CNS WHITE MATTER TRACTS AS A NOVEL AVENUE FOR GENE THERAPY FOR KRABBE DISEASE

中枢神经系统白质束作为克拉伯病基因治疗的新途径

• 批准号: 8358155
• 负责人: Bruce A. Bunnell
• 金额: $ 2.9万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358155
• 关键词: Affect; Animal Model; Animals; Applications Grants; Behavioral; Brain; Clinical Trials; Data; Diffusion; Disease; Enzymes; Funding; Future; Gene Delivery; Gene Transfer; Genes; Globoid cell leukodystrophy; Grant; Human; Immune response; Injection of therapeutic agent; Internal Capsule; Lentivirus Vector; Macaca mulatta; Microglia; Modeling; Mus; National Center for Research Resources; Neonatal; Outcome; Pathology; Physiological; Preparation; Primates; Principal Investigator; Production; Publishing; Research; Research Infrastructure; Resources; Site; Source; Spinal Cord; Technology; Testing; Tissues; Transgenes; United States National Institutes of Health; cost; design; galactosylceramidase; gene therapy; improved; lentiviral-mediated; nonhuman primate; novel; white matter

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Gene therapy (GT) represents a promising approach for the treatment of the CNS pathology in Lysosomal Storage Disorders (LSD), as it has the potential to provide a permanent source of the deficient enzyme. Our group has been developing a lentiviral (LV)-mediated intracerebral GT to treat Globoid Cell Leukodystrophy (GLD) that could achieve maximal transgene dispersal in the CNS from a limited number of injection sites. Published data from our group (in Preliminary Data section in this proposal) clearly demonstrate that a single lentiviral vector (LV) injection into a highly interconnected white matter region (external/internal capsule; EC/IC) resulted in rapid and robust expression of functional galactocerebrosidase (GALC) throughout the entire brain and spinal cord in the Twitcher mouse (murine model of Krabbe disease), resulting in global rescue of enzymatic activity, and marked decrease of activated microglia. The stable production and widespread distribution of the GALC enzyme achieved following EC/IC injection in affected neonatal mice suggests that this approach effectively produces physiological levels of the missing enzyme. The logical scientific progression in the maturation of this gene delivery technology is to assess this strategy in large animals models, including nonhuman primates (NHPs). Hypothesis: Administration of lentiviral vector expressing GALC to the EC/IC region of the Krabbe-affected primate brain will result in transduction and robust expression throughout the CNS. We will test this hypothesis through two Specific Aims. #1. Determine if LV-mediated delivery of the GALC gene in to the EC/IC improves the behavioral and neuromotor deficits of Krabbe-affected rhesus macaques. #2. To assess gene transfer efficiency, diffusion, distribution and long-term expression of the GALC enzyme in CNS tissues of injected NHP (Krabbe-affected and normal animals), histopathologic outcomes and immune response associated with the white matter-directed gene delivery strategy. As this research moves toward human clinical trials, the proposed research plan is absolutely essential for testing this novel GT approach in a large animal model recapitulating the disease. The data generated from this pilot proposal will be integral for the preparation of future grant proposals and be of significant value in designing human clinical trials.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 基因治疗（GT）代表了溶酶体储存障碍中CNS病理学治疗（LSD）的一种有希望的方法，因为它有可能提供不足酶的永久来源。我们的小组一直在开发慢病毒（LV）介导的脑内GT，以治疗球形细胞白细胞营养不良（GLD），可以从有限数量的注射部位中获得中枢神经系统中的最大转基因分散。我们小组已发布的数据（在本提案的初步数据部分中）清楚地表明，单个慢病毒矢量（LV）注射到高度相互连接的白质区域（外部/内部胶囊； EC/IC）导致功能快速且强大的表达在Twitcher小鼠（Krabbe疾病的鼠模型）中，整个大脑和脊髓中的半乳糖脑溴生酶（GALC），导致全球营救酶活性，并明显减少活化的小胶质细胞。在受影响的新生小鼠中EC/IC注射后达到的GALC酶的稳定产生和广泛的分布表明，这种方法有效地产生了缺失酶的生理水平。该基因输送技术成熟的逻辑科学进步是在包括非人类灵长类动物（NHP）在内的大型动物模型中评估该策略。假设：施用表达galc的慢病毒载体向受Krabbe影响的灵长类动物大脑的EC/IC区域的施用将导致整个中枢神经系统的转导和稳健的表达。 我们将通过两个具体目标检验这一假设。 ＃1。确定LV介导的GALC基因向EC/IC的递送是否可以改善受Krabbe影响的恒河猕猴的行为和神经运动缺陷。 ＃2。为了评估GALC酶在注射NHP（受Krabbe影响和正常动物）的中枢神经系统组织中的GALC酶的扩散，分布和长期表达，组织病理学结局以及与白质导向的基因递送策略相关的免疫反应。 随着这项研究朝着人类的临床试验迈进，提出的研究计划对于在概括该疾病的大型动物模型中测试这种新型GT方法绝对至关重要。该试点建议产生的数据将是准备未来赠​​款建议的组成部分，并且在设计人类临床试验中具有重要价值。