Adenovirus-mediated gene transfer and expression of human beta-glucuronidase gene in the liver, spleen, and central nervous system in mucopolysaccharidosis type VII mice
Adenovirus-mediated gene transfer and expression of human beta-glucuronidase gene in the liver, spleen, and central nervous system in mucopolysaccharidosis type VII mice
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DOI：
10.1073/pnas.94.4.1287
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发表时间：
1997-02-18
影响因子：
11.1
通讯作者：
Eto, Y
中科院分区：
文献类型：
--
作者：
Ohashi, T;Watabe, K;Eto, Y
粘多糖沉积症VII型（Sly综合征）是由溶酶体酶β-葡萄糖醛酸苷酶的遗传性缺乏引起的溶酶体贮积病。这种疾病的小鼠模型已经得到很好的表征，并用于研究许多形式的实验疗法，包括基因疗法。我们生产了表达人β-葡萄糖醛酸糖苷酶的重组腺病毒，并将该重组腺病毒静脉内给予β-葡萄糖醛酸糖苷酶缺陷小鼠。第16天，肝脏和脾脏中的β-葡萄糖醛酸酶活性分别升高至杂合子酶水平的40%和20%。表达持续至少35天。这些组织的病理异常也得到改善，治疗小鼠的尿糖胺聚糖水平升高降低。然而，肾脏和大脑中的β-葡萄糖醛酸酶活性没有显著增加。将重组腺病毒直接注入突变小鼠的侧脑室后，粗制脑匀浆中的β-葡萄糖醛酸酶活性增加至杂合子活性的30%。脑组织中β-葡萄糖醛酸酶活性的组织化学显示，该酶活性主要在室管膜细胞和脉络膜中。然而，在某些区域，腺病毒介导的基因表达在与血管相关的脑实质和脑膜中也很明显。这些结果表明，腺病毒介导的基因传递可能会改善中枢神经系统病理粘多糖样变性，除了纠正内脏病理。
Mucopolysaccharidosis type VII (Sly syndrome) is a lysosomal storage disease caused by inherited deficiency of the lysosomal enzyme beta-glucuronidase. A murine model of this disorder has been well characterized and used to study a number of forms of experimental therapies, including gene therapy. We produced recombinant adenovirus that expresses human beta-glucuronidase and administered this recombinant adenovirus to beta-glucuronidase-deficient mice intravenously. The beta-glucuronidase activities in liver and spleen were elevated to 40% and 20%, respectively, of the heterozygote enzymatic level at day 16. Expression persisted for at least 35 days. Pathological abnormalities of these tissues were also improved, and the elevated levels of urinary glycosaminoglycans were reduced in treated mice. However, the beta-glucuronidase activity in kidney and brain was not significantly increased. After administration of the recombinant adenovirus directly into the lateral ventricles of mutant mice, the beta-glucuronidase activity in crude brain homogenates increased to 30% of heterozygote activity. Histochemical demonstration of beta-glucuronidase activity in brain revealed that the enzymatic activity was mainly in ependymal cells and choroid. However, in some regions, the adenovirus-mediated gene expression was also evident in brain parenchyma associated with vessels and in the meninges. These results suggest that adenovirus-mediated gene delivery might improve the central nervous system pathology of mucopolysaccharidosis in addition to correcting visceral pathology.