Therapeutic potential of the Nrf2/ARE signaling pathway in doxorubicin-induced cardiotoxicity

Línea de investigación

Mitochondrial Pathophysiology.

Descripción

The anthracycline anticancer drug doxorubicin (Dox) inhibits DNA replication and cellular metabolic processes in cancer cells with high proliferative potential, causing myocardial damage and heart failure as side effects, which limits its use as a chemotherapeutic agent. The antioxidant transcription factor Nrf2 (nuclear factor erythroid 2-related factor 2) is a redox-sensitive factor that plays an essential role in the cellular defense against oxidative stress. Nrf2 mediates the cellular response to oxidants and electrophiles by binding to an antioxidant response element (ARE) in the promoter region of cytoprotective genes. Under basal conditions, Nrf2 is sequestered in the cytoplasm by Keap1 (Kelch-like ECH-associated protein 1) that ubiquitinates and targets Nrf2 for continuous proteasomal degradation. The activation of Nrf2 could protect against Dox-induced cardiotoxicity not only by decreasing reactive oxygen species (ROS) generation but also by affecting cellular metabolism, as well as mitochondrial structure and function. To test this hypothesis, we will perform a Dox dose-response experiment for ROS generation and cell death using fluorescent probes (CellROX Deep Red and DAPI) by flow cytometry and fluorescence microscopy. We will use activators of Nrf2 (dimethyl fumarate, sulforaphane, carnosic acid), and the novel protein-protein interaction (PPI) inhibitors ML 334 and RA 839 that disrupt the Nrf2-Keap1 interaction. We will also determine mitochondrial function using the Agilent Seahorse technology. Additionally, we will constitutively activate Nrf2 in cardiomyocytes by suppressing Keap1 expression. Mitochondrial ultrastructure will be analyzed by transmission electron microscopy. The results of this study will help understand the mechanisms by which Dox causes toxicity to the heart, and may help to develop new therapeutic strategies based on Nrf2 activation.

Contacto

Susana Cadenas Álvarez.

Correo electrónico: scadenas@cbm.csic.es.

Centro de Biología Molecular Severo Ochoa (CBM).

Número de plazas ofertadas: 1.