Successful compensation for dystrophin deficiency by a helper-dependent adenovirus expressing full-length utrophin
Successful compensation for dystrophin deficiency by a helper-dependent adenovirus expressing full-length utrophin
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DOI：
10.1038/sj.mt.6300260
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发表时间：
2007-10-01
影响因子：
12.4
通讯作者：
Karpati, George
中科院分区：
文献类型：
--
作者：
Deol, Jatinderpal R.;Danialou, Gawiyou;Karpati, George
辅助腺病毒载体（AdV）介导的全长肌营养不良蛋白表达可显著减轻mdx小鼠的营养不良表型。然而，肌营养不良蛋白作为一种新抗原，引发抗体的形成。作为一种替代方法，我们评估了全长小鼠肌营养不良蛋白的基因转移，这是一种功能同源的肌营养不良蛋白，通常只存在于神经肌肉交界处。在胫前肌（TA）中单次注射编码肌营养因子的辅助依赖腺病毒载体提供了非常好的转导，58%的新生儿纤维显示肌上皮肌营养因子表达，35%的成人纤维显示肌营养因子阳性（Utr(+)）。肌营养蛋白的存在阻止了新生儿的广泛坏死，阻止了成人的进一步坏死，并导致肌营养不良蛋白相关蛋白在注射后长达1年的肌上皮表达恢复。在新生儿和成人中均观察到明显的生理改善。体液反应和细胞免疫反应均未明显增加。然而，最初的高营养蛋白表达有一个时间相关的下降。尽管病毒DNA在新生期注射的动物体内持续存在，但成年小鼠肌肉中的病毒DNA水平下降。这些结果表明，虽然肌营养因子基因转移导致营养不良表型的改善，但在肌营养因子表达缺失时，这种效果并不持续。
Helper-dependent adenovirus vector (AdV)-mediated full-length dystrophin expression leads to significant mitigation of the dystrophic phenotype of the mdx mouse. However, dystrophin, as a neoantigen, elicits antibody formation. As an alternative approach, we evaluated gene transfer of full-length murine utrophin, a functional homologue of dystrophin that is normally present only at the neuromuscular junction. A single injection in the tibialis anterior (TA) muscle of the helper-dependent adenovirus vector encoding utrophin provided very good transduction, with 58% of fibers demonstrating sarcolemmal utrophin expression in the neonates, and 35% utrophin-positive (Utr(+)) fibers in adults. The presence of utrophin prevented extensive necrosis in the neonates, halted further necrosis in the adults, and led to restoration of sarcolemmal expression of dystrophin-associated proteins up to 1 year after injection. Marked physiological improvement was observed in both neonates and adults. Neither increased humoral responses nor cellular immune responses were evident. However, there was a time-related decline of the initial high utrophin expression. Although viral DNA persisted in animals that were injected in the neonatal stage, viral DNA levels decreased in muscles of adult mice. These results demonstrate that although utrophin gene transfer leads to amelioration of the dystrophic phenotype, the effects are not sustained upon loss of utrophin expression.