<i>Drosophila melanogaster</i> as a model to study virulence and azole treatment of the emerging pathogen <i>Candida auris</i>

Wurster, Sebastian; Bandi, Ashwini; Beyda, Nicholas D.; Albert, Nathaniel D.; Raman, Nitya Meenakshi; Raad, Isaam; Kontoyiannis, Dimitrios P.

doi:10.1093/jac/dkz100

Cited by 40 publications

(37 citation statements)

References 23 publications

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“…This new model system allows for fungal burden to be monitored throughout the course of infection in a single animal via CFU count. This is unlike any other experimental system, where fungal burden is either an endpoint measure in the mouse kidney, in homogenized wax moth larvae [71], nematodes [72], flies [73], or requires genetically modified fluorescent yeast strains for microscopic imaging and analyses [74]. While we did detect very low CFU counts in the PBS control 3 days post infection, we consider this a spurious finding due to cross contamination as preliminary experiments of plating contents of hemolymph and feces of naïve animals did not detect any yeast growth (data not shown).…”

Section: Discussionmentioning

confidence: 99%

Tobacco Hornworm (Manduca sexta) caterpillars as a novel host model for the study of fungal virulence and drug efficacy

et al. 2020

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The two leading yeast pathogens of humans, Candida albicans and Cryptococcus neoformans, cause systemic infections in >1.4 million patients worldwide with mortality rates approaching 75%. It is thus imperative to study fungal virulence mechanisms, efficacy of antifungal drugs, and host response pathways. While this is commonly done in mammalian models, which are afflicted by ethical and practical concerns, invertebrate models, such as wax moth larvae and nematodes have been introduced over the last two decades. To complement existing invertebrate host models, we developed fifth instar caterpillars of the Tobacco Hornworm moth Manduca sexta as a novel host model. These caterpillars can be maintained at 37°C, are suitable for injections with defined amounts of yeast cells, and are susceptible to the most threatening yeast pathogens, including C. albicans, C. neoformans, C. auris, and C. glabrata. Importantly, fungal burden can be assessed daily throughout the course of infection in a single caterpillar's feces and hemolymph. Infected caterpillars can be rescued by treatment with antifungal drugs. Notably, these animals are large enough for weight to provide a reliable and reproducible measure of fungal disease and to facilitate host tissue-specific expression analyses. M. sexta caterpillars combine a suite of parameters that make them suitable for the study of fungal virulence.

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Section: Discussionmentioning

confidence: 99%

Tobacco Hornworm (Manduca sexta) caterpillars as a novel host model for the study of fungal virulence and drug efficacy

et al. 2020

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“…albicans virulence include a Drosophila infection model (Alarco et al 2004;Chamilos et al 2006;Glittenberg et al 2011;Wurster et al 2019), Galleria mellonella moth larvae model (Fuchs et al 2010;Frenkel et al 2016). Very recently, Manduca sexta caterpillars have been developed as a novel host model for the study of fungal virulence and drug efficacy (Lyons et al 2020), which have several advantages over other non-mammalian models, including their ability to be maintained at 37˚C and ability to assess fungal burden throughout the course of infection via the caterpillar's hemolymph of feces.…”

Section: Discussionmentioning

confidence: 99%

Systematic characterization and genetic interaction analysis of adhesins inCandida albicansvirulence

Rosiana

Zhang

Kim

et al. 2020

Preprint

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Candida albicans is a microbial fungus that exists as a commensal member of the human microbiome and an opportunistic pathogen. Cell surface-associated adhesin proteins play a crucial role in C. albicans' ability to undergo cellular morphogenesis, develop robust biofilms, colonize, and cause infection in a host. However, a comprehensive analysis of the role and relationships between these adhesins has not been explored. We previously established a CRISPR-based platform for efficient generation of single- and double-gene deletions in C. albicans, which was used to construct a library of 144 mutants, comprising 12 unique adhesin genes deleted singly, or in every possible combination of double deletions. Here, we exploit this adhesin mutant library to explore the role of adhesin proteins in C. albicans virulence. We perform a comprehensive, high-throughput screen of this library, using Caenorhabditis elegans as a simplified model host system, which identified mutants critical for virulence and significant genetic interactions. We perform follow-up analysis to assess the ability of high- and low-virulence strains to undergo cellular morphogenesis and form biofilms in vitro, as well as to colonize the C. elegans host. We further perform genetic interaction analysis to identify novel significant negative genetic interactions between adhesin mutants, whereby combinatorial perturbation of these genes significantly impairs virulence, more than expected based on virulence of the single mutant constituent strains. Together, this yields important new insight into the role of adhesins, singly and in combinations, in mediating diverse facets of virulence of this critical fungal pathogen.

