Neuregulin/ErbB Signaling Regulates Cardiac Subtype Specification in Differentiating Human Embryonic Stem Cells

• Published in: Circulation research Vol. 107; no. 6; pp. 776 - 786
• Main Authors: Zhu, Wei-Zhong, Xie, Yiheng, Moyes, Kara White, Gold, Joseph D, Askari, Bardia, Laflamme, Michael A
• Format: Journal Article
• Language: English
• Published: Hagerstown, MD American Heart Association, Inc 17.09.2010; Lippincott Williams & Wilkins
• Subjects: Animals; Biological and medical sciences; Cell Differentiation - physiology; Cell Line; Cells, Cultured; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Embryonic Stem Cells - physiology; Fundamental and applied biological sciences. Psychology; Humans; Mice; Myocytes, Cardiac - classification; Myocytes, Cardiac - cytology; Myocytes, Cardiac - metabolism; Neuregulin-1 - physiology; Receptor, Epidermal Growth Factor - physiology; Signal Transduction - physiology; Sinoatrial Node - cytology; Sinoatrial Node - embryology; Sinoatrial Node - metabolism; Vertebrates: cardiovascular system; Human; Signal transduction; Vertebrata; Embryo; Mammalia; embryonic stem cell; Pacemaker; Stem cell; Electrophysiology; Neuregulin; Circulatory system; Subtype
• ISSN: 0009-7330; 1524-4571
• DOI: 10.1161/CIRCRESAHA.110.223917

RATIONALE:Human embryonic stem cell–derived cardiomyocytes (hESC-CMs) exhibit either a “working” chamber or a nodal-like phenotype. To generate optimal hESC-CM preparations for eventual clinical application in cell-based therapies, we will need to control their differentiation into these specialized cardiac subtypes. OBJECTIVE:To demonstrate intact neuregulin (NRG)-1β/ErbB signaling in hESC-CMs and test the hypothesis that this signaling pathway regulates cardiac subtype abundance in hESC-CM cultures. METHODS AND RESULTS:All experiments used hESC-CM cultures generated using our recently reported directed differentiation protocol. To support subsequent action potential phenotyping approaches and provide a higher-throughput method of determining cardiac subtype, we first developed and validated a novel genetic label that identifies nodal-type hESC-CMs. Next, control hESC-CM preparations were compared to those differentiated in the presence of exogenous NRG-1β, an anti–NRG-1β neutralizing antibody, or the ErbB antagonist AG1478. We used 3 independent approaches to determine the ratio of cardiac subtypes in the resultant populationsdirect action potential phenotyping under current-clamp, activation of the aforementioned genetic label, and subtype-specific marker expression by RT-PCR. Using all 3 end points, we found that inhibition of NRG-1β/ErbB signaling greatly enhanced the proportion of cells showing the nodal phenotype. CONCLUSIONS:NRG-1β/ErbB signaling regulates the ratio of nodal- to working-type cells in differentiating hESC-CM cultures and presumably functions similarly during early human heart development. We speculate that, by manipulating NRG-1β/ErbB signaling, it will be possible to generate preparations of enriched working-type myocytes for infarct repair, or, conversely, nodal cells for potential use in a biological pacemaker.