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Local and distant transfection of mdx muscle fibers with dystrophin and LacZ genes delivered in vivo by synthetic microspheres

Gene Therapy volume 6, pages 1406–1414 (1999)Cite this article

Abstract

Patterns of dystrophin and β-galactosidase expression were examined in mdx mice after i.m. injections of synthetic microspheres (MF-2) loaded with full-length (pHSADy) or mini-dystrophin gene (pSG5dys) cDNA plasmid constructs or with LacZ marker gene (pCMV-LacZ). A single injection of 25 μg pHSADy into quadriceps femoris muscle resulted in 6.8% of dystrophin positive myofibers (DPM) in a given muscle; 8.4% of DPM in glutaeus muscle and 4.3% of DPM in quadriceps femoris muscle of contralateral limb on day 21 after exposure compared with only 0.6% DPM in intact (non-injected) mdx mice. A high proportion of DPM (17.6% and 10.8%, respectively) was registered in both injected and contralateral muscles after mini- gene cDNA administration. MF-2/dystrophin cDNA par- ticles were detected by FISH analysis in about 60–70% of myofiber nuclei in muscles of injected and contralateral limbs 7 days after application. The presence of human dystrophin cDNA and its products in all skeletal muscles and in different internal organs was proven by PCR and RT-PCR analysis. Patches of β-galactosidase expression were abundant in injected muscle, and frequent in the contralateral and other skeletal muscles as well as in diaphragm, heart and lungs. High levels of dystrophin cDNA expression, and an efficient distant transfection effect with preferential intranuclei inclusion of MF-2 vehicle, are very encouraging for the development of a new constructive strategy in gene therapy trials of DMD.

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Acknowledgements

The authors are grateful to ‘Biosystem XXI, St Petersburg, Russia for the original polymeric vehicle, to Dr Caroline Sewry and Dr Peter Strong (Hammersmith Hospital, London, UK) for the generous gift of P6 antibodies, Professor G Dickson (Royal Holloway University, London, UK) and Dr F Mavilio (TIGET, Milano, Italy) for dystrophin cDNA constructs. Personal gratitude to Professor T Partridge (Hammersmith Hospital, London, UK) for the stock of mdx mice, as well as permanent interest in this work and helpful constructive comments during manuscript preparation. This work was partly supported by the Russian State Program ‘Human Genome’ (grant 48/95).

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Authors and Affiliations

  1. Ott’s Institute of Obstetrics and Gynecology, St Petersburg, Russia

    A Baranov, P Glazkov, A Kiselev, O Ostapenko, T Ivaschenko & V Baranov

  2. Institute of Cytology, St Petersburg, Russia

    V Mikhailov

  3. Biosystem XXI Ltd, St Petersburg, Russia

    V Sabetsky

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  1. A Baranov
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  2. P Glazkov
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Baranov, A., Glazkov, P., Kiselev, A. et al. Local and distant transfection of mdx muscle fibers with dystrophin and LacZ genes delivered in vivo by synthetic microspheres. Gene Ther 6, 1406–1414 (1999). https://doi.org/10.1038/sj.gt.3300954

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