Exploring direct and indirect targets of current antileishmanial drugs using a novel thermal proteomics profiling approach
Visceral leishmaniasis (VL), caused by Leishmania infantum, is an oft-fatal neglected tropical disease. In the absence of an effective vaccine, the control of leishmaniasis relies exclusively on chemotherapy. Due to the lack of established molecular/genetic markers denoting parasite resistance, clin...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-08-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2022.954144/full |
| _version_ | 1818010957230637056 |
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| author | Ana Victoria Ibarra-Meneses Ana Victoria Ibarra-Meneses Audrey Corbeil Audrey Corbeil Victoria Wagner Victoria Wagner Francis Beaudry Francis Beaudry Rubens L. do Monte-Neto Christopher Fernandez-Prada Christopher Fernandez-Prada |
| author_facet | Ana Victoria Ibarra-Meneses Ana Victoria Ibarra-Meneses Audrey Corbeil Audrey Corbeil Victoria Wagner Victoria Wagner Francis Beaudry Francis Beaudry Rubens L. do Monte-Neto Christopher Fernandez-Prada Christopher Fernandez-Prada |
| author_sort | Ana Victoria Ibarra-Meneses |
| collection | DOAJ |
| description | Visceral leishmaniasis (VL), caused by Leishmania infantum, is an oft-fatal neglected tropical disease. In the absence of an effective vaccine, the control of leishmaniasis relies exclusively on chemotherapy. Due to the lack of established molecular/genetic markers denoting parasite resistance, clinical treatment failure is often used as an indicator. Antimony-based drugs have been the standard antileishmanial treatment for more than seven decades, leading to major drug resistance in certain regions. Likewise, drug resistance to miltefosine and amphotericin B continues to spread at alarming rates. In consequence, innovative approaches are needed to accelerate the identification of antimicrobial drug targets and resistance mechanisms. To this end, we have implemented a novel approach based on thermal proteome profiling (TPP) to further characterize the mode of action of antileishmanials antimony, miltefosine and amphotericin B, as well as to better understand the mechanisms of drug resistance deployed by Leishmania. Proteins become more resistant to heat-induced denaturation when complexed with a ligand. In this way, we used multiplexed quantitative mass spectrometry-based proteomics to monitor the melting profile of thousands of expressed soluble proteins in WT, antimony-resistant, miltefosine-resistant, and amphotericin B-resistant L. infantum parasites, in the presence (or absence) of the above-mentioned drugs. Bioinformatics analyses were performed, including data normalization, melting profile fitting, and identification of proteins that underwent changes (fold change > 4) caused by complexation with a drug. With this unique approach, we were able to narrow down the regions of the L. infantum proteome that interact with antimony, miltefosine, and amphotericin B; validating previously-identified and unveiling novel drug targets. Moreover, analyses revealed candidate proteins potentially involved in drug resistance. Interestingly, we detected thermal proximity coaggregation for several proteins belonging to the same metabolic pathway (i.e., tryparedoxin peroxidase and aspartate aminotransferase in proteins exposed to antimony), highlighting the importance of these pathways. Collectively, our results could serve as a jumping-off point for the future development of innovative diagnostic tools for the detection and evaluation of antimicrobial-resistant Leishmania populations, as well as open the door for new on-target therapies. |
| first_indexed | 2024-04-14T06:02:15Z |
| format | Article |
| id | doaj.art-fc8fe37f8be44d71943c8f1a1d4c7be3 |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-04-14T06:02:15Z |
| publishDate | 2022-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-fc8fe37f8be44d71943c8f1a1d4c7be32022-12-22T02:08:44ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882022-08-011210.3389/fcimb.2022.954144954144Exploring direct and indirect targets of current antileishmanial drugs using a novel thermal proteomics profiling approachAna Victoria Ibarra-Meneses0Ana Victoria Ibarra-Meneses1Audrey Corbeil2Audrey Corbeil3Victoria Wagner4Victoria Wagner5Francis Beaudry6Francis Beaudry7Rubens L. do Monte-Neto8Christopher Fernandez-Prada9Christopher Fernandez-Prada10Département de Pathologie et Microbiologie, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, QC, CanadaThe Research Group on Infectious Diseases in Production Animals (GREMIP), Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, QC, CanadaDépartement de Pathologie et Microbiologie, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, QC, CanadaThe Research Group on Infectious Diseases in Production Animals (GREMIP), Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, QC, CanadaDépartement de Pathologie et