Transcriptome-guided functional analyses reveal novel biological properties and regulatory hierarchy of human embryonic stem cell-derived ventricular cardiomyocytes crucial for maturation

• Published in: PloS one 2013-10, Vol.8 (10), p.e77784-e77784
• Main Authors: Poon, Ellen, Yan, Bin, Zhang, Shaohong, Rushing, Stephanie, Keung, Wendy, Ren, Lihuan, Lieu, Deborah K, Geng, Lin, Kong, Chi-Wing, Wang, Jiaxian, Wong, Hau San, Boheler, Kenneth R, Li, Ronald A
• Format: Article
• Language: English
• Subjects: Adult; Animals; Apoptosis; Biological properties; Biomarkers - metabolism; Biomedical materials; Calcium; Calcium homeostasis; Calcium metabolism; Calcium transport; Cardiomyocytes; Cell cycle; Cell Differentiation; Cell migration; Cell Proliferation; Combinatorial analysis; Computer science; Consortia; Contractility; Contraction; DNA binding proteins; Electrophysiology; Embryo cells; Embryonic stem cells; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Fetuses; Fibroblasts; Gene expression; Gene Expression Profiling; Gene Expression Regulation, Developmental; Gene Regulatory Networks; Genes; Heart cells; Heart failure; Heart Ventricles - embryology; Heart Ventricles - metabolism; Homeostasis; Humans; Hydroxyapatite; Hypoxia; In vitro methods and tests; Ion transport; Kinases; Maturation; Medicine; Metabolism; Mice; Migratory species; Myocytes, Cardiac - cytology; Myocytes, Cardiac - metabolism; Nkx2.5 protein; Oligonucleotide Array Sequence Analysis; Oxygen; Phosphorylation; Physiology; Potassium; Promoter Regions, Genetic; Reactive oxygen species; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; Rodents; Stem cells; Transcription factors; Ventricle
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0077784

Human (h) embryonic stem cells (ESC) represent an unlimited source of cardiomyocytes (CMs); however, these differentiated cells are immature. Thus far, gene profiling studies have been performed with non-purified or non-chamber specific CMs. Here we took a combinatorial approach of using systems biology to guide functional discoveries of novel biological properties of purified hESC-derived ventricular (V) CMs. We profiled the transcriptomes of hESCs, hESC-, fetal (hF) and adult (hA) VCMs, and showed that hESC-VCMs displayed a unique transcriptomic signature. Not only did a detailed comparison between hESC-VCMs and hF-VCMs confirm known expression changes in metabolic and contractile genes, it further revealed novel differences in genes associated with reactive oxygen species (ROS) metabolism, migration and cell cycle, as well as potassium and calcium ion transport. Following these guides, we functionally confirmed that hESC-VCMs expressed IKATP with immature properties, and were accordingly vulnerable to hypoxia/reoxygenation-induced apoptosis. For mechanistic insights, our coexpression and promoter analyses uncovered a novel transcriptional hierarchy involving select transcription factors (GATA4, HAND1, NKX2.5, PPARGC1A and TCF8), and genes involved in contraction, calcium homeostasis and metabolism. These data highlight novel expression and functional differences between hESC-VCMs and their fetal counterparts, and offer insights into the underlying cell developmental state. These findings may lead to mechanism-based methods for in vitro driven maturation.