The role of NLRP3 inflammasome in models of cardiomyocytic hypertrophy. Use of human cardiomyocytes as an experimental model.

• Grant number: 23/08673-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2024
• End date: December 31, 2025
• Field of knowledge: Biological Sciences - Physiology - Physiology of Organs and Systems
• Principal Investigator: Maria Luiza de Morais Barreto de Chaves
• Grantee: Eloisa Olher Gomes
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 19/17031-2 - Inflammasome in the cardiac physiopathology, AP.TEM

Abstract

According to Worlds Health Organization, cardiovascular diseases are the main cause of death worldwide, as it reinforces the importance of studies that lead to a greater understanding of the mechanisms by which these diseases are installed and developed. Among them, there Heart Failure (HF), an epidemic disease with high rates of morbidity and mortality. HF is commonly preceded by cardiac hypertrophy (CH), characterized by an increase in cardiac mass at the expense of the addition of contractile fibers that can be classified in two categories: the physiological hypertrophy and the pathological hypertrophy, the latter being irreversible and capable of progressing to HF. Several stimuli can lead to CH, including the inflammatory process, with the NLRP3 inflammasome being an important regulator of this process. Due to the limitations in the use of primary cardiomyocytes, cell lines were developed for molecular and genetic studies, among them the AC16 line. These cells come from adult human ventricular cardiomyocytes fused with fibroblasts transfected with the SV40 oncogene. In this scenario, the object of this project is to realize the characterization of the AC16 cell line and its response to different hypertrophic stimuli, in order to analyze the participation of the NRLP3 inflammasome in the pathogenesis of cardiomyocyte hypertrophy in the future. For this, the cells will be thawed and their growth curve will be made, as well as the characterization of the phenotype by immunocytochemistry with markers of cardiomyocytes and fibroblasts. The cells will be stimulated with T3 and Isoproterenol to induce physiological and pathological hypertrophy respectively with dose x time x response curves. Total RNA and total proteins will be extracted from these cultures for analysis by RT-PCR and western blot of gene and protein expression of NLRP3 inflammasome components (NLRP3, ASC, and Caspase I).