Cyclic Antifungal Treatments as Modulators of Extracellular Vesicle Biogenesis and Protein Corona Formation in Biological Fluids

• Grant number: 25/07930-0
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: September 01, 2025
• End date: August 31, 2026
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Fausto Bruno dos Reis Almeida
• Grantee: Lucas Fabrício Bahia Nogueira
• Supervisor: Van Dijck Patrick
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Institution abroad: University of Leuven, Leuven (KU Leuven), Belgium
• Associated to the scholarship: 24/00403-2 - Deciphering the Functional Role of Protein Corona in Cellular Communication mediated by Fungal Extracellular Vesicles, BP.PD

Abstract

Fungal infections caused by Candida auris (C. auris) and Aspergillus fumigatus (A. fumigatus) represent a critical global health challenge due to their high mortality rates and increasing antifungal resistance. Extracellular vesicles (EVs) play a central role in fungal adaptation, facilitating biofilm formation, immune evasion, and drug resistance. Upon exposure to biological fluids, EVs acquire a protein corona-a dynamic layer of host-derived proteins-that modulates their stability, biodistribution, and immune recognition. This corona consists of primary and secondary layers, both influencing fungal adaptation and host-pathogen interactions. However, the impact of antifungal treatments on EV biogenesis and protein corona composition remains poorly understood. In this context, we hypothesize that cyclic treatment with amphotericin B (AMB) and caspofungin (CAS) alters EV secretion, molecular cargo, and protein corona dynamics, modulating EV interactions with epithelial and immune cells. This novel hypothesis explores how antifungal treatments not only impact EV secretion but also alter the molecular cargo and structure of the protein corona. By investigating the primary and secondary coronas, this study aims to clarify how antifungal agents influence fungal EV biogenesis, with potential implications for developing therapeutic strategies targeting these vesicles. The primary objectives are to: (i) isolate and characterize EVs from C. auris and A. fumigatus under different culture conditions, analyzing their protein corona profiles; (ii) identify proteins in the primary and secondary coronas using high-resolution proteomic analysis; (iii) assess how AMB/CAS treatment influences EV biogenesis and protein corona composition; and (iv) Evaluate the functional impact of EVs-with and without the protein corona-on epithelial and immune cell interactions using in vivo models, such as Galleria mellonella and murine intravenous infection models, and potentially through organ-on-a-chip platforms in future collaborative efforts, if available. By integrating high-resolution analytical techniques with validation using both in vivo models, this study bridges the gap between ex vivo and in vivo protein corona dynamics. The findings will provide novel insights into fungal EV-mediated resistance mechanisms and immune modulation, contributing to the development of EV-targeted therapies to combat antifungal resistance and improve immune responses to fungal infections.