Gene Transfer and NMR Studies in Alpha-Mannosidosis Brain

α-甘露糖苷沉积症脑中的基因转移和核磁共振研究

• 批准号: 9271721
• 负责人: JOHN H WOLFE
• 金额: $ 60.25万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-03 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9271721
• 关键词: 1 year old; Address; Alpha-mannosidase; Animals; Biological Assay; Biological Markers; Birth; Brain; Brain Diseases; Brain Pathology; Cell Transplants; Cells; Central Nervous System Diseases; Cerebral cortex; Clinical; Clinical Protocols; Clinical Trials; Complementary DNA; Data; Defect; Dependovirus; Diffusion Magnetic Resonance Imaging; Disease; Disease Progression; Disease model; Dose; Endosomes; Ensure; Enzymes; Felis catus; Gene Proteins; Gene Transfer; Genes; Genetic; Histopathology; Human; Image; Imaging Techniques; Inborn Genetic Diseases; Infusion procedures; Injection of therapeutic agent; Investigational Therapies; Life; Longevity; Lysosomal Storage Diseases; Lysosomes; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Mannose; Measures; Mediating; Metabolic; Methods; Modeling; Monitor; Monkeys; Mus; Mutation; Neuraxis; Neurologic; Neurons; Oligosaccharides; Organ; Organ Transplantation; Pathology; Patients; Pattern; Property; Protocols documentation; Recombinant adeno-associated virus (rAAV); Resolution; Rodent; Route; Serotyping; Serum; Specificity; Structural Genes; Structure; Techniques; Therapeutic; Therapeutic Effect; Time; Tissues; Translating; Translations; Treatment Efficacy; Work; adeno-associated viral vector; alpha-Mannosidosis; base; cell type; cellular transduction; gene correction; gene therapy; gene transfer vector; improved; in vivo; mutant; nervous system disorder; non-invasive imaging; novel; receptor; receptor mediated endocytosis; research study; response; scale up; symptom treatment; treatment response; treatment strategy; uptake; vector; vector genome

Gene Transfer and NMR Studies in Alpha-Mannosidosis Brain. We will investigate the ability of systemic delivery of a novel adeno-associated virus (AAV) gene transfer vector to treat the central nervous system (CNS) disease in a cat model of the human lysosomal storage disease (LSD) alpha-mannosidosis (AMD). In most LSDs genetic correction of a relatively small number of mutant cells results in secretion of the normal enzyme and receptor-mediated uptake by surrounding cells, resulting in metabolic correction of the non- transduced cells. The major problem for treating the brain is that pathology is present throughout the CNS because the metabolic defect is present in all cells. Thus, treating the whole brain requires global distribution of the therapeutic normal alpha-mannosidase (MANB) enzyme. This problem is exacerbated by the enormous size difference between mouse and human brains (~3,000 fold). The domestic cat brain is an excellent intermediate in size as it is ~100 times larger than a mouse brain, while the human brain between birth and 1 year (when treatment is expected to be the most effective) is only 10-30 times larger than the cat brain. The cat brain also has a gyrencephalic cerebral cortex that is structurally much more similar to the human brain than the rodent's. Thus, the underlying premise of this translational project is that strategies developed to globally correct the AMD cat brain are more likely to translate effectively into clinical trials. We have shown in this project that AAV gene transfer into the AMD cat brain either by multiple intraparenchymal injections or by infusion into the CSF can improve clinical and histological parameters, but the correction is incomplete. Certain AAVs can enter the CNS after systemic intravascular delivery in mice and mediate widespread transduction, but in large animal brains vector distribution is much more limited. We have developed a novel AAV vector for systemic delivery that transduces neurons throughout the cerebral cortex and other regions of both cat and monkey brains. The new vector also has the novel property of efficiently delivering the gene into large brains with a single-stranded AAV vector genome, whereas previous large animal intravascular AAV delivery experiments have used self-complementary (sc)-AAVs, which are too small to accommodate the MANB cDNA (~3kb). The novel vector will be compared to other AAVs to determine if complimentary patterns of transduction occur in the CNS and other organs to improve the therapeutic effect. We also have developed MR-based imaging assays to non-invasively measure brain pathology and preliminary experiments show they can quantitatively monitor the response to AAV treatment. The specific aims are directed towards optimizing delivery, minimizing dose, evaluating clinical and lifespan improvements, and assessing the accuracy of non- invasive imaging parameters by correlation with histopathology. Although significant progress has been made on this project to date, the treatment is still incomplete and further improvements are needed to ensure the most effective translation into a clinical protocol for human patients.

α-甘露糖苷沉积症脑中的基因转移和核磁共振研究。我们将考察系统性的能力 提供新型腺相关病毒（AAV）基因转移载体来治疗中枢神经系统 人类溶酶体贮积症（LSD）α-甘露糖苷贮积症（AMD）猫模型中的中枢神经系统（CNS）疾病。在 大多数LSD对相对少量的突变细胞进行基因校正，导致正常细胞的分泌 酶和受体介导的周围细胞的摄取，导致非代谢校正 转导的细胞。治疗大脑的主要问题是整个中枢神经系统都存在病变 因为代谢缺陷存在于所有细胞中。因此，治疗整个大脑需要全球分布 治疗性正常α-甘露糖苷酶（MANB）的酶。这个问题因巨大的 小鼠和人类大脑的大小差异（约 3,000 倍）。家养的猫脑子是极好的 中等大小，因为它比小鼠大脑大 100 倍，而人类大脑在出生到 1 岁之间 年（预计治疗最有效时）仅比猫脑大10-30倍。这 猫的大脑也有一个环脑皮质，其结构与人类大脑更加相似 比啮齿动物的。因此，这个转化项目的基本前提是制定战略 全面纠正 AMD 猫脑更有可能有效转化为临床试验。我们已经展示在 该项目通过多次脑实质内注射或通过注射将 AAV 基因转移到 AMD 猫脑中 脑脊液输注可以改善临床和组织学参数，但纠正不完全。 某些 AAV 在小鼠全身血管内递送后可以进入中枢神经系统并介导广泛的 转导，但在大型动物大脑中，载体分布更加有限。我们开发了一本小说 用于全身递送的 AAV 载体，可在整个大脑皮层和大脑其他区域转导神经元 猫脑和猴脑。新载体还具有将基因有效递送到体内的新颖特性。 具有单链 AAV 载体基因组的大型大脑，而之前的大型动物血管内 AAV 递送实验使用了自我互补（sc）-AAV，其太小而无法容纳 MANB cDNA (~3kb)。该新型载体将与其他 AAV 进行比较，以确定是否存在互补模式 中枢神经系统和其他器官中发生转导以提高治疗效果。我们还开发了 基于 MR 的成像分析可无创测量大脑病理学，初步实验表明它们 可以定量监测 AAV 治疗的反应。具体目标是优化 交付，最大限度地减少剂量，评估临床和寿命的改善，并评估非-的准确性 通过与组织病理学相关的侵入性成像参数。尽管已经取得了重大进展 迄今为止，该项目的处理仍然不彻底，需要进一步改进以确保 最有效地转化为人类患者的临床方案。