Cardiomyogenic differentiation is fine-tuned by differential mRNA association with polysomes

Isabela Tiemy Pereira,Lucia Spangenberg,Rocío Amorín,Bruno Dallagiovanna,Hugo Naya,Marco Augusto Stimamiglio,Anny Waloski Robert

doi:10.1186/s12864-019-5550-3

Isabela Tiemy Pereira, Lucia Spangenberg + Show 5 more

Open Access

https://doi.org/10.1186/s12864-019-5550-3

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Abstract

BackgroundCardiac cell fate specification occurs through progressive steps, and its gene expression regulation features are still being defined. There has been an increasing interest in understanding the coordination between transcription and post-transcriptional regulation during the differentiation processes. Here, we took advantage of the polysome profiling technique to isolate and high-throughput sequence ribosome-free and polysome-bound RNAs during cardiomyogenesis.ResultsWe showed that polysome-bound RNAs exhibit the cardiomyogenic commitment gene expression and that mesoderm-to-cardiac progenitor stages are strongly regulated. Additionally, we compared ribosome-free and polysome-bound RNAs and found that the post-transcriptional regulation vastly contributes to cardiac phenotype determination, including RNA recruitment to and dissociation from ribosomes. Moreover, we found that protein synthesis is decreased in cardiomyocytes compared to human embryonic stem-cells (hESCs), possibly due to the down-regulation of translation-related genes.ConclusionsOur data provided a powerful tool to investigate genes potentially controlled by post-transcriptional mechanisms during the cardiac differentiation of hESC. This work could prospect fundamental tools to develop new therapy and research approaches.

Highlights

Cardiac cell fate specification occurs through progressive steps, and its gene expression regulation features are still being defined
Polysome profiling during human embryonic stem-cells (hESCs) cardiomyogenic differentiation The hES-NKX2–5eGFP/w reporter human embryonic stem cell line [27] was used to derive cardiomyocytes using a developmentally staged protocol [2, 28] that includes the induction of a cardiac mesoderm population on days 3 and 4 and a NKX2–5+/cTNT+ population by day 15 (Fig. 1a and b)
We found that the translational activity is reduced during differentiation; this became more evident when comparing hESC on D0 to cardiomyocytes on day 15 (D15)

Summary

Introduction

Cardiac cell fate specification occurs through progressive steps, and its gene expression regulation features are still being defined. There has been an increasing interest in understanding the coordination between transcription and post-transcriptional regulation during the differentiation processes. Cardiomyocytes have been derived from hESCs as an in vitro model to study cardiomyogenesis and as an attempt to produce clinically relevant cell populations [2,3,4,5]. Modeling congenital abnormalities of the heart or testing the cardiac toxicity of new drugs strengthen a particular interest in deriving cardiovascular lineages from pluripotent stem cells [2]. Large-scale studies help to uncover the complex and dynamic multi-layered regulation involving chromatin modifications, transcriptional, and post-transcriptional networks that control the reconfiguration of ESC gene expression program when

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