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“…This new model system allows for fungal burden to be monitored throughout the course of infection in a single animal via CFU count. This is unlike any other experimental system, where fungal burden is either an endpoint measure in the mouse kidney, in homogenised was moth larvae 49 ,nematodes 50 , flies 51 , or requires genetically modified fluorescent yeast strains for microscopic imaging and analyses 52 . While we did detect very low CFU counts in the PBS control 3 days post infection, we consider this spurious finding due to cross contamination as preliminary experiments of plating contents of hemolymph and faeces of naïve animals did not detect any yeast growth (data not shown).…”

Section: Discussionmentioning

confidence: 99%

Tobacco Hornworm (Manduca sexta) caterpillars as a novel host model for the study of fungal virulence and drug efficacy

Lyons

Softley

Balfour

et al. 2019

Preprint

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29Pathogenic yeast species can cause life-threatening infections in humans. The two leading 30 yeast pathogens, Candida albicans and Cryptococcus neoformans, cause systemic infections 31 in >1.4 million patients world-wide with mortality rates approaching 75%. It is thus 32 imperative to study fungal virulence mechanisms, stress response pathways, and the efficacy 33 of antifungal drugs. This is commonly done using mammalian models. To address ethical and 34 practical concerns, invertebrate models, such as wax moth larvae, nematodes, or flies, have 35 been introduced over the last two decades. To address short-comings in existing invertebrate 36 host models, we developed fifth instar caterpillars of the Tobacco Hornworm moth Manduca 37 sexta as a novel host model for the study of fungal virulence and drug efficacy. These 38 caterpillars can be raised at standardised conditions, maintained at 37˚C, can be injected with 39 defined amounts of yeast cells, and are susceptible to the most threatening yeast pathogens, 40 including C. albicans, C. neoformans, C. auris, and C. glabrata. Infected caterpillars can be 41 rescued by treatment with commonly deployed antifungal drugs and importantly, fungal 42 burden can be assessed daily throughout the course of infection in a single caterpillar's faeces 43 and hemolymph. Notably, these animals are large enough so that weight provides a reliable 44 and reproducible measure of fungal virulence. This model combines a suite of parameters that 45 recommend it for the study of fungal virulence. 46 47 50 tuberculosis 1 . The leading yeast pathogens, Candida albicans and Cryptococcus neoformans, 51 together account for >1,400,000 life-threatening infections world-wide with mortality rates 52 approaching 75% 1 . Candidemia, most commonly caused by C. albicans, is the fourth most 53 common cause of nosocomial blood stream infections, only surpassed by infections with 54Staphylococci and Enterococcus spp. Disturbingly, candidemia incidence rates are on the 55 rise. Within less than ten years, incidence rates increased by 36% 2 . Although cryptococcosis 56 incidence rates are on the decline in North America, this AIDS-defining illness is responsible 57 for 15% of all AIDS-related deaths world-wide 2,3 . This already dire situation is further 58 confounded by the emergence of drug resistant yeast species. Patients at risk of developing 59 invasive candidemia are often prophylactically treated with fluconazole, while the 60 echinocandins are considered a first line defence strategy 4 . Yet, C. glabrata, the most 61 3 common non-albicans Candida species associated with nosocomial blood stream infections 5 , 62 is intrinsically less susceptible to azole drugs and acquires resistance to echinocandins 63 rapidly 6 . The rapid global spread of multi-drug resistant C. auris has further exacerbated the 64 threat posed by fungi. C. auris was first reported in 2009 in Japan 7 . In 2015, C. auris arrived 65 in Europe causing an outbreak involving 72 patients in a cardio-thoracic hospital i...

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Drosophila melanogaster as a model to study virulence and azole treatment of the emerging pathogen Candida auris

Cited by 40 publications

References 23 publications

Tobacco Hornworm (Manduca sexta) caterpillars as a novel host model for the study of fungal virulence and drug efficacy

Tobacco Hornworm (Manduca sexta) caterpillars as a novel host model for the study of fungal virulence and drug efficacy

Systematic characterization and genetic interaction analysis of adhesins inCandida albicansvirulence

Tobacco Hornworm (Manduca sexta) caterpillars as a novel host model for the study of fungal virulence and drug efficacy

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