Delivery of Therapeutic Enzymes and Genes to the CNS

将治疗酶和基因递送至中枢神经系统

• 批准号: 6615852
• 负责人: JAKOB REISER
• 金额: $ 23.61万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6615852
• 关键词: Krabbe's disease; Lentivirus; behavior test; biotherapeutic agent; blood brain barrier; central nervous system; enzyme activity; gene delivery system; gene expression; gene therapy; genetically modified animals; glucosylceramidase; histopathology; human immunodeficiency virus 1; immunocytochemistry; laboratory mouse; laboratory rat; magnetic resonance imaging; peripheral nervous system; polymerase chain reaction; positron emission tomography; transfection /expression vector; western blottings

DESCRIPTION (provided by applicant): Owing to the structural and functional complexity and relative inaccessibility of the central nervous system (CNS), most chronic neurologic and neurodegenerative disorders lack effective treatments, and, therefore, require the development of novel therapeutic approaches including gene therapy and cellular therapy. Krabbe disease, or globoid cell leukodystrophy (GLD), is an inherited, recessive disorder affecting the peripheral nervous system (PNS) and the CNS caused by defects of the lysosomal enzyme galactocerebrosidase (GALC). This enzyme catalyzes the lysosomal hydrolysis of galactosylceramide and galactosylsphingosine (psychosine). The enzyme defect causes a series of pathological changes including demyelination. This autosomal recessive disease affects humans and animals including dogs, mice, and rhesus monkeys. The goal of the proposed project is to explore gene therapy strategies for Krabbe disease using improved lentivirus-based gene and protein delivery strategies. These strategies will initially be tested in rats and later on extended to mice and rhesus monkeys affected with GLD. The Specific Aims are: 1. To modify HIV-l-based lentiviral vectors for improved transgene delivery and expression in the CNS. These vectors will harbor constitutive as well as regulatable promoters. Vectors will be pseudotyped with the vesicular stomatitis virus (VSV)-G glycoprotein. Pseudotypes involving the Mokola virus G glycoprotein will also be tested with a view toward distributing such vectors more globally in the CNS. Vector delivery will be carried out by direct brain injection. 2. To investigate protein transduction mechanisms with a view toward facilitating the global delivery in the CNS of GALC tagged with the transduction domain of HIV-1 TAT, the membrane translocating hydrophobic sequence from fibroblast growth factor or the herpes simplex virus VP22 protein. 3. To use lentiviral vectors to transfer the GALC cDNA into the brains of mice and rhesus monkeys affected with GLD to evaluate the capacity of such vectors to correct the GLD defect and histological abnormalities.

描述（由申请人提供）： 由于中枢神经系统（CNS）的结构和功能复杂性以及相对无法访问，大多数慢性神经系统和神经退行性疾病缺乏有效的治疗，因此需要开发新的治疗方法，包括基因疗法和细胞治疗。 Krabbe病或Globoid细胞白细胞营养不良（GLD）是一种影响周围神经系统（PNS）的遗传性隐性疾病，以及由溶酶体半乳糖核苷酶（GALC）缺陷引起的CNS。这酶催化了半乳糖酰胺和半乳糖基肾上腺素（Psychosine）的溶酶体水解。酶缺陷会导致一系列病理变化，包括脱髓鞘。这种常染色体隐性疾病会影响人类和动物，包括狗，小鼠和恒河猴。拟议项目的目的是使用改进的基于慢病毒的基因和蛋白质递送策略来探索Krabbe疾病的基因治疗策略。这些策略最初将在大鼠中进行测试，然后在扩展到受GLD影响的小鼠和恒河猴上进行测试。具体目的是：1。修改基于HIV-L的慢病毒载体，以改善CNS中的转基因递送和表达。这些向量将具有构成和可调节的启动子。载体将用囊泡口腔炎病毒（VSV）-G糖蛋白进行伪型。涉及Mokola病毒G糖蛋白的假型也将进行测试，以期在CNS中在全球范围内更加分发此类向量。矢量输送将通过直接脑注射进行。 2。为了研究蛋白质转导机制，以促进用HIV-1 TAT的转导域标记的GALC中的全局传递，膜从成纤维细胞生长因子或Herpes Simplex Simplex病毒VP22蛋白质转移了疏水性序列。 3。使用慢病毒载体将galc cDNA转移到受GLD影响的小鼠和恒河猴的大脑中，以评估此类向量纠正GLD缺陷和组织学异常的能力。