Gene Therapy for Neurodegenerative Lysosomal Storage Diseases

神经退行性溶酶体贮积病的基因治疗

基本信息

批准号：
8017404
负责人：
MIGUEL S ESTEVES
金额：
$ 50.14万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8017404
关键词：
2 year old Adolescent Adult Age of Onset Age-Months Animal Model Animals Behavioral Biochemical Biological Assay Brain Capsid Cells Cerebellar Nuclei Cerebrospinal Fluid Childhood Clinical Clinical Trials Clinical assessments Complementary DNA Complex DNA Shuffling Development Disease Disease Progression Dose Effectiveness Elderly Ensure Enzymes Family Felidae Felis catus Frequencies Functional disorder Future Ganglioside GM1 Gangliosidosis GM1 Gene Delivery Gene Transfer Generations Genes Goals Health Human Infant Infusion procedures Knockout Mice Libraries Live Birth Longitudinal Studies Lung Lysosomal Storage Diseases Lysosomes Magnetic Resonance Imaging Mediating Methodology Modality Modeling Modern Medicine Molecular Evolution Mus Mutation Nerve Degeneration Neuraxis Neurologic Neurons Neurosciences Organ Outcome Patients Peripheral Phase Process Serotyping Severity of illness Single-Gene Defect Spinal Cord Staging Surface Testing Thalamic structure Therapeutic Therapeutic Studies Time Treatment Efficacy Tropism Tyrosine Validation acid beta-galactosidase adeno-associated viral vector base beta-Galactosidase brain size design effective therapy efficacy testing gene therapy in vivo infancy lateral ventricle meetings minimally invasive mouse model mutant nervous system disorder neurochemistry novel strategies research study therapeutic effectiveness transduction efficiency treatment strategy vector

项目摘要

DESCRIPTION (Provided by Applicant): Lysosomal storage diseases (LSDs) are single gene defects that result in the deficiency of a catabolic lysosomal enzyme and accumulation of one or more of its macromolecular substrates in the lysosome. GM1 gangliosidosis results from deficiency of the lysosomal enzyme beta-galactosidase (betagal), leading to progressive and fatal neurodegeneration. Human GM1 gangliosidosis occurs as three types based on disease severity and age of onset, classified as infantile, juvenile, and adult-onset forms. Gene therapy with adeno-associated viral vectors is a very promising strategy for treatment of GM1 gangliosidosis based on extraordinary preliminary results in the knockout mouse model. However, the mouse brain is 1,000-2,000 times smaller and much less complex than the brain of a human infant, requiring that results obtained in mice be reproduced in an animal with a brain size and complexity more similar to humans. The well-characterized feline model of GM1 gangliosidosis, with a brain size only 15 times smaller than a human infant, will be utilized to test therapeutic efficacy of AAV vectors in a larger and more complex brain. In addition, preliminary mouse studies employed a mouse beta-galactosidase cDNA, although AAV vectors for human clinical trials will express human betagal. Therefore, it is necessary to perform bio-equivalency studies to ensure the functionality of the human betagal cDNA before initiating human clinical trials. The experiments in this application are designed to advance AAV gene therapy toward human clinical trials through the following specific aims. In Aim 1, short-term therapeutic studies will be performed after intraparenchymal or CSF-mediated delivery of AAV vectors to the feline GM1 brain. Also, a direct comparison of human and mouse betagal will be conducted in short-term experiments in GM1 mice. In Aim 2 long-term studies of therapeutic efficacy in GM1 cats and mice will be conducted using clinical, behavioral, and biochemical assays of disease progression. Finally, Aim 3 will test the principle of in vivo selection of an AAV capsid library (molecular evolution) to identify AAV vectors with tropism for the GM1 mouse central nervous system (CNS) after intravascular (iv) infusion, and then test their therapeutic efficacy in these mice. Validation of this approach to generate AAV vectors with CNS-tropism after iv infusion will pave the way for development of such vectors for human application.

描述（由申请人提供）：溶酶体储存疾病（LSD）是单个基因缺陷，导致分解代谢溶酶体酶的缺乏，并积累了其一种或多种溶酶体中其大分子分子底物。 GM1神经节病是由溶酶体β-半乳糖苷酶（β）的缺乏引起的，导致进行性和致命的神经变性。人GM1神经节病以三种类型的形式出现，基于疾病的严重程度和发病年龄，被归类为婴儿，少年和成人发作形式。与腺相关病毒载体的基因治疗是基于敲除小鼠模型中特殊初步结果治疗GM1神经节病的非常有前途的策略。但是，小鼠大脑比人类婴儿的大脑小1,000-2,000倍，并且需要在小鼠中获得的结果在大脑大小和复杂性的动物中得到复制。 GM1神经节病的特征良好的猫科动物模型，其大脑大小仅比人类婴儿小15倍，可用于测试AAV载体在更大，更复杂的大脑中的治疗功效。此外，初步小鼠研究采用了小鼠β-半乳糖苷酶cDNA，尽管用于人类临床试验的AAV载体将表达人类betagal。因此，有必要进行生物等效性研究，以确保人类betagal cDNA在启动人类临床试验之前的功能。本应用中的实验旨在通过以下特定目的将AAV基因治疗推向人类临床试验。在AIM 1中，将在掌内部或CSF介导的AAV向量向猫GM1脑的递送后进行短期治疗研究。此外，在GM1小鼠的短期实验中将进行人类和小鼠β的直接比较。在AIM 2中，将使用疾病进展的临床，行为和生化测定法对GM1猫和小鼠的治疗功效进行长期研究。最后，AIM 3将测试AAV Capsid库（分子进化）的体内选择的原理，以鉴定血管内（IV）输注后GM1小鼠中枢神经系统（CNS）的AAV向量，然后测试其在这些小鼠中的治疗功效。验证这种方法以在IV输注后生成CNS-Tropism的AAV向量将为开发这种向量的人类应用铺平道路。