title: Metabolic signatures of Besnoitia besnoiti-infected endothelial host cells and blockage of key metabolic pathways indicate high glycolytic and glutaminolytic needs of the parasite.
abstract: Besnoitia besnoiti is an obligate intracellular and emerging coccidian parasite of cattle with a significant economic impact on cattle industry. During acute infection, fast-proliferating tachyzoites are continuously formed mainly in endothelial host cells of infected animals. Given that offspring formation is a highly energy and cell building block demanding process, the parasite needs to exploit host cellular metabolism to meet its metabolic demands. Here, we analyzed the metabolic signatures of B. besnoiti-infected endothelial host cells and aimed to influence parasite proliferation by inhibitors of specific metabolic pathways. The following inhibitors were tested: fluoro 2-deoxy-D-glucose and 2-deoxy-D-glucose (FDG, DG; inhibitors of glycolysis), 6-diazo-5-oxo-L-norleucin (DON; inhibitor of glutaminolysis), dichloroacetate (DCA; inhibitor of pyruvate dehydrogenase kinase which favorites channeling of glucose carbons into the TCA cycle) and adenosine-monophosphate (AMP; inhibitor of ribose 5-P synthesis). Overall, B. besnoiti infections of bovine endothelial cells induced a significant and infection rate-dependent increase of glucose, lactate, glutamine, glutamate, pyruvate, alanine, and serine conversion rates which together indicate a parasite-triggered up-regulation of glycolysis and glutaminolysis. Thus, addition of DON, FDG, and DG into the cultivation medium of B. besnoiti infected endothelial cells led to a dose-dependent inhibition of parasite replication (4 μM DON, 99.5 % inhibition; 2 mM FDG, 99.1 % inhibition; 2 mM DG, 93 % inhibition; and 8 mM DCA, 71.9 % inhibition). In contrast, AMP had no significant effects on total tachyzoite production up to a concentration of 20 mM. Together, these data may open new strategies for the development of therapeutics for B. besnoiti infections.
authors: Institute of Parasitology, Biomedical Research Center Seltersberg, Justus Liebig University Giessen, Giessen, Germany. Anja.Taubert@vetmed.uni-giessen.de.
authors: A
authors: A
authors: Taubert
authors: Institute of Parasitology, Biomedical Research Center Seltersberg, Justus Liebig University Giessen, Giessen, Germany.
authors: C
authors: C
authors: Hermosilla
authors: Institute of Parasitology, Biomedical Research Center Seltersberg, Justus Liebig University Giessen, Giessen, Germany.
authors: L M R
authors: LM
authors: Silva
authors: Institute of Veterinary Physiology and Biochemistry, Justus Liebig University Giessen, Giessen, Germany.
authors: A
authors: A
authors: Wieck
authors: Unit for Biomathematics and Data Processing, Justus Liebig University Giessen, Giessen, Germany.
authors: K
authors: K
authors: Failing
authors: Institute of Veterinary Physiology and Biochemistry, Justus Liebig University Giessen, Giessen, Germany.
authors: S
authors: S
authors: Mazurek
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000378:metabolism; D002417:Cattle; D002418:Cattle Diseases / Q000378:metabolism / Q000469:parasitology*; D003048:Coccidiosis / Q000378:metabolism / Q000662:veterinary*; D004730:Endothelium, Vascular / Q000378:metabolism / Q000469:parasitology*; D005973:Glutamine / Q000378:metabolism*; D006019:Glycolysis; D066298:In Vitro Techniques; D053858:Metabolic Networks and Pathways / Q000187:drug effects; D021745:Sarcocystidae / Q000276:immunology / Q000378:metabolism*
chemical_list: D000913:Antibodies, Protozoan; D005973:Glutamine
keywords: 2-Deoxy-D-glucose; Coccidia; DON; Glutaminolysis; Glycolysis; Metabolic signature