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Fetal muscle gene transfer is not enhanced by an RGD capsid modification to high-capacity adenoviral vectors

Gene Therapy volume 10, pages 1821–1829 (2003)Cite this article

Abstract

High levels of αv integrin expression by fetal muscle suggested that vector re-targeting to integrins could enhance adenoviral vector-mediated transduction, thereby increasing safety and efficacy of muscle gene transfer in utero. High-capacity adenoviral (HC-Ad) vectors modified by an Arg-Gly-Asp (RGD) peptide motif in the HI loop of the adenoviral fiber (RGD-HC-Ad) have demonstrated efficient gene transfer through binding to αv integrins. To test integrin targeting of HC-Ad vectors for fetal muscle gene transfer, we compared unmodified and RGD-modified HC-Ad vectors. In vivo, unmodified HC-Ad vector transduced fetal mouse muscle with four-fold higher efficiency compared to RGD-HC-Ad vector. Confirming that the difference was due to muscle cell autonomous factors and not mechanical barriers, transduction of primary myogenic cells isolated from murine fetal muscle in vitro demonstrated a three-fold better transduction by HC-Ad vector than by RGD-HC-Ad vector. We hypothesized that the high expression level of coxsackievirus and adenovirus receptor (CAR), demonstrated in fetal muscle cells both in vitro and in vivo, was the crucial variable influencing the relative transduction efficiencies of HC-Ad and RGD-HC-Ad vectors. To explore this further, we studied transduction by HC-Ad and RGD-HC-Ad vectors in paired cell lines that expressed αv integrins and differed only by the presence or absence of CAR expression. The results increase our understanding of factors that will be important for retargeting HC-Ad vectors to enhance gene transfer to fetal muscle.

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Acknowledgements

We thank Steve K Wendell (University of Pittsburgh, Pittsburgh, PA, USA) for advice on real-time PCR. We thank Martin P Ontell and Charles Washabaugh (University of Pittsburgh, Pittsburgh, PA, USA) and Susan Schachtner (Children's Hospital of Philadelphia, Philadelphia, PA, USA) for instruction on fetal injections. We thank David Fink and Marina Mata (University of Pittsburgh, Pittsburgh, PA, USA) for technical support. This work was supported by NIH Grant P01 AR45925 (PRC), Muscular Dystrophy Association grants (PRC), and a postdoctoral fellowship grant from the Fundación Ramón Areces (RB).

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Authors and Affiliations

  1. Department of Neurology, University of Pittsburgh, Pittsburgh, PA, USA

    R Bilbao, D P Reay, T Hughes, L Goldberg & P R Clemens

  2. Center for Molecular Medicine, University of Cologne, Cologne, Germany

    V Biermann, C Volpers & S Kochanek

  3. Division of Immunologic and Infectious Diseases, The Children's Hospital of Philadelphia, Philadelphia, PA, USA

    J Bergelson

  4. Department of Veterans Affairs Medical Center, Neurology Service, Pittsburgh, PA, USA

    P R Clemens

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  1. R Bilbao
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Bilbao, R., Reay, D., Hughes, T. et al. Fetal muscle gene transfer is not enhanced by an RGD capsid modification to high-capacity adenoviral vectors. Gene Ther 10, 1821–1829 (2003). https://doi.org/10.1038/sj.gt.3302084

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