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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：
7785865
负责人：
MIGUEL S ESTEVES
金额：
$ 52.71万
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/7785865
关键词：
2 year old Acids 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 Galactosidase 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 Research Personnel Serotyping Severity of illness Single-Gene Defect Spinal Cord Staging Surface Testing Thalamic structure Therapeutic Therapeutic Studies Time Treatment Efficacy Tropism Tyrosine Validation adeno-associated viral vector base 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 ¿-galactosidase (¿gal), 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 ¿-galactosidase cDNA, although AAV vectors for human clinical trials will express human ¿gal. Therefore, it is necessary to perform bio-equivalency studies to ensure the functionality of the human ¿gal 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 ¿gal 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. PROJECT NARRATIVE: The AAV vector-based approaches investigated in this project for CNS therapy in GM1-gangliosidosis are likely to be applicable to many, if not all neurodegenerative lysosomal storage diseases. The experiments proposed here are essential to determine the scalability and therapeutic efficacy of each approach applied to a much larger and complex brain such as that in GM1-gangliosidosis cats. Moreover the AAV vectors encoding the human lysosomal ¿-galactosidase enzyme developed and tested here for their therapeutic efficacy in GM1 mice and cats may ultimately progress into human clinical trials for this devastating disease for which there is no treatment. Finally, the investigators are proposing to develop a new generation of AAV vectors capable of targeting the brain through the vasculature. This new generation of AAV vectors may provide the means to develop highly effective gene therapy approaches to treat many childhood, adult, as well as geriatric neurological diseases currently beyond the reach of modern medicine.

描述（由申请人提供）：溶酶体贮积病（LSD）是单基因缺陷，导致分解代谢溶酶体酶的缺乏以及其一种或多种大分子底物在溶酶体中的积累。 GM1 神经节苷脂沉积症是由于溶酶体酶 ¡-半乳糖苷酶 (¡gal) 缺乏所致，导致进行性和致命的神经变性。根据疾病严重程度和发病年龄，人类 GM1 神经节苷脂贮积症分为三种类型，分为婴儿型、青少年型和成人型。基于敲除小鼠模型中非凡的初步结果，使用腺相关病毒载体进行基因治疗是治疗 GM1 神经节苷脂沉积症的一种非常有前途的策略。然而，小鼠大脑比人类婴儿大脑小 1,000-2,000 倍，复杂性也低得多，因此需要在小鼠中获得的结果在大脑大小和复杂性与人类更相似的动物中重现。已充分表征的 GM1 神经节苷脂沉积症猫科动物模型的大脑尺寸仅比人类婴儿小 15 倍，将用于测试 AAV 载体在更大、更复杂的大脑中的治疗效果。此外，初步的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β-半乳糖苷酶。因此，在开始人体临床试验之前，有必要进行生物等效性研究，以确保人gal cDNA的功能性。本申请中的实验旨在通过以下具体目标将 AAV 基因治疗推向人体临床试验。在目标 1 中，将在实质内或脑脊液介导的 AAV 载体递送至猫科动物 GM1 大脑后进行短期治疗研究。此外，还将在 GM1 小鼠的短期实验中对人类和小鼠 gal 进行直接比较。在 Aim 2 中，将利用疾病进展的临床、行为和生化检测来对 GM1 猫和小鼠的治疗效果进行长期研究。最后，Aim 3将测试AAV衣壳库的体内选择原理（分子进化），以鉴定血管内（iv）输注后对GM1小鼠中枢神经系统（CNS）具有趋向性的AAV载体，然后测试它们在这些小鼠中的治疗效果。验证这种在静脉输注后生成具有 CNS 向性的 AAV 载体的方法将为开发此类载体用于人类应用铺平道路。项目叙述：本项目中研究的基于 AAV 载体的 GM1 神经节苷脂沉积症中枢神经系统治疗方法可能适用于许多（如果不是全部）神经退行性溶酶体贮积病。这里提出的实验对于确定应用于更大和复杂的大脑（例如 GM1 神经节苷脂沉积症猫的大脑）的每种方法的可扩展性和治疗效果至关重要。此外，这里开发并测试的编码人类溶酶体β-半乳糖苷酶的AAV载体在GM1小鼠和猫中的治疗效果可能最终进入针对这种无法治疗的破坏性疾病的人体临床试验。最后，研究人员提议开发新一代 AAV 载体，能够通过脉管系统靶向大脑。新一代 AAV 载体可能提供开发高效基因治疗方法的手段，以治疗目前现代医学无法达到的许多儿童、成人以及老年神经系统疾病。