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Disruption of the inositol phosphorylceramide synthase gene affects Trypanosoma cruzi differentiation and infection capacity

Sphingolipids (SLs) are essential components of all eukaryotic cellular membranes. In fungi, plants and many protozoa, the primary SL is inositol-phosphorylceramide (IPC). Trypanosoma cruzi is a protozoan parasite that causes Chagas disease (CD), a chronic illness for which no vaccines or effective...

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Autores principales: Dos Santos, Nailma S A, Estevez-Castro, Carlos F., Macedo, Juan P., Chame, Daniela F., Castro-Gomes, Thiago, Santos-Cardoso, Mariana, Burle-Caldas, Gabriela A., Covington, Courtney N., Steel, Patrick G., Smith, Terry K., Denny, Paul W., Teixeira, Santuza M. R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10545103/
https://www.ncbi.nlm.nih.gov/pubmed/37729272
http://dx.doi.org/10.1371/journal.pntd.0011646
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author Dos Santos, Nailma S A
Estevez-Castro, Carlos F.
Macedo, Juan P.
Chame, Daniela F.
Castro-Gomes, Thiago
Santos-Cardoso, Mariana
Burle-Caldas, Gabriela A.
Covington, Courtney N.
Steel, Patrick G.
Smith, Terry K.
Denny, Paul W.
Teixeira, Santuza M. R.
author_facet Dos Santos, Nailma S A
Estevez-Castro, Carlos F.
Macedo, Juan P.
Chame, Daniela F.
Castro-Gomes, Thiago
Santos-Cardoso, Mariana
Burle-Caldas, Gabriela A.
Covington, Courtney N.
Steel, Patrick G.
Smith, Terry K.
Denny, Paul W.
Teixeira, Santuza M. R.
author_sort Dos Santos, Nailma S A
collection PubMed
description Sphingolipids (SLs) are essential components of all eukaryotic cellular membranes. In fungi, plants and many protozoa, the primary SL is inositol-phosphorylceramide (IPC). Trypanosoma cruzi is a protozoan parasite that causes Chagas disease (CD), a chronic illness for which no vaccines or effective treatments are available. IPC synthase (IPCS) has been considered an ideal target enzyme for drug development because phosphoinositol-containing SL is absent in mammalian cells and the enzyme activity has been described in all parasite forms of T. cruzi. Furthermore, IPCS is an integral membrane protein conserved amongst other kinetoplastids, including Leishmania major, for which specific inhibitors have been identified. Using a CRISPR-Cas9 protocol, we generated T. cruzi knockout (KO) mutants in which both alleles of the IPCS gene were disrupted. We demonstrated that the lack of IPCS activity does not affect epimastigote proliferation or its susceptibility to compounds that have been identified as inhibitors of the L. major IPCS. However, disruption of the T. cruzi IPCS gene negatively affected epimastigote differentiation into metacyclic trypomastigotes as well as proliferation of intracellular amastigotes and differentiation of amastigotes into tissue culture-derived trypomastigotes. In accordance with previous studies suggesting that IPC is a membrane component essential for parasite survival in the mammalian host, we showed that T. cruzi IPCS null mutants are unable to establish an infection in vivo, even in immune deficient mice.
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spelling pubmed-105451032023-10-03 Disruption of the inositol phosphorylceramide synthase gene affects Trypanosoma cruzi differentiation and infection capacity Dos Santos, Nailma S A Estevez-Castro, Carlos F. Macedo, Juan P. Chame, Daniela F. Castro-Gomes, Thiago Santos-Cardoso, Mariana Burle-Caldas, Gabriela A. Covington, Courtney N. Steel, Patrick G. Smith, Terry K. Denny, Paul W. Teixeira, Santuza M. R. PLoS Negl Trop Dis Research Article Sphingolipids (SLs) are essential components of all eukaryotic cellular membranes. In fungi, plants and many protozoa, the primary SL is inositol-phosphorylceramide (IPC). Trypanosoma cruzi is a protozoan parasite that causes Chagas disease (CD), a chronic illness for which no vaccines or effective treatments are available. IPC synthase (IPCS) has been considered an ideal target enzyme for drug development because phosphoinositol-containing SL is absent in mammalian cells and the enzyme activity has been described in all parasite forms of T. cruzi. Furthermore, IPCS is an integral membrane protein conserved amongst other kinetoplastids, including Leishmania major, for which specific inhibitors have been identified. Using a CRISPR-Cas9 protocol, we generated T. cruzi knockout (KO) mutants in which both alleles of the IPCS gene were disrupted. We demonstrated that the lack of IPCS activity does not affect epimastigote proliferation or its susceptibility to compounds that have been identified as inhibitors of the L. major IPCS. However, disruption of the T. cruzi IPCS gene negatively affected epimastigote differentiation into metacyclic trypomastigotes as well as proliferation of intracellular amastigotes and differentiation of amastigotes into tissue culture-derived trypomastigotes. In accordance with previous studies suggesting that IPC is a membrane component essential for parasite survival in the mammalian host, we showed that T. cruzi IPCS null mutants are unable to establish an infection in vivo, even in immune deficient mice. Public Library of Science 2023-09-20 /pmc/articles/PMC10545103/ /pubmed/37729272 http://dx.doi.org/10.1371/journal.pntd.0011646 Text en © 2023 Dos Santos et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Dos Santos, Nailma S A
Estevez-Castro, Carlos F.
Macedo, Juan P.
Chame, Daniela F.
Castro-Gomes, Thiago
Santos-Cardoso, Mariana
Burle-Caldas, Gabriela A.
Covington, Courtney N.
Steel, Patrick G.
Smith, Terry K.
Denny, Paul W.
Teixeira, Santuza M. R.
Disruption of the inositol phosphorylceramide synthase gene affects Trypanosoma cruzi differentiation and infection capacity
title Disruption of the inositol phosphorylceramide synthase gene affects Trypanosoma cruzi differentiation and infection capacity
title_full Disruption of the inositol phosphorylceramide synthase gene affects Trypanosoma cruzi differentiation and infection capacity
title_fullStr Disruption of the inositol phosphorylceramide synthase gene affects Trypanosoma cruzi differentiation and infection capacity
title_full_unstemmed Disruption of the inositol phosphorylceramide synthase gene affects Trypanosoma cruzi differentiation and infection capacity
title_short Disruption of the inositol phosphorylceramide synthase gene affects Trypanosoma cruzi differentiation and infection capacity
title_sort disruption of the inositol phosphorylceramide synthase gene affects trypanosoma cruzi differentiation and infection capacity
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10545103/
https://www.ncbi.nlm.nih.gov/pubmed/37729272
http://dx.doi.org/10.1371/journal.pntd.0011646
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