Characterization of lncRNAs expression profiles in carciomyocytes derived from pluripotent stem cells

MARIA AGUSTINA SCARAFIA; NATALIA LUCÍA SANTÍN VELAZQUE; ALAN MIQUEAS MÖBBS; GABRIEL NEIMAN; CAROLINA COLLI; ARIEL WAISMAN; LUCIA NATALIA MORO; CARLOS DANIEL LUZZANI; GUSTAVO EMILIO SEVLEVER; ALEJANDRO DAMIÁN LA GRECA; SANTIAGO GABRIEL MIRIUKA

Mar del Plata

Congreso; LXIII Annual Meeting of the Argentine Society for Clinical Investigation (SAIC); 2018

SAIC - Sociedad Argentina de Investigacion Clinica

Differentiation of human induced pluripotent stem cells (iPSC) into cardiomyocytes (CMs) is a resourceful approach for clinical and biological studies. Gaining insight into gene regulatory networks (GRN) that govern cardiac differentiation will allow a better understanding of cardiac development and commitment. Over the past ten years, long non-coding RNAs (lncRNAs), a heterogeneous group of non-coding transcripts, have gained importance as key modulators of these networks by fine tuning epigenetic, transcriptional and post-transcriptional expression, although their precise molecular mechanisms are not fully clarified.This work focused on characterizing the expression profiles of lncRNAs along cardiac differentiation by high throughput RNA next generation sequencing. CMs were generated from an iPSC line (FN2.1) developed in our laboratory by implementation and optimization of a previously reported protocol. Three cellular populations were studied: day 0 (pluripotent state); day 3.5 (early mesoderm progenitor, isolated by cell sorting); day 21 (immature cardiomyocyte, isolated by enrichment with selection medium - >80% CMs). Population identity was confirmed by qRT-PCR of specific gene markers and long non-coding markers previously reported in the literature. High-quality, total RNA was purified from three independent experiments on the three populations mentioned before and subjected to deep sequencing (~50 million 100bp-long pair-end reads) on Illumina platform. Initial assessment of data revealed homogeneous FPKM distribution and high interreplicate correlation indices (d0: r=0.90; d3.5: r=0.9; d21: r=0.85; p