Reay D P, Bilbao R, Koppanati B M, Cai L, O'Day T L, Jiang Z, Zheng H, Watchko J F, Clemens P R
Department of Neurology, University of Pittsburgh, Pittsburgh, PA 15213, USA.
Gene Ther. 2008 Apr;15(7):531-6. doi: 10.1038/gt.2008.8. Epub 2008 Feb 14.
In utero gene therapy for genetic diseases, such as muscular dystrophies, offers potential advantages over postnatal treatment including vector delivery at the earliest point in the disease and treatment prior to full maturation of the immune system. This study examines in utero gene delivery of full-length murine dystrophin to the murine mdx model for Duchenne muscular dystrophy using a high-capacity adenoviral vector. We examined dystrophin expression, spread of vector, morphology and specific force production of the tibialis anterior muscle 9 weeks after intramuscular in utero injection. Recombinant dystrophin was expressed in the hindlimb muscles, with the majority of animals having expression in two muscles of the injected hindlimb. The dystrophin-glycoprotein complex was restored in those muscle fibers expressing recombinant dystrophin. Analysis of the percentage of dystrophin-expressing muscle fibers with centrally placed nuclei revealed effective protection from cycles of degeneration and regeneration normally seen in muscle fibers lacking dystrophin. However, due to low levels of muscle gene transfer, further advances in the efficiency of adenoviral vector-mediated gene delivery would be required for clinical applications of in utero gene therapy for primary myopathies such as Duchenne muscular dystrophy.
针对诸如肌营养不良症等遗传性疾病的宫内基因治疗，相较于出生后治疗具有潜在优势，包括在疾病最早阶段进行载体递送以及在免疫系统完全成熟之前进行治疗。本研究使用高容量腺病毒载体，研究将全长小鼠肌营养不良蛋白宫内基因递送至杜兴氏肌营养不良症的小鼠mdx模型。我们在宫内肌肉注射9周后，检测了胫骨前肌的肌营养不良蛋白表达、载体传播、形态以及比肌力产生情况。重组肌营养不良蛋白在后肢肌肉中表达，大多数动物在注射后肢的两块肌肉中都有表达。在表达重组肌营养不良蛋白的肌纤维中，肌营养不良蛋白-糖蛋白复合物得以恢复。对具有中央核的表达肌营养不良蛋白的肌纤维百分比进行分析发现，其能有效防止在缺乏肌营养不良蛋白的肌纤维中通常所见的退化和再生循环。然而，由于肌肉基因转移水平较低，对于杜兴氏肌营养不良症等原发性肌病的宫内基因治疗临床应用而言，腺病毒载体介导的基因递送效率还需要进一步提高。
