New Treatment of the Brain in Niemann Pick C

Niemann Pick C 的大脑新疗法

基本信息

批准号：
9331841
负责人：
William M Pardridge
金额：
$ 46.1万
依托单位：
LIPOGENE COMPANY, INC.
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-15 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9331841
关键词：
Adult Age Alpha-galactosidase Ataxia Behavioral Biochemical Biomedical Technology Blood Blood - brain barrier anatomy Body Weight Brain Breeding Cell Culture Techniques Cell Density Cells Cessation of life Cholesterol Chronic Clinical Complementary DNA DNA DNA cassette DNA delivery Data Dementia Deposition Development Disease Disease model Embryo Encapsulated Engineering Enzymes Equus caballus Experimental Models Fibroblasts Filipin Fluorescence Microscopy Gait Galactosidase Gene Delivery Gene Expression Genes Genome Goals Grant Histocytochemistry Histology Homozygote Hour Human Inherited Injection of therapeutic agent Intracellular Membranes Intravenous Knockout Mice Laboratories Lipids Liposomes Liver Lysosomal Storage Diseases Maintenance Malignant neoplasm of brain Measures Mediating Medicine Membrane Transport Proteins Messenger RNA Methods Molecular Biology Monoclonal Antibodies Mus Mutate Mutation National Institute of Neurological Disorders and Stroke Nerve Degeneration Neuraxis Neurons Niemann-Pick Diseases Northern Blotting Nuclear Organ Organ Weight Parkinson Disease Peripheral Phase Proteins Purkinje Cells RNA Rare Diseases Reaction Research Retina Seizures Services Small Business Innovation Research Grant Spleen Symptoms System TFRC gene Technology Testing Therapeutic Therapeutic Effect Toxic effect Transgenes Transgenic Mice Translating Treatment Efficacy Viral Vector Western Blotting Work astrogliosis base beta-Galactosidase brain cell cholesterol transporters disease-causing mutation enzyme replacement therapy gene therapy immunoreaction in vivo liposomal delivery molecular trojan horse mouse model neural model new technology nonhuman primate novel strategies novel therapeutics plasmid DNA receptor receptor mediated endocytosis relating to nervous system technology development therapeutic gene transcytosis transgene expression virtual

项目摘要

Abstract Significance: Niemann-Pick Type C1 disease (NPC1) is a devastating inherited neurodegenerative lysosomal storage disease caused by mutations in the NPC1 gene, which encodes for an intracellular membrane cholesterol transporter. Currently, there are few treatment options for NPC1. Protein replacement therapy is not possible, because the NPC1 protein is an insoluble membrane transporter. Gene therapy of either human NPC1, or a long standing mouse model of NPC1, has not been attempted, owing to the large size of the NPC1 mRNA, which exceeds the capacity of common gene therapy viral vectors. In contrast, large size genes can be encapsulated in nonviral delivery systems such as Trojan horse liposomes (THLs). THLs are manufactured with a receptor-specific monoclonal antibody (MAb), which acts as a molecular Trojan horse to ferry the THL from blood into the nuclear compartment of brain cells. In prior work, THL-mediated delivery of plasmid DNA has produced therapeutic effects in mouse models of neural disease including, Parkinson's disease, brain cancer, or lysosomal storage disease. Hypothesis: The hypothesis tested in the present work is that the NPC1 gene can be effectively replaced in brain, and in peripheral organs, with regular intravenous (IV) administration of THLs encapsulating plasmid DNA encoding the NPC1 gene. This hypothesis is supported by prior work in the NPC1 mouse model wherein embryos were transfected with the wild type NPC1 gene and these transgenic mice were cross-bred with the NPC1 mouse. Replacement of the NPC1 gene in brain effectively cured the disease in the NPC1 mouse. Preliminary Data: Prior work with a lysosomal storage disease mouse model has shown it is possible to achieve replacement of the wild type gene in mouse brain with IV administration of THLs carrying the expression plasmid DNA encoding the lysosomal enzyme gene. The THLs are targeted with a MAb specific for the mouse transferrin receptor (TfR). Prior work has also shown that repeat, chronic IV administration of THLs causes no toxicity or immune reactions. Specific Aims: First, THLs will be manufactured, and these THLs are targeted with the MAb specific for the mouse TfR, and encapsulate an expression plasmid DNA encoding for the NPC1 cDNA. The expression plasmid DNA will be engineered with methods used previously for therapeutic plasmid DNA. Second, the potency of the THLs will be assessed in cell culture using human NPC1 fibroblasts. Third, a colony of NPC1 mice will be generated for this project producing at least 36 homozygote mice. The mice will undergo treatment for 12 weeks with weekly IV administration of THLs carrying the NPC1 DNA, beginning at the age of 6 weeks. Treatment efficacy will be assessed with survival, body and organ weights, brain histology, peripheral organ histology, as well as brain biochemical parameters of NPC1 gene expression. This research can be translated to human NPC1, because prior work as shown it is possible to deliver transgenes to virtually all cells in the brain of the adult non-human primate with IV administration of THLs.

