Cited4 is related to cardiogenic induction and maintenance of proliferation capacity of embryonic stem cell-derived cardiomyocytes during in vitro cardiogenesis

• Published in: PloS one Vol. 12; no. 8; p. e0183225
• Main Authors: Miake, Junichiro, Notsu, Tomomi, Higaki, Katsumi, Hidaka, Kyoko, Morisaki, Takayuki, Yamamoto, Kazuhiro, Hisatome, Ichiro
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 17.08.2017; Public Library of Science (PLoS)
• Subjects: Animals; Biology and Life Sciences; Calcium-binding protein; Cardiomyocytes; Cell division; Cell Proliferation; Cell Separation; Cells (biology); Cells, Cultured; Cloning; CREB-binding protein; Cyclic AMP response element-binding protein; Cytometry; Differentiation; Embryo cells; Embryonic stem cells; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Event-related potentials; Flow Cytometry; Gene expression; Gene Expression Regulation, Developmental; Heart; Heart - embryology; Heart cells; Heart diseases; Hypertrophy; In vitro methods and tests; Kinases; Medicine; Mice; Myocytes, Cardiac - cytology; Myocytes, Cardiac - metabolism; Polymerase chain reaction; Progenitor cells; Proteins; Regeneration; Research and Analysis Methods; RNA-directed DNA polymerase; Stem cell transplantation; Stem cells; Transcription; Transcription Factors - genetics; Transcription Factors - metabolism; Troponin; Troponin T; United States > US
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0183225

Cardiac progenitor cells have a limited proliferative capacity. The CREB-binding protein/p300-interacting transactivator, with the Glu/Asp-rich carboxy-terminal domain (Cited) gene family, regulates gene transcription. Increased expression of the Cited4 gene in an adult mouse is associated with exercise-induced cardiomyocyte hypertrophy and proliferation. However, the expression patterns and functional roles of the Cited4 gene during cardiogenesis are largely unknown. Therefore, in the present study, we investigated the expression patterns and functional roles of the Cited4 gene during in vitro cardiogenesis. Using embryoid bodies formed from mouse embryonic stem cells, we evaluated the expression patterns of the Cited4 gene by quantitative reverse transcriptase-polymerase chain reaction. Cited4 gene expression levels increased and decreased during the early and late phases of cardiogenesis, respectively. Moreover, Cited4 gene levels were significantly high in the cardiac progenitor cell population. A functional assay of the Cited4 gene in cardiac progenitor cells using flow cytometry indicated that overexpression of the Cited4 gene significantly increased the cardiac progenitor cell population compared with the control and knockdown groups. A cell proliferation assay, with 5-ethynyl-2'-deoxyuridine incorporation and Ki67 expression during the late phase of cardiogenesis, indicated that the number of troponin T-positive embryonic stem cell-direived cardiomyocytes with proliferative capacity was significantly greater in the overexpression group than in the control and knockdown groups. Our study results suggest that the Cited4 gene is related to cardiac differentiation and maintenance of proliferation capacity of embryonic stem cell-derived cardiomyocytes during in vitro cardiogenesis. Therefore, manipulation of Cited4 gene expression may be of great interest for cardiac regeneration.