GENE TRANSFER TO AND EXPRESSION IN NEURONS IN VIVO

体内神经元的基因转移和表达

• 批准号: 3416185
• 负责人: NIGEL William FRASER
• 金额: $ 19.19万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-04-07 至 1994-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3416185
• 关键词: O glycosidase; disease /disorder model; gene expression; gene therapy; genetic manipulation; genetic regulation; herpes simplex virus 1; histochemistry /cytochemistry; in situ hybridization; inborn lysosomal enzyme disorder; laboratory mouse; mucopolysaccharidosis; neurons; recombinant DNA

Developmental neurogenetic diseases can only be studied, and potentially treated, once systems to deliver and express genes in neuronal cells are developed. We will utilize herpes simplex virus (HSV) as a vector for expressing genes in neuronal cells. This human DNA virus, for which about 90% of the U.S. population is seropositive, forms latent infections which persist for the lifetime of the individual. Using HSV derived vectors, we will study the nervous system component of a lysosomal storage disease. Lysosomal storage in neurons results in severe mental retardation in most children with these diseases. Specifically, we will use a bets- glucuronidase-negative-mouse (gusmps/gusmps), with mucopolysacharidosis (MPS) type VII (Sly disease), as an animal model. Experiments will be performed to insert a beta-glucuronidase gene into HSV- 1, under control of the viral promoter that is active during latency. This is the only HSV-1 promoter known to be active long-term in vivo. Animals will be infected with recombinant virus, or with a HSV-1 derived amplicon vector, and beta-glucuronidase gene expression will be studied. The ability of beta-glucuronidase enzyme expressed in latently infected animals to alter lysosomal storage during the life of these animals will be monitored histologically and biochemically. Both the HSV vector and the beta-glucuronidase gene will be engineered for maximal expression and spread of enzyme activity throughout the nervous system. The goal of these studies is to address the problem of treatment of neurogenetic diseases by developing a method of expressing genes in the nervous system in vivo using a ubiquitous human virus. These studies will also provide a method for studying gene regulation in neuronal cells in vivo using the state of the art techniques of molecular biology.

只能研究发育性神经遗传疾病，并有可能 经过处理，一旦在神经元细胞中传递和表达基因的系统是 发达。 我们将利用单纯疱疹病毒（HSV）作为矢量 在神经元细胞中表达基因。 这种人类DNA病毒，大约 90％的美国人群是血清阳性的，形成了潜在感染 坚持个人的一生。 使用HSV派生向量，我们 将研究溶酶体储存疾病的神经系统成分。 在大多数 这些疾病的孩子。 具体来说，我们将使用一个赌注 - 葡萄糖醛酸酶阴性鼠（GUSMPS/GUSMPS），粘膜可刺治疗 （MPS）VII型（狡猾的疾病），作为动物模型。 将进行实验以将β-葡萄糖醛酸酶基因插入HSV- 1，在潜伏期活跃的病毒启动子的控制下。 这 是唯一已知在体内长期活跃的HSV-1启动子。 动物 将被重组病毒或HSV-1衍生的扩增子感染 将研究载体和β-葡萄糖醛酸酶基因表达。 这 在潜在感染动物中表达的β-葡萄糖醛酸苷酶的能力 在这些动物的生命中改变溶酶体储存将是 通过组织学和生化监测。 HSV向量和 β-葡萄糖醛酸酶基因将用于最大表达和 酶活性在整个神经系统中的传播。 这些研究的目的是解决治疗的问题 神经遗传疾病通过开发一种表达基因的方法 神经系统在体内使用无处不在的人类病毒。 这些研究会 还提供了一种研究神经元细胞中基因调节的方法 使用分子生物学的最先进技术的体内。