Optimization of total RNA extraction and amplification for miRNA analysis in patients with Familial Hypercholesterolemia

• Grant number: 23/11851-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): September 01, 2023
• Effective date (End): July 31, 2024
• Field of knowledge: Health Sciences - Pharmacy
• Principal Investigator: Mario Hiroyuki Hirata
• Grantee: Isadora Alves Damasceno
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 16/12899-6 - Genomics, epigenomics and pharmacogenomics characterization of familial hypercholesterolemia in the Brazilian population, AP.TEM

Abstract

Familial hypercholesterolemia (FH) is an autosomal dominant disease that affects cholesterol metabolism, causing high plasma concentrations of LDL particles. The genetic mechanisms underlying this condition are not fully understood. Recent studies have explored the importance of epigenomic mechanisms, including regulation by microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) in cholesterol metabolism. lncRNAs can influence gene transcription through several mechanisms, including epigenetic alterations. These molecules have also been associated with several human diseases, particularly cardiovascular disease. In this context, lncRNAs not only play a role in cholesterol levels, but also in the development of cardiovascular diseases. The proposed study focuses on the standardization of total RNA extraction for amplification and analysis of lncRNA differential expression in plasma from patients with FH. The objective is to analyze the expression of miR-552-3p, miR-499 and miR-941. The project will be submitted to relevant ethics committees, and participants will provide informed consent. Techniques used for analysis will include RNA extraction, spectrophotometry and RT-qPCR. In summary, understanding epigenetic mechanisms, especially involving lncRNAs, may better elucidate the pathophysiological processes of FH and offer new approaches for the prevention and treatment of associated cardiovascular diseases.