Exploring the Complexities of Neurocryptococcosis: Investigating the Dynamics between Blood-Brain Barrier, Pharmacokinetics, and Inflammation.

• Grant number: 24/03190-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: November 01, 2024
• End date: October 31, 2026
• Field of knowledge: Biological Sciences - Pharmacology - Clinical Pharmacology
• Principal Investigator: João Paulo Bianchi Ximenez
• Grantee: Cecília Cristina de Souza Rocha
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Neurocryptococcosis is a subacute meningoencephalitis caused by the fungus Cryptococcus spp., occurring most commonly in patients diagnosed with HIV or other forms of immunosuppression. Amphotericin B (AnfB), the drug of choice for the treatment of neurocryptococcosis, has low permeability in the blood-brain barrier (BBB), with consequent low concentrations in the cerebrospinal fluid (CSF). P-glycoprotein (P-gp) is widely expressed in the BBB and functions as an important pathway for the efflux of AnfB into the systemic circulation; consequently, its inhibition increases the permeability of AnfB in the CNS. The integrity of the BBB and the composition of the CSF may change in situations resulting from infectious and/or inflammatory diseases. Considering that inflammation alters the expression of membrane transporters, and consequently the pharmacokinetic parameters, the inflammatory response can be considered an important factor in the interindividual variability of the efficacy and toxicity of drugs. The main objective of the present study is to evaluate the influence of pro-inflammatory cytokines and extracellular vesicles from Cryptococcus neoformans on the integrity of the BBB and on AnfB concentrations using the hCMEC/D3 in vitro model. Therefore, (1) we will develop methods for analyzing AnfB in the hCMEC/D3 cell line lysate by LC-MS/MS; (2) we will evaluate the impact of pro-inflammatory cytokines (TNF-±, IL-1², IL-6) on the pharmacokinetics of AnfB in the hCMEC/D3 cell line; (3) we will establish the influence of C. neoformans extracellular vesicles on the pharmacokinetics of AnfB in the hCMEC/D3 cell line; (4) we will determine changes in the pharmacokinetics of AnfB in the presence of pro-inflammatory cytokines (TNF-±, IL-1², IL-6) and C. neoformans extracellular vesicles in the hCMEC/D3 cell line. With the results, we can understand how the inflammatory and infectious process can act on the integrity of the BBB and the pharmacokinetics of AnfB in both plasma and CSF of patients with neurocryptococcosis.