Analysis of Key Proteins of Cryptococcus neoformans Infection Differentially Expressed Biofilm/Planktonic in a Three-Dimensional Lung Model.

• Grant number: 24/15618-4
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: January 01, 2025
• End date: December 31, 2027
• Field of knowledge: Health Sciences - Pharmacy
• Principal Investigator: Ana Marisa Fusco Almeida
• Grantee: Marcos William de Lima Gualque
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

Cryptococcus neoformans is a yeast of medical relevance, especially in immunocompromised patients, such as those affected by AIDS. This fungus can cause serious and sometimes drug-resistant systemic infections, being responsible for more than 180,000 deaths worldwide every year. Infection occurs predominantly through inhalation of fungal propagules, leading to a pulmonary infection that can evolve systematically, mainly affecting the central nervous system. Virulence factors of this encapsulated yeast include the presence of a polysaccharide capsule, proteins such as melanin, phospholipase and urease, in addition to the ability to form biofilms and, consequently, highly resistant strains. Additionally, the formation of biofilms hinders phagocytosis and drug permeation, compromising the effectiveness of antifungal treatments. Three-dimensional cell culture models are promising for studying fungus-host interactions, providing an environment more representative of in vivo conditions and helping to develop new therapeutic interventions for cryptococcosis. With the purpose of evaluating proteins associated with the virulence of the Cryptococcus neoformans biofilm in a three-dimensional lung model, this study aims to evaluate the differential proteomic profile of the C. neoformans biofilm through infection in the three-dimensional lung model composed of the MRC-5, A549 lineages and THP-1, perform the quantification of transcripts of proteins differentially expressed in infected biofilm and planktonic in the three-dimensional model using mass spectrometry and bioinformatics tools, verify the relative expression of genes associated with proteins of interest by the qRT-PCR method, and finally validate these proteins in infection by comparing monolayer and three-dimensional models through the quantification of fluorescence with specific labels in multiphoton microscopy. With the data obtained, we intend to open a range of possibilities, from biofilm markers to possible therapeutic targets.