抽象的意义：尼曼-匹克 C1 型疾病 (NPC1) 是一种破坏性遗传性神经退行性溶酶体疾病由 NPC1 基因突变引起的贮藏病，该基因编码细胞内膜胆固醇转运蛋白。目前，NPC1 的治疗方案很少。蛋白质替代疗法是不可能，因为 NPC1 蛋白是一种不溶性膜转运蛋白。任一人类的基因治疗由于 NPC1 体积较大，尚未尝试 NPC1 或 NPC1 的长期小鼠模型 mRNA，超出了常见基因治疗病毒载体的容量。相反，大尺寸基因可以封装在非病毒传递系统中，例如特洛伊木马脂质体（THL）。 THL 已制造与受体特异性单克隆抗体 (MAb) 结合，充当分子特洛伊木马来运送 THL 从血液进入脑细胞的核室。在之前的工作中，THL 介导的质粒 DNA 递送在小鼠神经疾病模型中产生了治疗效果，包括帕金森病、脑病癌症或溶酶体贮积病。假设：本工作中检验的假设是 NPC1 通过定期静脉（IV）给药，可以有效地替换大脑和外周器官中的基因 THL 封装编码 NPC1 基因的质粒 DNA。这一假设得到了先前工作的支持 NPC1小鼠模型，其中胚胎转染了野生型NPC1基因，并且这些转基因小鼠与 NPC1 小鼠杂交。大脑中NPC1基因的替换有效治愈了 NPC1 小鼠的疾病。初步数据：之前对溶酶体贮积病小鼠模型的研究表明表明通过静脉注射 THL 可以实现小鼠大脑中野生型基因的替换携带编码溶酶体酶基因的表达质粒DNA。 THL 的目标是对小鼠转铁蛋白受体 (TfR) 具有特异性的 MAb。先前的工作还表明，重复的、慢性的静脉注射 THL 的给药不会引起毒性或免疫反应。具体目标：首先，THL 将这些 THL 是针对小鼠 TfR 特异的 MAb 进行靶向的，并封装了编码NPC1 cDNA的表达质粒DNA。表达质粒DNA将被设计为以前用于治疗性质粒 DNA 的方法。其次，THL 的效力将在使用人 NPC1 成纤维细胞进行细胞培养。第三，该项目将生成一群 NPC1 小鼠产生至少 36 只纯合子小鼠。小鼠将接受为期 12 周的治疗，每周静脉注射从 6 周龄开始施用携带 NPC1 DNA 的 THL。治疗效果将是通过存活率、体重和器官重量、脑组织学、周围器官组织学以及脑组织学进行评估 NPC1基因表达的生化参数。这项研究可以转化为人类 NPC1，因为先前的工作表明，可以将转基因传递到成年非人类大脑中的几乎所有细胞灵长类动物静脉注射 THL。