Study of the Regulation of RECK Expression by miRNAs in Cells Expressing HPV Genes

• Grant number: 23/12750-6
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: March 01, 2024
• End date: February 29, 2028
• Field of knowledge: Biological Sciences - Microbiology
• Principal Investigator: Enrique Mario Boccardo Pierulivo
• Grantee: Amanda Campos Nocera
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Persistent infection with Human Papillomavirus is etiologically associated with a set of pathologies, especially cervical cancer. One of the scenarios that allows the establishment and maintenance of these diseases is alterations in the tumor microenvironment. Research from our group indicates that the expression of RECK, a membrane-localized protein that regulates the activity of metalloproteinases in the microenvironment, is decreased in clinical samples of cervical cancers and their derived cell lines. This scenario favors disease progression. However, the mechanisms that lead to the negative modulation of the levels of this protein are not yet fully understood. In the present study, the involvement of miRNAs in the negative regulation of the gene encoding this protein will be investigated, which were selected through bioinformatics analyses previously conducted by our research group.Firstly, the expression profile of these molecules will be evaluated in cell lines derived from these tumors, as well as the expression of RECK through gain or loss of function of the miRNAs. Next, the negative regulation of the gene by miRNAs will be validated through a gene reporter assay using vectors containing the mutated 3'UTR region of RECK for the selected miRNA binding sites downstream of the luciferase gene. Finally, the effects on cell death, proliferation, invasion, and migration will be assessed when there is a gain or loss of miRNA function. Additionally, we will analyze the role of the chosen miRNAs in communication with other cells in the microenvironment, such as fibroblasts and endothelial cells, through exosomes.