CHARACTERIZATION OF MOLECULAR MARKERS OF MATURATION IN HIPSC-CMS AS A MODEL FOR CARDIOREGENERATIVE THERAPIES. PRELIMINARY RESULTS

JULIA MARIA HALEK; JOAQUIN SMUCLER; MARIA AGUSTINA SCARAFIA; ALBERTO CROTTOGINI; MARIANO NICOLAS BELAICH; SANTIAGO GABRIEL MIRIUKA; ALEJANDRO DAMIÁN LA GRECA; PAOLA LOCATELLI

Mar del Plata

Congreso; LXVII Reunion Anual de Sociedades de Biociencias; 2022

Sociedad Argentina de Investigacion Clinica (SAIC)

Our objective was to characterize the molecular markers of adult cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) maintained in culture to induce temporal maturation, for its use as in vitro models in cardiac regeneration. Cardiovascular disease is the leading cause of morbidity and mortality worldwide. In this context, hiPSC-CMs represent a promising model for the design and study of therapeutic strategies. Considering its immature fetal phenotype and the lack of scientific consensus about the time in culture to consider them adult, it is necessary to generate and characterize hiPSC-CMs from long-term cultures. Objective: To characterize the temporal expression of molecular maturation markers to define the culture duration necessary to obtain hiPSC-CM with an adult phenotype, for its further use in the assessment of cardioregenerative gene therapies. Methodology: hiPSC were cultured and differentiated into cardiomyocytes. Samples were obtained at different times in culture (21, 40, 50, and 70 days), RNA was obtained and gene expression analysis of cardiomyocyte molecular markers was assessed through RT-qPCR. Results: as time in culture increased, we observed a rise in expression level of oxidative metabolism genes, such as PPARGC1A and COX6A1, and a decrease in the expression of cell cycle genes such as CDK1, CCND2, CCNA2, CCNB1 and AURKB. We also observed an increase in expression levels of RYR2 gene, which encodes the adult CM ryanodine receptor. Conclusion: The expression pattern of molecular markers in 70-day cultures of hiPSC-CMs is consistent with the adult cardiomyocyte phenotype. We conclude that 70-day cultures of hiPSC-CMs provide an appropriate model to study cardiac regeneration therapies in adult hearts.