Translational studies on cerebrospinal fluid (CSF)-directed gene therapy for global neurometabolic brain disease

脑脊液（CSF）定向基因治疗全球神经代谢性脑疾病的转化研究

• 批准号: 9763064
• 负责人: JOHN H WOLFE
• 金额: $ 70.37万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9763064
• 关键词: Address; Adverse event; Affect; Alpha-mannosidase; Animal Diseases; Animal Model; Animals; Autopsy; Bathing; Brain; Brain Diseases; Brain imaging; Cell Transplantation; Cell Transplants; Cells; Cerebral Dominance; Cerebral Ventricles; Cerebrospinal Fluid; Clinical; Clinical Trials; Complementary DNA; Defect; Development; Diffusion Magnetic Resonance Imaging; Disease; Disease Marker; Disease Progression; Disease model; Dose; Effectiveness; Endosomes; Enzymes; Extracellular Space; Family Felidae; Felis catus; Gene Delivery; Gene Proteins; Gene Structure; Gene Transduction Agent; Gene therapy trial; Genes; Genetic; Hereditary Disease; Histologic; Human; Imaging Techniques; Immune response; Infusion procedures; Injections; Investigational Therapies; Laboratory Study; Lesion; Liquid substance; Longevity; Lysosomal Storage Diseases; Lysosomes; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Mannose; Measurement; Mediating; Mendelian disorder; Metabolic; Methods; Modeling; Monitor; Mutation; Neuraxis; Neurologic; Oligosaccharides; Organ Transplantation; Pathology; Patients; Protocols documentation; Recovery; Resolution; Rodent; Route; Serum; Severity of illness; Structural Genes; Structure; Testing; Translating; Translations; Vertebral column; adeno-associated viral vector; advanced disease; alpha-Mannosidosis; base; brain cell; brain size; cellular transduction; cisterna magna; comparative; effective therapy; enzyme replacement therapy; enzyme structure; gene correction; gene product; gene therapy; gray matter; human disease; human model; improved; lateral ventricle; mutant; nervous system disorder; non-invasive imaging; receptor mediated endocytosis; response; translational model; translational study; treatment response; treatment strategy; vector; white matter

ABSTRACT A major barrier to effective treatment of the central nervous system (CNS) in most inherited diseases is that pathology is present throughout the brain because the metabolic defect is present in most brain cells. Thus, global distribution of the gene or gene product is required. Several gene therapy strategies are being investigated for treatment of global brain lesions. However, all of the approaches have significant shortcomings, which become apparent in large animal models of human diseases. Development of more effective treatments in these models will facilitate translation into clinical usage. This project will investigate AAV vector mediated gene delivery into the brain by infusion into the cerebrospinal fluid (CSF), which can result in disseminated delivery of a gene in a large animal brain. The disease model to be evaluated is alpha-mannosidosis (AMD) in the cat, a lysosomal storage disorder (LSD) caused by mutations in the lysosomal enzyme (LE) structural gene, alpha-mannosidase (MANB). The strategy for treatment is based on cross-correction, in which transfer of a normal copy of the MANB cDNA into AMD cells results in the metabolic correction of those gene-transduced cells. Furthermore, the genetically corrected cells release normal MANB enzyme, which is taken up by surrounding cells and corrects them metabolically as well. This well-established cross-correction mechanism is the basis for treatments of most LSDs. The disease progression and improvement from treatment will be monitored in living animals by clinical neurological assessment, life-span increases, serum and CSF analyses, and non-invasive brain imaging by magnetic resonance spectroscopy (MRS) and diffusion tensor imaging (DTI). In post-mortem animals, the disease correction will be analyzed quantitatively for distribution of vector transduction, increases in MANB enzymatic activity, reduction of mannose-containing oligosaccharide substrate accumulation, histopathological changes, and potential adverse host responses. We will address important issues affecting translation by investigating strategies to attain more complete correction of the brain: in Aim 1 we will investigate the effects of alternative routes of CSF delivery; in Aim 2 we will investigate the effects of dose escalation on the extent of resolution of pathology; and in Aim 3 we will determine the effectiveness of therapy when initiated at progressively more severe stages of disease to evaluate the potential to ameliorate advanced disease.

抽象的 在大多数遗传疾病中，有效治疗中枢神经系统（CNS）有效治疗的主要障碍是 这种病理存在于整个大脑中，因为大多数脑细胞中存在代谢缺陷。 因此，需要基因或基因产物的全球分布。正在有几种基因治疗策略 研究了全球脑病变的治疗。但是，所有方法都有重要的 缺点，这在大型人类疾病模型中变得显而易见。更多的发展 这些模型中的有效治疗方法将促进转化为临床用法。 该项目将通过输注向大脑进行AAV载体介导的基因输送到大脑 脑脊液（CSF），这可能导致大脑中的基因传播。这 要评估的疾病模型是CAT中的α-甘露糖苷病（AMD），一种溶酶体存储障碍（LSD） 是由溶酶体酶（LE）结构基因，α-甘露糖苷酶（MANB）突变引起的。策略 用于治疗是基于交叉校正的，其中将MANB cDNA的正常副本转移到AMD中 细胞会导致对这些基因转导细胞的代谢校正。此外，遗传纠正 细胞释放正常的MANB酶，该酶被周围细胞占用并将其代谢纠正 出色地。这种完善的跨校正机制是大多数LSD治疗的基础。 疾病的进展和治疗改善将通过临床监测活动物 神经系统评估，寿命增加，血清和CSF分析以及非侵入性大脑成像 磁共振光谱（MRS）和扩散张量成像（DTI）。在验尸动物中， 疾病纠正将进行定量分析以分布载体转导的分布，MANB的增加 酶活性，含甘露糖的寡糖底物积累的减少，组织病理学 变化和潜在的不良宿主反应。 我们将通过调查策略来解决影响翻译的重要问题 对大脑的完全校正：在AIM 1中，我们将研究CSF递送替代途径的影响； 在AIM 2中，我们将研究剂量升级对病理解决程度的影响；在目标3中 我们将在疾病逐渐更严重的疾病阶段启动时确定治疗的有效性 评估改善晚期疾病的潜力。