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

Citation
A. Baranov et al., Local and distant transfection of mdx muscle fibers with dystrophin and LacZ genes delivered in vivo by synthetic microspheres, GENE THER, 6(8), 1999, pp. 1406-1414
Citations number
22
Categorie Soggetti
Molecular Biology & Genetics
Journal title
GENE THERAPY
ISSN journal
09697128 → ACNP
Volume
6
Issue
8
Year of publication
1999
Pages
1406 - 1414
Database
ISI
SICI code
0969-7128(199908)6:8<1406:LADTOM>2.0.ZU;2-P
Abstract
Patterns of dystrophin and beta-galactosidase expression were examined in m dx mice after i.m. injections of synthetic microspheres (MF-2) loaded with full-length (pHSADy) or mini-dystrophin gene (pSG5dys) cDNA plasmid constru cts or with LacZ marker gene (pCMV-LacZ). A single injection of 25 mu g pHS ADy 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 e xposure compared with only 0.6% DPM in intact (non-injected) mdx mice. A hi gh proportion of DPM (17.6% and 10.8%, respectively) was registered in both injected and contralateral muscles after mini: gene cDNA administration. M F-2/dystrophin cDNA particles 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 i n all skeletal muscles and in different infernal organs was proven by PCR a nd RT-PCR analysis. Patches of beta-galactosidase expression were abundant in injected muscle, and frequent in the contralateral and other skeletal mu scles as well as in diaphragm, heart and lungs. High levels of dystrophin c DNA expression, and an efficient distant transfection effect with preferent ial intranuclei inclusion of MF-2 vehicle, are very encouraging for the dev elopment of a new constructive strategy in gene therapy trials of DMD.