Influence of chondroitin sulfate on the activation of NLRP1 and NRLP3 inflammasomes in skin cells infected by a zoonotic agent cultured on collagen scaffold

• Grant number: 25/09140-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2025
• End date: June 30, 2026
• Field of knowledge: Agronomical Sciences - Veterinary Medicine
• Principal Investigator: Rodrigo Augusto da Silva
• Grantee: Cristina Ferri Calvi
• Host Institution: Vice-Reitoria de Pesquisa e Pós-Graduação. Universidade Paulista (UNIP). São Paulo , SP, Brazil

Abstract

The skin, being the largest organ in the body, acts as an interface between the body and the surrounding tissues. It plays important roles as a physical barrier in protecting against pathogens, using defensive strategies executed by resident immune cells to initiate inflammatory responses to a zoonotic fungal infection. In this scenario, we highlight the action of the NLRP3 inflammasome, recognized as a sensor of microbial signals and other danger signals, and of NRLP1, which has been linked to the activation of pyroptosis in epidermal cells. Thus, knowing that chondroitin sulfate can inhibit the production of excessive pro-inflammatory cytokines, such as IL-1¿ and TNF-¿, limiting tissue damage, this project aims to deepen knowledge about the involvement of NRLP1 and NRLP3 inflammasomes in zoonotic cutaneous fungal infections and their relationship with Chondroitin Sulfate. For this purpose, human keratinocyte (HaCaT) and human gingival fibroblast (HGF) cell lines will be cultured separately and cocultured on collagen substrate enriched or not with chondroitin sulfate and then exposed to the pathogens C. gattii and E. cuniculi (DOI 8:1) for 7 days. Then, biocompatibility and histopathological parameters will be evaluated by light and scanning electron microscopy. The cytokine concentration of the conditioned medium and the gene expression of the components of the NRLP1 and NRLP3 inflammasomes will also be evaluated by flow cytometry and qPCR, respectively. It is worth highlighting the need for better elucidation of the molecular mechanisms underlying epidermal fungal infection and we believe that the results obtained in this proposal can provide new and highly relevant information for this field of science that is still little explored.