Screening of ovine cardiomyocyte cell cycle regulators as possible targets for cardiac regeneration

LÓPEZ, AYELÉN EMILCE; BAUZÁ, MARÍA DEL ROSARIO; CASTILLO, MARTHA GIOVANNA; GHIRINGHELLI, DANIEL; CERRUDO, CAROLINA; GIMÉNEZ, CARLOS SEBASTIÁN; CROTTOGINI, ALBERTO; BELAICH, MARIANO NICOLÁS; URANGA VEGA, MARTÍN; OLEA, FERNANDA DANIELA; LOCATELLI, PAOLA

Mar del Plata

Congreso; Reunión anual de la Sociedad Argentina de Fisiología; 2018

SAIC-SAI-SAFIS

Considering the poor results of clinical trials on cardiac regeneration using stem cells, there is a growing interest in developing gene therapies aimed at inducing the adult cardiomyocyte (CM) mitosis, an approach that would warrant efficient electromechanical coupling of new cells to the myocardial sincitium. Hence, disclosing novel CM cell cycle mediators that could be targeted with these therapies is essential. Thus, our objective was to analyze the ovine CM transcriptome and compare the expression levels of cell cycle regulator?s mRNAs in fetal sheep at two time points of gestation [90 days, when the CM is still mitotic (F90), and 130 days, when the CM has exited the cell cycle (F130)] and adults (A).Fetal (F90, n=3 and F130, n=3) and A (n=3) sheep were euthanized and samples from the left ventricle were collected. Total RNA isolation was performed and mRNA was purified. Samples were delivered to Macrogen Inc. for cDNA library construction and Next Generation Sequencing. Raw RNAseq reads of the three experimental groups were quantified and classified using bioinformatic tools according to expression levels and proposed function. Several genes related to cell cycle regulation showing significant expression differences were selected and their expression levels re-evaluated by Real Time PCR. For statistical analysis we used ANOVA-Bonferroni (p