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

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Winzer, Pablo and Anghel, Nicoleta and Imhof, Dennis and Balmer, Vreni and Ortega Mora, Luis Miguel and Ojo, Kayode K. and Van Voorhis, Wesley C. and Müller, Joachim and Hemphill, Andrew (2020) Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294. Pathogens, 9 (5). p. 382. ISSN 2076-0817

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Official URL: https://doi.org/10.3390/pathogens9050382




Abstract

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.


Item Type:Article
Uncontrolled Keywords:bumped kinase inhibitor; calcium-dependent protein kinase; immunofluorescence; immunogold labeling; inner membrane complex; neosporosis; surface antigen; tachyzoite; transmission electron microscopy
Subjects:Medical sciences > Veterinary > Microbiology
Medical sciences > Veterinary > Veterinary pharmacology
ID Code:65114
Deposited On:04 May 2021 15:41
Last Modified:05 May 2021 07:39

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