title: Ability to grow on lipids accounts for the fully virulent phenotype in neutropenic mice of Aspergillus fumigatus null mutants in the key glyoxylate cycle enzymes.
abstract: Incidence and mortality rates of invasive aspergillosis clearly indicate the need of novel antifungals to treat patients suffering from this disease. Fungal proteins playing a crucial role in pathogenesis and with no orthologue in human cells are considered as primary therapeutic targets for the development of new antifungals with a high therapeutic index, one of the major drawbacks of the standard antifungal therapy, so far. In this work, we have analyzed the role in pathogenesis of the key enzymes of the Aspergillus fumigatus glyxoxylate cycle, isocitrate lyase and malate synthase, two possible candidates to primary therapeutic targets in this fungus. Deletion strains lacking isocitrate lyase (DeltaacuD strains) or malate synthase (DeltaacuE mutants) were constructed in this work. The Neurospora crassa pyr-4 gene was used as the replacing marker in gene deletion experiments. The pathogenicities of DeltaacuD and DeltaacuE mutants were tested in neutropenic mice and compared with those of two reference wild-type isolates A. fumigatus 237 and A. fumigatus 293. Interestingly, virulence and cytological studies clearly indicated the dispensability of the A. fumigatus glyoxylate cycle for pathogenicity. In addition, these results suggested the suitability of the pyr-4 gene as a valuable replacing marker for virulence studies in this fungus, a fact that was further confirmed by gene expression analyses. Finally, growth tests were performed to investigate the germination and growth of the DeltaacuD and DeltaacuE strains in nutrient deprivation environments, resembling the conditions that A. fumigatus conidia face after phagocytosis. Results obtained in this work strongly suggest that the ability to grow on lipids (triglycerides) of A. fumigatus isocitrate lyase and malate synthase deletion strains accounts for their fully virulent phenotype.
authors.affiliation: Departamento de Microbiología y Parasitología, Universidad de Alcalá, Carretera Madrid-Barcelona Km 33, Alcalá de Henares, ES-28871 Madrid, Spain.
authors.forename: Israel
authors.initials: I
authors.lastname: Olivas
authors.forename: Mar
authors.initials: M
authors.lastname: Royuela
authors.forename: Beatriz
authors.initials: B
authors.lastname: Romero
authors.forename: M Cândida
authors.initials: MC
authors.lastname: Monteiro
authors.forename: José M
authors.initials: JM
authors.lastname: Mínguez
authors.forename: Fernando
authors.initials: F
authors.lastname: Laborda
authors.forename: J Ramón
authors.initials: JR
authors.lastname: De Lucas
mesh_terms: D000818:Animals; D001228:Aspergillosis / Q000382:microbiology* / Q000473:pathology; D001232:Aspergillus fumigatus / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D003470:Culture Media; D006038:Glyoxylates / Q000378:metabolism; D007522:Isocitrate Lyase / Q000235:genetics / Q000378:metabolism; D008055:Lipids; D008292:Malate Synthase / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D009503:Neutropenia / Q000473:pathology; D017354:Point Mutation; D014774:Virulence
chemical_list: D003470:Culture Media; D006038:Glyoxylates; D008055:Lipids; D008292:Malate Synthase; D007522:Isocitrate Lyase; C031150:glyoxylic acid