title: Timely inhibition of Notch signaling by DAPT promotes cardiac differentiation of murine pluripotent stem cells.
abstract: The Notch signaling pathway plays versatile roles during heart development. However, there is contradictory evidence that Notch pathway either facilitates or impairs cardiomyogenesis in vitro. In this study, we developed iPSCs by reprogramming of murine fibroblasts with GFP expression governed by Oct4 promoter, and identified an effective strategy to enhance cardiac differentiation through timely modulation of Notch signaling. The Notch inhibitor DAPT (N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester) alone drove the iPSCs to a neuronal fate. After mesoderm induction of embryoid bodies initiated by ascorbic acid (AA), the subsequent treatment of DAPT accelerated the generation of spontaneously beating cardiomyocytes. The timed synergy of AA and DAPT yielded an optimal efficiency of cardiac differentiation. Mechanistic studies showed that Notch pathway plays a biphasic role in cardiomyogenesis. It favors the early-stage cardiac differentiation, but exerts negative effects on the late-stage differentiation. Therefore, DAPT administration at the late stage enforced the inhibition of endogenous Notch activity, thereby enhancing cardiomyogenesis. In parallel, DAPT dramatically augmented the expression of Wnt3a, Wnt11, BMP2, and BMP4. In conclusion, our results highlight a practicable approach to generate cardiomyocytes from iPSCs based on the stage-specific biphasic roles of Notch signaling in cardiomyogenesis.
authors.affiliation: Stem Cell Research Center, Department of Cell Biology, School of Basic Medical Sciences, Peking University, Haidian District, Beijing, China.
authors.forename: Yinan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Stem Cell Research Center, Department of Cell Biology, School of Basic Medical Sciences, Peking University, Haidian District, Beijing, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Peking University, Haidian District, Beijing, China.
authors.forename: Kaiyu
authors.initials: K
authors.lastname: Liu
authors.affiliation: Center of Medical and Health Analysis, School of Basic Medical Sciences, Peking University, Haidian District, Beijing, China.
authors.forename: Qihua
authors.initials: Q
authors.lastname: He
authors.affiliation: Stem Cell Research Center, Department of Cell Biology, School of Basic Medical Sciences, Peking University, Haidian District, Beijing, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Han
authors.affiliation: Department of histology and embryology, Institute of Chinese Medicine, Hunan University of Chinese Medicine, Science Garden District of Hanpu, Changsha, Hunan, China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Sun
authors.affiliation: Department of Biology, College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua, Zhejiang, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Li
authors.affiliation: Stem Cell Research Center, Department of Cell Biology, School of Basic Medical Sciences, Peking University, Haidian District, Beijing, China.
authors.forename: Li
authors.initials: L
authors.lastname: Shen
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D004151:Dipeptides / Q000494:pharmacology*; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000502:physiology* / Q000637:transplantation; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D008822:Mice, Transgenic; D032386:Myoblasts, Cardiac / Q000378:metabolism*; D051881:Receptor, Notch1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction; D013724:Teratoma / Q000209:etiology / Q000473:pathology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D004151:Dipeptides; C419410:N-(N-(3,5-difluorophenacetyl)alanyl)phenylglycine tert-butyl ester; C497762:Notch1 protein, mouse; D051881:Receptor, Notch1; D014157:Transcription Factors