CNS Gene Therapy for Mucopolysaccharidosis IIIB w/ AAV

使用 AAV 治疗粘多糖贮积症 IIIB 的中枢神经系统基因疗法

• 批准号: 6768655
• 负责人: HAIYAN FU
• 金额: $ 6.43万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-15 至 2004-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6768655
• 关键词: N acetylglucosaminidase; adeno associated virus group; behavior test; cell line; central nervous system; cerebral ventricles; complementary DNA; enzyme activity; fluorescence microscopy; gene expression; gene targeting; gene therapy; genetically modified animals; histochemistry /cytochemistry; inborn lysosomal enzyme disorder; laboratory mouse; mucopolysaccharidosis type III; nonhuman therapy evaluation; thalamus; transfection; transfection /expression vector; transmission electron microscopy

DESCRIPTION (provided by applicant): Mucopolysaccharidosis III B (Sanfilippo type 13, MPS IIIB) is a lysosomal storage disorder due to the inherited deficiency of a-N-acetylglucosaminidase (NaGlu). The disease is characterized by mild somatic disease with severe neurological degeneration in most of patients by 6-10 years of age with rapid and progressive deterioration of social and adaptive abilities leading to premature death. No treatment is currently available for the central nervous system (CNS) disorder of MPS III B patients, which is usually the cause of premature death. Adeno-associated virus (AAV) has been shown to provide a promising gene delivery system for its ability of infecting wide range of tissues/organs and with no known pathogenesis in human. In this project, AAV-mediated gene therapy for the CNS disease of MPS III B is to be studied using a knock-out mouse model. Two AAV-vectors, containing human NaGlu cDNA driven by a CW promoter or the endogenous brain promoter, neuron specific enolase (NSE) promoter, have been constructed and have shown efficient expression of functional NaGlu and the correction of lysosomal storage by recombinant NaGlu in vitro in MPS IIIB cell cultures and in vivo in mouse brain. The vectors are to be delivered into multiple brain areas of MPS III B mice by direct microinjection, to study more efficient in vivo expression and distribution of rNaGlu and the correction of lysosomal storage in the brain after the injection. In addition, studies will be conducted to develop more efficient means of gene delivery into brain because limited distribution of gene therapy vectors in brain is one of the biggest obstacles for CNS therapies. It will be achieved by delivering AAV vectors containing an enhanced green fluorescent protein gene into multiple mouse brain areas by direct microinjection, to a) compare the distribution of gene expression mediated by different AAV serotype (1, 2 and 5) vectors; b) study the dispersion of gene expression delivered into mouse brain through different injection route, especially peripheral delivery; and c) study the spread of AAV vectors by modifying the delivery media of AAV viral vectors; d). Conduct multiple site infusion by a single injection, to increase the distribution of AAV vectors after a single injection. The goal of this project is to study the feasibility of using AAV-mediated gene therapy to treat the CNS disease in MPS III B children and to achieve broad distribution of AAV vectors in CNS, using mouse model. The long-term goal of this project is to develop novel therapy for CNS disease in MPS IIIB patients.

描述（由申请人提供）： 粘多糖贮积症 III B（Sanfilippo 13 型，MPS IIIB）是一种由于 α-N-乙酰氨基葡萄糖苷酶 (NaGlu) 遗传性缺陷所致的溶酶体贮积症。该疾病的特点是，大多数 6-10 岁患者会出现轻度躯体疾病，伴有严重的神经退行性变，社交和适应能力迅速进行性恶化，导致过早死亡。目前尚无针对 MPS III B 患者中枢神经系统 (CNS) 疾病的治疗方法，该疾病通常是导致过早死亡的原因。腺相关病毒（AAV）已被证明提供了一种有前途的基因传递系统，因为它能够感染广泛的组织/器官，并且在人类中的发病机制尚不清楚。在该项目中，将使用基因敲除小鼠模型来研究 AAV 介导的 MPS III B 中枢神经系统疾病的基因治疗。已经构建了两个 AAV 载体，包含由 CW 启动子或内源性脑启动子、神经元特异性烯醇化酶 (NSE) 启动子驱动的人 NaGlu cDNA，并显示出功能性 NaGlu 的有效表达以及重组 NaGlu 在体外校正溶酶体储存MPS IIIB 细胞培养物和小鼠大脑体内。该载体将通过直接显微注射递送至 MPS III B 小鼠的多个大脑区域，以研究 rNaGlu 更有效的体内表达和分布以及注射后大脑中溶酶体储存的校正。此外，还将进行研究以开发更有效的基因递送至大脑的方法，因为基因治疗载体在大脑中的有限分布是中枢神经系统治疗的最大障碍之一。它将通过直接显微注射将含有增强型绿色荧光蛋白基因的 AAV 载体递送到多个小鼠大脑区域来实现，以 a) 比较不同 AAV 血清型（1、2 和 5）载体介导的基因表达分布； b) 研究通过不同注射途径递送至小鼠大脑中的基因表达的分散性，特别是外周递送； c) 通过修改AAV病毒载体的传递介质来研究AAV载体的传播； d).单次注射多部位输注，增加单次注射后AAV载体的分布。该项目的目标是研究使用 AAV 介导的基因疗法治疗 MPS III B 儿童中枢神经系统疾病的可行性，并使用小鼠模型实现 AAV 载体在中枢神经系统中的广泛分布。该项目的长期目标是开发针对 MPS IIIB 患者中枢神经系统疾病的新疗法。