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A lentiviral vector with a short troponin-I promoter for tracking cardiomyocyte differentiation of human embryonic stem cells

Gene Therapy volume 15, pages 161–170 (2008)Cite this article

Abstract

Human embryonic stem cells (hESCs) may become important for cardiac repair due to their potentially unlimited ability to generate cardiomyocytes (CMCs). Moreover, genetic manipulation of hESC-derived CMCs would be a very promising technique for curing myocardial disorders. At the present time, however, inducing the differentiation of hESCs into CMCs is extremely difficult and, therefore, an easy and standardizable technique is needed to evaluate differentiation strategies. Vectors driving cardiac-specific expression may represent an important tool not only for monitoring new cardiac-differentiation strategies, but also for the manipulation of cardiac differentiation of ESCs. To this aim, we generated cardiac-specific lentiviral vectors (LVVs) in which expression is driven by a short fragment of the cardiac troponin-I proximal promoter (TNNI3) with a human cardiac α-actin enhancer, and tested its suitability in inducing tissue-specific gene expression and ability to track the CMC lineage during differentiation of ESCs. We determined that (1) TNNI3-LVVs efficiently drive cardiac-specific gene expression and mark the cardiomyogenic lineage in human and mouse ESC differentiation systems (2) the cardiac α-actin enhancer confers a further increase in gene-expression specificity of TNNI3-LVVs in hESCs. Although this technique may not be useful in tracking small numbers of cells, data suggested that TNNI3-based LVVs are a powerful tool for manipulating human ESCs and modifying hESC-derived CMCs.

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Acknowledgements

This work was supported by grants from the Italian Ministry of University and Research, the European Community (EU FP6 Grant LSHM-CT-2005-018833, EUGeneHeart) for the part related to lentiviral vector generation, and private funds to GC. We thank the technical help from Antonio Addario, Alessandra Benetello, Marco Santonastasi and Domenico Vignone and Angela Pitisci. We are grateful to Lucio Barile, Elisa Messina and Alessandro Giacomello for the use of cardiac progenitor cells.

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Authors and Affiliations

  1. Laboratory of Molecular Cardiology, San Raffaele Biomedical Science Park Foundation, Rome, Italy

    P Gallo, S Grimaldi, P Gallo & G Condorelli

  2. Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Multimedica, Milan, Italy

    P Gallo, M V G Latronico & G Condorelli

  3. Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore Sanità, Rome, Italy

    D Bonci, A Pagliuca & C Peschle

  4. Department of Biomedical Sciences, University of Padua, Padua, Italy

    S Ausoni

  5. Department of Medicine, Division of Cardiology, University of California, San Diego, CA, USA,

    G Condorelli

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  1. P Gallo
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Correspondence to G Condorelli.

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Supplementary Information accompanies the paper on Gene Therapy website (http://www.nature.com/gt)

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Gallo, P., Grimaldi, S., Latronico, M. et al. A lentiviral vector with a short troponin-I promoter for tracking cardiomyocyte differentiation of human embryonic stem cells. Gene Ther 15, 161–170 (2008). https://doi.org/10.1038/sj.gt.3303017

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  • DOI: https://doi.org/10.1038/sj.gt.3303017

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