Microbiologie, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, QC, CanadaThe Research Group on Infectious Diseases in Production Animals (GREMIP), Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, QC, CanadaDépartement de Biomédecine, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, QC, CanadaCentre de recherche sur le cerveau et l’apprentissage (CIRCA), Université de Montréal, Montréal, QC, CanadaBiotechnology Applied to Pathogens (BAP) - Instituto René Rachou – Fundação Oswaldo Cruz/Fiocruz Minas, Belo Horizonte, Minas Gerais, BrazilDépartement de Pathologie et Microbiologie, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, QC, CanadaThe Research Group on Infectious Diseases in Production Animals (GREMIP), Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, QC, CanadaVisceral leishmaniasis (VL), caused by Leishmania infantum, is an oft-fatal neglected tropical disease. In the absence of an effective vaccine, the control of leishmaniasis relies exclusively on chemotherapy. Due to the lack of established molecular/genetic markers denoting parasite resistance, clinical treatment failure is often used as an indicator. Antimony-based drugs have been the standard antileishmanial treatment for more than seven decades, leading to major drug resistance in certain regions. Likewise, drug resistance to miltefosine and amphotericin B continues to spread at alarming rates. In consequence, innovative approaches are needed to accelerate the identification of antimicrobial drug targets and resistance mechanisms. To this end, we have implemented a novel approach based on thermal proteome profiling (TPP) to further characterize the mode of action of antileishmanials antimony, miltefosine and amphotericin B, as well as to better understand the mechanisms of drug resistance deployed by Leishmania. Proteins become more resistant to heat-induced denaturation when complexed with a ligand. In this way, we used multiplexed quantitative mass spectrometry-based proteomics to monitor the melting profile of thousands of expressed soluble proteins in WT, antimony-resistant, miltefosine-resistant, and amphotericin B-resistant L. infantum parasites, in the presence (or absence) of the above-mentioned drugs. Bioinformatics analyses were performed, including data normalization, melting profile fitting, and identification of proteins that underwent changes (fold change > 4) caused by complexation with a drug. With this unique approach, we were able to narrow down the regions of the L. infantum proteome that interact with antimony, miltefosine, and amphotericin B; validating previously-identified and unveiling novel drug targets. Moreover, analyses revealed candidate proteins potentially involved in drug resistance. Interestingly, we detected thermal proximity coaggregation for several proteins belonging to the same metabolic pathway (i.e., tryparedoxin peroxidase and aspartate aminotransferase in proteins exposed to antimony), highlighting the importance of these pathways. Collectively, our results could serve as a jumping-off point for the future development of innovative diagnostic tools for the detection and evaluation of antimicrobial-resistant Leishmania populations, as well as open the door for new on-target therapies.https://www.frontiersin.org/articles/10.3389/fcimb.2022.954144/fullLeishmaniaantileishmanial drugsmode of actionthermal proteome profilingdrug resistanceantimony |
| spellingShingle | Ana Victoria Ibarra-Meneses Ana Victoria Ibarra-Meneses Audrey Corbeil Audrey Corbeil Victoria Wagner Victoria Wagner Francis Beaudry Francis Beaudry Rubens L. do Monte-Neto Christopher Fernandez-Prada Christopher Fernandez-Prada Exploring direct and indirect targets of current antileishmanial drugs using a novel thermal proteomics profiling approach Frontiers in Cellular and Infection Microbiology Leishmania antileishmanial drugs mode of action thermal proteome profiling drug resistance antimony |
| title | Exploring direct and indirect targets of current antileishmanial drugs using a novel thermal proteomics profiling approach |
| title_full | Exploring direct and indirect targets of current antileishmanial drugs using a novel thermal proteomics profiling approach |
| title_fullStr | Exploring direct and indirect targets of current antileishmanial drugs using a novel thermal proteomics profiling approach |
| title_full_unstemmed | Exploring direct and indirect targets of current antileishmanial drugs using a novel thermal proteomics profiling approach |
| title_short | Exploring direct and indirect targets of current antileishmanial drugs using a novel thermal proteomics profiling approach |
| title_sort | exploring direct and indirect targets of current antileishmanial drugs using a novel thermal proteomics profiling approach |
| topic | Leishmania antileishmanial drugs mode of action thermal proteome profiling drug resistance antimony |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2022.954144/full |
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