Characterization of the role of chaperone-mediated autophagy in the cellular response to DNA damage by complementary methods

• Grant number: 24/08750-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2024
• End date: July 31, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Nicolas Carlos Hoch
• Grantee: Pedro Henrique de Almeida Carvalho
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Chaperone-mediated autophagy (CMA) is a selective process of intracellular protein degradation, in which a pentapeptide motif similar to KFERQ is recognized by cytosolic chaperones, that direct the substrate to the lysosomal surface, where the LAMP2A receptor allows cargo translocation across the lysosomal membrane. CMA plays a crucial role in maintaining cellular homeostasis, regulating multiple intracellular functions through specific protein depletion and contributing to the preservation of protein quality. CMA activity can be induced in response to stressors, such as nutrients deprivation, oxidative stress and DNA damage, with dysfunctions in this pathway being associated with diseases such as neurodegeneration and cancer. Preliminary results from our research group indicate that breast tumor cells knocked out for the LAMP2A isoform have greater resistance to agents that cause DNA damage. In this sense, this project aims to validate the phenotype observed in LAMP2A knockout cells, through complementation of these cells with wild-type LAMP2A using lentiviral vectors, in addition to confirming phenotypes through the use of LAMP2A-specific interference RNAs. Thus, this study will allow us to solidify and expand our initial findings in the search for a better understanding on the function of chaperone-mediated autophagy in the cellular response to DNA damage.