maria Fonseca
author_role author
author2 Silva, Paula Alves
Santos, Isabella Fernandes Martins
Murta, Silvane Maria Fonseca
author2_role author
author
author
dc.contributor.author.fl_str_mv Santi, Ana Maria Murta
Silva, Paula Alves
Santos, Isabella Fernandes Martins
Murta, Silvane Maria Fonseca
dc.subject.en.pt_BR.fl_str_mv Drug resistance
Iron superoxide dismutase
Knockout
Leishmania infantum
Oxidative stress
topic Drug resistance
Iron superoxide dismutase
Knockout
Leishmania infantum
Oxidative stress
description Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Genômica Funcional de Parasitos. Belo Horizonte, MG, Brazil.
publishDate 2021
dc.date.issued.fl_str_mv 2021
dc.date.accessioned.fl_str_mv 2022-03-04T13:31:09Z
dc.date.available.fl_str_mv 2022-03-04T13:31:09Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv SANTI, Ana Maria Murta et al. Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility. Parasit Vectors, v. 14, n. 1, 366, 2021. doi: 10.1186/s13071-021-04838-8.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/51547
dc.identifier.issn.pt_BR.fl_str_mv 1756-3305
dc.identifier.doi.none.fl_str_mv 10.1186/s13071-021-04838-8.
identifier_str_mv SANTI, Ana Maria Murta et al. Downregulation of FeSOD-A expression in Leishmania infantum alters trivalent antimony and miltefosine susceptibility. Parasit Vectors, v. 14, n. 1, 366, 2021. doi: 10.1186/s13071-021-04838-8.
1756-3305
10.1186/s13071-021-04838-8.
url https://www.arca.fiocruz.br/handle/icict/51547
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/51547/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/51547/2/Downregulation%20of%20FeSOD-A%20expression%20in%20Leishmania%20infantum%20alters%20trivalent%20antimony%20and%20miltefosine%20susceptibility.pdf
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repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
repository.mail.fl_str_mv repositorio.arca@fiocruz.br
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