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1.
Methods Mol Biol ; 2775: 47-55, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38758310

RESUMO

In vivo models provide advantages to study the progression of disease and to identify potential biomarkers to detect and monitor infections. For the human fungal pathogen Cryptococcus neoformans, murine intranasal models aim to recapitulate natural infection from inhalation of desiccated fungal cells from the environment and permit monitoring of disease over time. In this chapter, we describe the establishment of a murine model for cryptococcosis and the subsequent collection of organs, tissues, and fluids for sampling. These samples may support novel diagnostic strategies and opportunities to monitor dissemination of the fungal cells throughout the host and propose new treatment options to combat disease.


Assuntos
Criptococose , Cryptococcus neoformans , Modelos Animais de Doenças , Animais , Cryptococcus neoformans/patogenicidade , Criptococose/microbiologia , Criptococose/diagnóstico , Camundongos , Manejo de Espécimes/métodos , Humanos
2.
Methods Mol Biol ; 2775: 127-137, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38758315

RESUMO

Proteomic profiling provides in-depth information about the regulation of diverse biological processes, activation of and communication across signaling networks, and alterations to protein production, modifications, and interactions. For infectious disease research, mass spectrometry-based proteomics enables detection of host defenses against infection and mechanisms used by the pathogen to evade such responses. In this chapter, we outline protein extraction from organs, tissues, and fluids collected following intranasal inoculation of a murine model with the human fungal pathogen Cryptococcus neoformans. We describe sample preparation, followed by purification, processing on the mass spectrometer, and a robust bioinformatics analysis. The information gleaned from proteomic profiling of fungal infections supports the detection of novel biomarkers for diagnostic and prognostic purposes.


Assuntos
Criptococose , Cryptococcus neoformans , Modelos Animais de Doenças , Proteômica , Animais , Cryptococcus neoformans/metabolismo , Cryptococcus neoformans/patogenicidade , Camundongos , Criptococose/microbiologia , Criptococose/metabolismo , Proteômica/métodos , Biologia Computacional/métodos , Proteoma/metabolismo , Biomarcadores/metabolismo , Espectrometria de Massas/métodos
3.
Int J Mol Sci ; 22(22)2021 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-34830272

RESUMO

Fungal pathogens cause an array of diseases by targeting both immunocompromised and immunocompetent hosts. Fungi overcome our current arsenal of antifungals through the emergence and evolution of resistance. In particular, the human fungal pathogen, Cryptococcus neoformans is found ubiquitously within the environment and causes severe disease in immunocompromised individuals around the globe with limited treatment options available. To uncover fundamental knowledge about this fungal pathogen, as well as investigate new detection and treatment strategies, mass spectrometry-based proteomics provides a plethora of tools and applications, as well as bioinformatics platforms. In this review, we highlight proteomics approaches within the laboratory to investigate changes in the cellular proteome, secretome, and extracellular vesicles. We also explore regulation by post-translational modifications and the impact of protein-protein interactions. Further, we present the development and comprehensive assessment of murine models of cryptococcal infection, which provide valuable tools to define the dynamic relationship between the host and pathogen during disease. Finally, we explore recent quantitative proteomics studies that begin to extrapolate the findings from the bench to the clinic for improved methods of fungal detection and monitoring. Such studies support a framework for personalized medical approaches to eradicate diseases caused by C. neoformans.


Assuntos
Criptococose/metabolismo , Cryptococcus neoformans/metabolismo , Proteínas Fúngicas/metabolismo , Proteoma/metabolismo , Proteômica/métodos , Animais , Antifúngicos/uso terapêutico , Criptococose/tratamento farmacológico , Criptococose/microbiologia , Cryptococcus neoformans/genética , Cryptococcus neoformans/patogenicidade , Modelos Animais de Doenças , Farmacorresistência Fúngica/genética , Vesículas Extracelulares/metabolismo , Proteínas Fúngicas/genética , Interações Hospedeiro-Parasita/genética , Humanos , Camundongos , Medicina de Precisão/métodos , Mapas de Interação de Proteínas/genética , Processamento de Proteína Pós-Traducional/genética , Proteoma/genética , Secretoma/metabolismo , Transcriptoma , Resultado do Tratamento , Fatores de Virulência/metabolismo
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