Cargando…

Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294

Background: Bumped kinase inhibitors (BKIs) are potential drugs for neosporosis treatment in farm animals. BKI-1294 exposure results in the formation of multinucleated complexes (MNCs), which remain viable in vitro under constant drug pressure. We investigated the formation of BKI-1294 induced MNCs,...

Descripción completa

Detalles Bibliográficos
Autores principales: Winzer, Pablo, Anghel, Nicoleta, Imhof, Dennis, Balmer, Vreni, Ortega-Mora, Luis-Miguel, Ojo, Kayode K., Van Voorhis, Wesley C., Müller, Joachim, Hemphill, Andrew
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7281336/
https://www.ncbi.nlm.nih.gov/pubmed/32429314
http://dx.doi.org/10.3390/pathogens9050382
_version_ 1783543899641348096
author Winzer, Pablo
Anghel, Nicoleta
Imhof, Dennis
Balmer, Vreni
Ortega-Mora, Luis-Miguel
Ojo, Kayode K.
Van Voorhis, Wesley C.
Müller, Joachim
Hemphill, Andrew
author_facet Winzer, Pablo
Anghel, Nicoleta
Imhof, Dennis
Balmer, Vreni
Ortega-Mora, Luis-Miguel
Ojo, Kayode K.
Van Voorhis, Wesley C.
Müller, Joachim
Hemphill, Andrew
author_sort Winzer, Pablo
collection PubMed
description Background: Bumped kinase inhibitors (BKIs) are potential drugs for neosporosis treatment in farm animals. BKI-1294 exposure results in the formation of multinucleated complexes (MNCs), which remain viable in vitro under constant drug pressure. We investigated the formation of BKI-1294 induced MNCs, the re-emergence of viable tachyzoites following drug removal, and the localization of CDPK1, the molecular target of BKIs. Methods: N. caninum tachyzoites and MNCs were studied by TEM and immunofluorescence using antibodies directed against CDPK1, and against NcSAG1 and IMC1 as markers for tachyzoites and newly formed zoites, respectively. Results: After six days of drug exposure, MNCs lacked SAG1 surface expression but remained intracellular, and formed numerous zoites incapable of disjoining from each other. Following drug removal, proliferation continued, and zoites lacking NcSAG1 emerged from the periphery of these complexes, forming infective tachyzoites after 10 days. In intracellular tachyzoites, CDPK1 was evenly distributed but shifted towards the apical part once parasites were extracellular. This shift was not affected by BKI-1294. Conclusions: CDPK1 has a dynamic distribution depending on whether parasites are located within a host cell or outside. During MNC-to-tachyzoite reconversion newly formed tachyzoites are generated directly from MNCs through zoites of unknown surface antigen composition. Further in vivo studies are needed to determine if MNCs could lead to a persistent reservoir of infection after BKI treatment.
format Online
Article
Text
id pubmed-7281336
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-72813362020-06-19 Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294 Winzer, Pablo Anghel, Nicoleta Imhof, Dennis Balmer, Vreni Ortega-Mora, Luis-Miguel Ojo, Kayode K. Van Voorhis, Wesley C. Müller, Joachim Hemphill, Andrew Pathogens Article Background: Bumped kinase inhibitors (BKIs) are potential drugs for neosporosis treatment in farm animals. BKI-1294 exposure results in the formation of multinucleated complexes (MNCs), which remain viable in vitro under constant drug pressure. We investigated the formation of BKI-1294 induced MNCs, the re-emergence of viable tachyzoites following drug removal, and the localization of CDPK1, the molecular target of BKIs. Methods: N. caninum tachyzoites and MNCs were studied by TEM and immunofluorescence using antibodies directed against CDPK1, and against NcSAG1 and IMC1 as markers for tachyzoites and newly formed zoites, respectively. Results: After six days of drug exposure, MNCs lacked SAG1 surface expression but remained intracellular, and formed numerous zoites incapable of disjoining from each other. Following drug removal, proliferation continued, and zoites lacking NcSAG1 emerged from the periphery of these complexes, forming infective tachyzoites after 10 days. In intracellular tachyzoites, CDPK1 was evenly distributed but shifted towards the apical part once parasites were extracellular. This shift was not affected by BKI-1294. Conclusions: CDPK1 has a dynamic distribution depending on whether parasites are located within a host cell or outside. During MNC-to-tachyzoite reconversion newly formed tachyzoites are generated directly from MNCs through zoites of unknown surface antigen composition. Further in vivo studies are needed to determine if MNCs could lead to a persistent reservoir of infection after BKI treatment. MDPI 2020-05-16 /pmc/articles/PMC7281336/ /pubmed/32429314 http://dx.doi.org/10.3390/pathogens9050382 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Winzer, Pablo
Anghel, Nicoleta
Imhof, Dennis
Balmer, Vreni
Ortega-Mora, Luis-Miguel
Ojo, Kayode K.
Van Voorhis, Wesley C.
Müller, Joachim
Hemphill, Andrew
Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title_full Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title_fullStr Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title_full_unstemmed Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title_short Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title_sort neospora caninum: structure and fate of multinucleated complexes induced by the bumped kinase inhibitor bki-1294
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7281336/
https://www.ncbi.nlm.nih.gov/pubmed/32429314
http://dx.doi.org/10.3390/pathogens9050382
work_keys_str_mv AT winzerpablo neosporacaninumstructureandfateofmultinucleatedcomplexesinducedbythebumpedkinaseinhibitorbki1294
AT anghelnicoleta neosporacaninumstructureandfateofmultinucleatedcomplexesinducedbythebumpedkinaseinhibitorbki1294
AT imhofdennis neosporacaninumstructureandfateofmultinucleatedcomplexesinducedbythebumpedkinaseinhibitorbki1294
AT balmervreni neosporacaninumstructureandfateofmultinucleatedcomplexesinducedbythebumpedkinaseinhibitorbki1294
AT ortegamoraluismiguel neosporacaninumstructureandfateofmultinucleatedcomplexesinducedbythebumpedkinaseinhibitorbki1294
AT ojokayodek neosporacaninumstructureandfateofmultinucleatedcomplexesinducedbythebumpedkinaseinhibitorbki1294
AT vanvoorhiswesleyc neosporacaninumstructureandfateofmultinucleatedcomplexesinducedbythebumpedkinaseinhibitorbki1294
AT mullerjoachim neosporacaninumstructureandfateofmultinucleatedcomplexesinducedbythebumpedkinaseinhibitorbki1294
AT hemphillandrew neosporacaninumstructureandfateofmultinucleatedcomplexesinducedbythebumpedkinaseinhibitorbki1294