Analysis of the role of p53 and NRF2 transcription factors in the modulation of cell death by ferroptosis in glioblastoma cell lines

• Grant number: 25/10646-2
• Support Opportunities: Scholarships in Brazil - Master
• Start date: October 01, 2025
• End date: September 30, 2027
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Clarissa Ribeiro Reily Rocha
• Grantee: Juliana Valério de Oliveira
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

Analysis of the Role of the Transcription Factors p53 and NRF2 in Regulating Ferroptotic Cell Death in Glioblastoma Cell LinesGlioblastoma is the most aggressive type of brain tumor and is responsible for the majority of deaths among central nervous system cancers. It has an almost 100% recurrence rate. Although there are treatment options like surgery, drug therapy, and chemotherapy with temozolomide (TMZ), they show limited success in improving patient survival and rarely prevent the tumor from returning. Even though TMZ is the main treatment used, over time it leads to resistance to the medication.One way to try to reduce this resistance is by using compounds that trigger non-apoptotic types of cell death, such as ferroptosis. Ferroptosis is a form of programmed cell death that depends on iron and is marked by damage to lipids in the cell membrane. It has gained attention as a promising strategy to overcome drug resistance in tumors. NRF2 is a transcription factor that helps cells respond to oxidative stress and can block ferroptosis by activating the xCT transporter. On the other hand, p53 can make cells more sensitive to ferroptosis by suppressing this same transporter.This study aims to understand how NRF2 and p53 interact in controlling xCT and how that affects glioblastoma cells' sensitivity to ferroptosis. To do this, we will use glioblastoma cell lines with either normal p53 (U87MG, U343MG) or mutated p53 (U251MG, U138MG), as well as genetically modified lines with increased or deleted expression of p53 and NRF2. We will evaluate how sensitive these cells are to ferroptosis using cell viability tests (XTT), colony formation assays, and measurements of lipid damage (with BODIPY 581/591 C11), after treatment with ferroptosis inducers (erastin, RSL3, ML210) and an NRF2 activator (Ki696). We will also measure the gene and protein levels of NRF2, SLC7A11, and p53 using RT-qPCR and Western blot, along with levels of glutathione (GSH) and reactive oxygen species (ROS).Understanding how NRF2 and p53 work together could reveal important mechanisms behind glioblastoma resistance. By gaining more insight into this interaction, the study hopes to contribute to the development of new treatments that target these factors to fight tumor growth and resistance to conventional treatment methods.