title: Defined MicroRNAs Induce Aspects of Maturation in Mouse and Human Embryonic-Stem-Cell-Derived Cardiomyocytes.
abstract: Pluripotent-cell-derived cardiomyocytes have great potential for use in research and medicine, but limitations in their maturity currently constrain their usefulness. Here, we report a method for improving features of maturation in murine and human embryonic-stem-cell-derived cardiomyocytes (m/hESC-CMs). We found that coculturing m/hESC-CMs with endothelial cells improves their maturity and upregulates several microRNAs. Delivering four of these microRNAs, miR-125b-5p, miR-199a-5p, miR-221, and miR-222 (miR-combo), to m/hESC-CMs resulted in improved sarcomere alignment and calcium handling, a more negative resting membrane potential, and increased expression of cardiomyocyte maturation markers. Although this could not fully phenocopy all adult cardiomyocyte characteristics, these effects persisted for two months following delivery of miR-combo. A luciferase assay demonstrated that all four miRNAs target ErbB4, and siRNA knockdown of ErbB4 partially recapitulated the effects of miR-combo. In summary, a combination of miRNAs induced via endothelial coculture improved ESC-CM maturity, in part through suppression of ErbB4 signaling.
authors.affiliation: Institute of Clinical Medicine, National Cheng Kung University, Tainan 704, Taiwan; Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Desy S
authors.initials: DS
authors.lastname: Lee
authors.affiliation: Department of Medicine, National Cheng Kung University, Tainan 704, Taiwan. Electronic address: jyhhong@mail.ncku.edu.tw.
authors.forename: Jyh-Hong
authors.initials: JH
authors.lastname: Chen
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan.
authors.forename: David J
authors.initials: DJ
authors.lastname: Lundy
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Chung-Hung
authors.initials: CH
authors.lastname: Liu
authors.affiliation: Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu 300, Taiwan.
authors.forename: Shiaw-Min
authors.initials: SM
authors.lastname: Hwang
authors.affiliation: Department of Pathology, Center for Cardiovascular Biology, Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA.
authors.forename: Lil
authors.initials: L
authors.lastname: Pabon
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Ru-Chi
authors.initials: RC
authors.lastname: Shieh
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Chien-Chang
authors.initials: CC
authors.lastname: Chen
authors.affiliation: Department of Physiology, National Cheng Kung University, Tainan 704, Taiwan.
authors.forename: Sheng-Nan
authors.initials: SN
authors.lastname: Wu
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Yu-Ting
authors.initials: YT
authors.lastname: Yan
authors.affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan.
authors.forename: Sho-Tone
authors.initials: ST
authors.lastname: Lee
authors.affiliation: Institute of Clinical Medicine, National Cheng Kung University, Tainan 704, Taiwan.
authors.forename: Po-Min
authors.initials: PM
authors.lastname: Chiang
authors.affiliation: Department of Bioengineering, University of California at San Diego, San Diego, CA 92093, USA.
authors.forename: Shu
authors.initials: S
authors.lastname: Chien
authors.affiliation: Department of Pathology, Center for Cardiovascular Biology, Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA; Department of Bioengineering, University of Washington, Seattle, WA 98109, USA; Department of Medicine/Cardiology, University of Washington, Seattle, WA 98109, USA.
authors.forename: Charles E
authors.initials: CE
authors.lastname: Murry
authors.affiliation: Institute of Clinical Medicine, National Cheng Kung University, Tainan 704, Taiwan; Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan; Department of Bioengineering, University of Washington, Seattle, WA 98109, USA; Division of Cardiovascular Surgery, National Taiwan University Hospital, Taipei 100, Taiwan. Electronic address: phsieh@ibms.sinica.edu.tw.
authors.forename: Patrick C H
authors.initials: PC
authors.lastname: Hsieh
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002450:Cell Communication; D002454:Cell Differentiation; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000648:ultrastructure; D042783:Endothelial Cells / Q000378:metabolism / Q000648:ultrastructure; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008564:Membrane Potentials; D051379:Mice; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure; D032383:Myocytes, Cardiac / Q000378:metabolism* / Q000648:ultrastructure; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D066247:Receptor, ErbB-4 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D012518:Sarcomeres / Q000378:metabolism / Q000648:ultrastructure; D015398:Signal Transduction
chemical_list: D015415:Biomarkers; C546122:MIRN221 microRNA, mouse; C546123:MIRN222 microRNA, mouse; D035683:MicroRNAs; C527468:Mirn125 microRNA, mouse; C534780:Mirn199 microRNA, mouse; D034741:RNA, Small Interfering; D008156:Luciferases; C587690:Erbb4 protein, mouse; D066247:Receptor, ErbB-4; D002118:Calcium