Role of cancer-associated fibroblasts on modulating regulated cell death of oral dysplastic keratinocytes: an in vitro study

• Grant number: 25/02099-1
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: May 01, 2025
• End date: April 30, 2026
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Luciana Corrêa
• Grantee: Saygo Tomo
• Supervisor: Daniela Elena Costea
• Host Institution: Faculdade de Odontologia (FO). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: University of Bergen, Norway
• Associated to the scholarship: 23/11402-4 - Regulated necrosis induced by photodynamic and sonodynamic therapies in oral epithelial dysplastic cells: in vitro and in vivo study, BP.PD

Abstract

The progression of oral epithelial dysplasia (OED) to oral squamous cell carcinoma (OSCC) involves intricate interactions between dysplastic keratinocytes and the tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs), particularly myofibroblastic CAFs (myCAFs) and inflammatory CAFs (iCAFs), may play a role in modulating regulated cell death (RCD) pathways, including pyroptosis, ferroptosis, and necroptosis. These pathways are critical for tumor control and may represent targets for oral cancer and oral epithelial dysplasia therapy but remain underexplored in the context of oral carcinogenesis. This study aims to investigate how CAFs influence RCD pathways in dysplastic oral keratinocytes (DOKs), providing insights into the molecular mechanisms driving OED progression. myCAFs will be induced by exposure of normal oral fibroblasts (NOFs) to TGF-¿ and iCAFs will be induced by co-culturing NOFs with OSCC cells (Luc-4) and validated via qPCR and Western blot. 3D organotypic models of OED incorporating myCAFs, iCAFs or NOFs will be exposed to inducers and inhibitors of pyroptosis, ferroptosis, and necroptosis. Histomorphometric and immunohistochemical analyses will be used to assess RCD endpoints, including markers such as cleaved N-terminal GSDMD for pyroptosis, 4-HNE for ferroptosis, and pMLKL for necroptosis.