Development of RNA-based therapeutic approaches

• Grant number: 23/02980-4
• Support Opportunities: Regular Research Grants
• Start date: October 01, 2023
• End date: September 30, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Patricia Pereira Coltri
• Grantee: Patricia Pereira Coltri
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Arthur Tributino Menezes ; Helder Yudi Nagasse

Abstract

RNA processing is an important step in controlling gene expression in eukaryotic cells. Recent data from the literature have associated problems in pre-mRNA splicing with the development of different diseases, such as cancer and neurodegenerative diseases. In addition to mutations in sequences that encode important genes for these diseases, changes in proteins that bind to mRNAs can cause major phenotypic changes. More than 95% of human genes are composed of exons, coding sequences that remain in the mature mRNA, and introns, which are removed during splicing. In many cases, the same gene can generate different mRNAs through alternative splicing. Splicing is performed by the spliceosome, a large ribonucleoprotein machinery formed by 5 snRNAs and more than 100 proteins. Splicing regulation depends on the activity of RNA-binding proteins, responsible for mediating the interaction between RNAs and between RNAs and proteins. In this sense, splicing and the activity of regulatory proteins, such as hnRNPs, are important therapeutic targets. The control performed by these proteins can change cell phenotypes and promote characteristics that develop or not certain diseases. Thus, the hypothesis of this project is that generation of disease-related transcripts can be controlled by modulating the association of RNA-binding proteins to target sequences in pre-mRNAs. To investigate this hypothesis, we intend to design RNA oligos for the proteins hnRNP A1, hnRNP K, hnRNP M and TDP43, and verify the alternative splicing of genes important for tumor development, including genes that host oncogenic miRNAs. Furthermore, we intend to investigate whether the use of these oligos modulates the subcellular localization of proteins and cellular kinetics. Through analysis of RNA sequencing and mass spectrometry, we intend to investigate whether the use of these oligos can alter the function of proteins. The development of this project will allow to continue the research line on the regulation of splicing, including new tools as important therapeutic approaches to control RNA-associated diseases. It is expected that the results will also contribute to increase knowledge about basic mechanisms of activity of these proteins. (AU)