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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

基本信息

批准号：
7030557
负责人：
HAIYAN FU
金额：
$ 17.04万
依托单位：
RESEARCH INST NATIONWIDE CHILDREN'S HOSP
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-08-15 至 2006-06-30
项目状态：
已结题

项目摘要

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）：粘多糖沉积症III B（Sanfilippo 13型，MPS IIIB）是一种由于α-N-乙酰氨基葡萄糖苷酶（NaGlu）遗传性缺乏引起的溶酶体贮积症。该病的特征是轻度躯体疾病，大多数患者在6-10岁时出现严重的神经变性，社会和适应能力迅速和进行性恶化，导致过早死亡。MPS III B患者的中枢神经系统（CNS）疾病目前尚无治疗方法，这通常是过早死亡的原因。腺相关病毒（Adeno-associated virus，AAV）是一种具有广泛感染性的基因传递系统，其致病机制尚不清楚。在本项目中，将使用基因敲除小鼠模型研究AAV介导的MPS III B CNS疾病的基因治疗。已经构建了两种AAV载体，其含有由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，并且已经显示出功能性NaGlu的有效表达以及通过重组NaGlu在体外MPS IIIB细胞培养物中和体内小鼠脑中对溶酶体储存的校正。通过直接显微注射将载体递送至MPS III B小鼠的多个脑区，以研究rNaGlu的更有效体内表达和分布以及注射后脑中溶酶体蓄积的校正。此外，将进行研究以开发更有效的基因递送到脑中的方法，因为基因治疗载体在脑中的有限分布是CNS治疗的最大障碍之一。通过直接显微注射将含有增强的绿色荧光蛋白基因的AAV载体递送到多个小鼠脑区域来实现，以a）比较由不同AAV血清型介导的基因表达的分布（1、2和5）载体; B）研究通过不同注射途径，特别是外周递送递送到小鼠脑中的基因表达的分散;和c）通过修饰AAV病毒载体的递送介质来研究AAV载体的传播; d）.通过单次注射进行多部位输注，以增加单次注射后AAV载体的分布。本项目的目的是研究使用AAV介导的基因治疗治疗MPS III B儿童CNS疾病的可行性，并使用小鼠模型实现AAV载体在CNS中的广泛分布。本项目的长期目标是开发治疗MPS IIIB患者CNS疾病的新疗法。