Interaction of pathogenic microorganisms causing brain pathology, specifically Trypanosoma vivax, Zika virus, and SARS-CoV-2, with experimental hosts.

• Grant number: 25/01191-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2025
• End date: February 28, 2026
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Paola Marcella Camargo Minoprio
• Grantee: Guilherme Duarte Manfredi
• Host Institution: Institut Pasteur de São Paulo (IPSP). São Paulo , SP, Brazil
• Associated research grant: 17/27131-9 - Integrated strategy for the study of infectious agents causing emerging and/or neglected diseases transmitted by vectors of global impact, AP.TEM

Abstract

Most studies on cattle focus on milk and meat production; however, significant infectious diseases affecting the central nervous system, such as Nagana (bovine sleeping sickness) and bovine spongiform encephalopathy, have a considerable socio-economic impact. To date, in vitro models for studying bovine brain infections, particularly those involving the blood-brain barrier (BBB), are limited. The recent advancement in the development of bovine induced pluripotent stem cells (biPSCs) offers new possibilities for generating differentiated cells and tissues, including those that form the BBB. Our laboratory has successfully developed protocols for differentiating biPSCs into bovine neuroprogenitor cells (bNPCs), astrocytes, and endothelial cells. The identity of these cells was validated through the evaluation of gene expression markers specific to bNPCs (sox1, sox2, pax6, nestin, ki67, and musashi1), astrocytes (gfap, cd44, aquaporin4, s100-¿, and vimentin), and endothelial cells (brachyury, cdh5, gata-4, and pecam1). The goal of this project is to develop and optimize differentiation protocols for the construction of 2D and 3D in vitro models of the bovine BBB. For the 2D model, the transwell technique will be used, where bovine endothelial cells will be cultured in semi-permeable membrane inserts, allowing the characterization of their permeability, followed by the addition and co-culture of astrocytes and bNPCs to form the "cerebral" portion of the model. For the 3D model, bovine cerebral and vascular organoids will be developed, which will then be fused to generate vascularized bovine cerebral organoids. These 2D and 3D in vitro models will be applied to investigate the interactions between Trypanosoma vivax, the causative agent of Nagana, and bovine brain cells and tissues, as well as to analyze the effects of the parasite on BBB permeability. The development of these models will provide significant advancements in the understanding of bovine brain infection biology and will aid in the evaluation of potential chemotherapies.