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

基本信息

批准号：
10379947
负责人：
JOHN H WOLFE
金额：
$ 63.92万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
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/10379947
关键词：
Address Adverse event Affect Alpha-mannosidase Animal Diseases Animal Model Animals Autopsy Bathing Brain Brain Diseases Cell Transplantation 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 Endosomes Enzymes Extracellular Space Family Felidae Felis catus Gene Delivery Gene Proteins 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 magnetic resonance imaging brain size cellular transduction cisterna magna comparative effective therapy effectiveness evaluation enzyme replacement therapy 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

项目摘要

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性疾病中，有效治疗中枢神经系统 (CNS) 的一个主要障碍是这种病理现象存在于整个大脑中，因为代谢缺陷存在于大多数脑细胞中。因此，需要基因或基因产物的全球分布。几种基因治疗策略正在研究中研究用于治疗全脑损伤。然而，所有的方法都有显着的意义这些缺点在人类疾病的大型动物模型中变得显而易见。开发更多这些模型中的有效治疗将有助于转化为临床应用。该项目将研究 AAV 载体介导的基因通过输注到大脑中的过程脑脊液 (CSF)，可导致基因在大型动物大脑中传播。这待评估的疾病模型是猫的α-甘露糖苷贮积症（AMD），一种溶酶体贮积症（LSD）由溶酶体酶 (LE) 结构基因 α-甘露糖苷酶 (MANB) 突变引起。策略治疗基于交叉校正，其中将正常的 MANB cDNA 拷贝转移到 AMD 细胞导致这些基因转导细胞的代谢校正。此外，经过基因校正细胞释放正常的 MANB 酶，该酶被周围细胞吸收并以代谢方式纠正它们出色地。这种完善的交叉校正机制是大多数 LSD 治疗的基础。将通过临床监测活体动物的疾病进展和治疗改善神经学评估、寿命延长、血清和脑脊液分析以及非侵入性脑成像磁共振波谱（MRS）和扩散张量成像（DTI）。在动物死后，对疾病校正进行定量分析，了解载体转导的分布、MANB 的增加酶活性，减少含甘露糖寡糖底物积累，组织病理学变化和潜在的不良宿主反应。我们将通过研究策略来解决影响翻译的重要问题，以实现更多目标完全纠正大脑：在目标 1 中，我们将研究脑脊液输送替代途径的影响；在目标 2 中，我们将研究剂量递增对病理消退程度的影响；在目标 3 中我们将在疾病逐渐严重的阶段开始确定治疗的有效性评估改善晚期疾病的潜力。