Development and validation of a lung tri-culture spheroid model to evaluate therapeutic efficacy and toxicological safety against Cryptococcus neoformans.

• Grant number: 23/13538-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2024
• End date: December 31, 2024
• Field of knowledge: Health Sciences - Pharmacy
• Principal Investigator: Ana Marisa Fusco Almeida
• Grantee: Larissa Fernanda Bianchi
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

Cryptococcosis is a serious fungal infectious disease that is widespread and opportunistic, with significant public health implications, affecting both immunocompromised and immunocompetent individuals. The main causative agents are Cryptococcus neoformans and Cryptococcus gattii, which have ecological niches in pigeon droppings and decomposing tree matter, leading to their continuous dissemination. To gain a better understanding of the dynamics of C. neoformans interaction, this study aims to establish a 3D model of pulmonary spheroids using immortalized cell lines, characterized through morphological and microscopic techniques. The antifungal potential of nitrofurans will also be evaluated by treating infections in three-dimensional models and determining the model's viability. For this purpose, CALU-3, MRC-5, and THP-1 cells will be used for lung spheroid development. Cytotoxicity will be determined using the resazurin colorimetric method. In turn, the interaction between the spheroid and the fungus will be conducted with and without treatment using control drugs and nitrofurans, assessing fungal load. The data will undergo statistical analysis.