Functional Involvements of Heterogeneous Nuclear Ribonucleoprotein A1 in Smooth Muscle Differentiation from Stem Cells In Vitro and In Vivo§

Authors

  • Yuan Huang,

    1. Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
    2. Department of Cardiology, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China
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  • Luyang Lin,

    1. Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
    2. Guangzhou Institute of Dermatology, Guangzhou, People's Republic of China
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  • Xiaotian Yu,

    1. Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
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  • Guanmei Wen,

    1. Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
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  • Xiangyuan Pu,

    1. Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
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  • Hanqing Zhao,

    1. Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
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  • Changcun Fang,

    1. Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
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  • Jianhua Zhu,

    1. Department of Cardiology, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China
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  • Shu Ye,

    1. Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
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  • Li Zhang,

    Corresponding author
    1. Department of Cardiology, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China
    • Department of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou 310003, Zhejiang, People's Republic of China
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    • Author contributions: Y.H., L.L., X.Y., G.W., X.P., H.Z., and C.F.: collection and/or assembly of data; J.Z.: provision of study material; S.Y.: manuscript writing and/or provision of study materials; L.Z.: provision of study materials and/or financial support; Q.X.: conception and design, financial support, administrative support, provision of study material, collection and/or assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript. Y.H. and L.L. contributed equally to this article.

    • Telephone:+86-571-87236794; Fax: +86-571-87236794

  • Qingzhong Xiao

    Corresponding author
    1. Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
    • Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and the London School of Medicine and Dentistry, John Vane Science Centre, Charterhouse Square, London EC1M 6BQ, U.K.
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    • Telephone: +44(0)2078828263; Fax: +44(0)2078823408


  • Disclosure of potential conflicts of interest is found at the end of this article.

  • §

    First published online in STEM CELLSEXPRESS January 17, 2013.

Abstract

To investigate the functional involvements of heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1) in smooth muscle cell (SMC) differentiation from stem cells, embryonic stem cells were cultivated on collagen IV-coated plates to allow for SMC differentiation. We found that hnRNPA1 gene and protein expression was upregulated significantly during differentiation and coexpressed with SMC differentiation markers in the stem cell-derived SMCs as well as embryonic SMCs of 12.5 days of mouse embryos. hnRNPA1 knockdown resulted in downregulation of smooth muscle markers and transcription factors, while enforced expression of hnRNPA1 enhanced the expression of these genes. Importantly, knockdown of hnRNPA1 also resulted in impairment of SMC differentiation in vivo. Moreover, we demonstrated that hnRNPA1 could transcriptionally regulate SMC gene expression through direct binding to promoters of Acta2 and Tagln genes using luciferase and chromatin immunoprecipitation assays. We further demonstrated that the binding sites for serum response factor (SRF), a well-investigated SMC transcription factor, within the promoter region of the Acta2 and Tagln genes were responsible for hnRNPA1-mediated Acta2 and Tagln gene expression using in vitro site-specific mutagenesis and luciferase activity analyses. Finally, we also demonstrated that hnRNPA1 upregulated the expression of SRF, myocyte-specific enhancer factor 2c (MEF2c), and myocardin through transcriptional activation and direct binding to promoters of the SRF, MEF2c, and Myocd genes. Our findings demonstrated that hnRNPA1 plays a functional role in SMC differentiation from stem cells in vitro and in vivo. This indicates that hnRNPA1 is a potential modulating target for deriving SMCs from stem cells and cardiovascular regenerative medicine. STEM CELLS 2013;31:906–917

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