Gene Therapy for Lysosomal Strage Disease

溶酶体疾病的基因治疗

基本信息

批准号：
07670905
负责人：
OHASHI Touya
金额：
$ 1.34万
依托单位：
The Jikei University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

Mucopolysaccharidosis type VII (Sly disease) 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. In this study, we generated recombinant adenovirus which express human beta-glucuronidase and administered this recombinant adenovirus to model mice intravenously. The beta-glucuronidase activities in liver and spleen were elevated and expression persisted for at least 35 days. Pathological adnormalities of these tissues were improved. Following administration of the recombinant adenovirus directly into the lateral ventricles of mutant mice, the recombinant adenovirus infected mainly ependymal cells and choroid. Only a small number of brain parenchymal cells were infected.As an alternative approach, we also study transfer of the human beta-glucuronidase gene to human hematopoietic cells. Bone marrow transplantation in mucopolysaccharidosis patients in early clinical stages has been reported to halt the progression of neurological deterioration. As a first step, we generated recombinant retrovirus which express human beta-glucuronidase gene and infected this virus to CD34+ cells purified from umbilical cord blood. Using a centrifugation promoted infection protocol, we got transduction efficiency (Ave.67%) in colony forming unit granulocyte/macrophage and burst forming unit erythrocyte. Experiments are underway whether transduced CD34+ cells migrate to brain in severe combined immuno-deficient mice.

粘多糖型VII（狡猾的疾病）是一种溶酶体储存疾病，由溶酶体酶β-葡萄糖醛酸酶的遗传缺乏引起。该疾病的鼠模型已得到很好的特征，并用于研究包括基因疗法在内的多种实验疗法。在这项研究中，我们产生了重组腺病毒，该腺病毒表达人β-葡萄糖醛酸酶，并给予该重组腺病毒以静脉内对小鼠进行建模。肝脏和脾脏中的β-葡萄糖醛酸酶活性升高，表达持续至少35天。这些组织的病理性依赖性得到了改善。将重组腺病毒直接施用到突变小鼠外侧心室后，重组腺病毒主要感染了主要des膜细胞和脉络膜。仅感染了少数脑实质细胞。作为另一种方法，我们还研究了人β-葡萄糖醛酸糖苷酶基因向人造血细胞的转移。据报道，在早期临床阶段，粘多糖糖化患者的骨髓移植已停止神经系统恶化的进展。作为第一步，我们产生了重组逆转录病毒，该逆转录病毒表达人β-葡萄糖醛酸酶基因，并将该病毒感染到从脐带血液中纯化的CD34+细胞。使用离心促进感染方案，我们在菌落形成单位粒细胞/巨噬细胞和爆发形成单位红细胞的转导效率（AVE.67％）中得到了转导效率（AVE.67％）。在严重的免疫缺陷小鼠中转导的CD34+细胞是否迁移到大脑中，实验正在进行。