Gene therapy with modified GlcNAc-1-phosphotransferase for mucolipidosis

使用修饰的 GlcNAc-1-磷酸转移酶基因治疗粘脂沉积症

基本信息

批准号：
10317695
负责人：
PATRICIA I DICKSON
金额：
$ 43.31万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10317695
关键词：
Address Age Amino Acids Behavior Binding Sites Biochemical Biodistribution Biological Assay Blood Circulation Brain Cessation of life Cleaved cell DNA Modification Methylases Data Disease Enzymes Excision Failure Female Genetic Diseases Glucosamine Golgi Apparatus Growth Human Hydrolase I-Cell Disease Impairment In Vitro Inherited Intravenous Knockout Mice Laboratories Lead Lipomucopolysaccharidoses Lysosomes Mannose Mus N-acetylglucopyranosylamine Organ Pathogenicity Pathologic Pathology Patients Pattern Phenotype Phosphorylation Phosphotransferases Protein Precursors Proteins Publishing Research Retinal Diseases Serum Site Tertiary Protein Structure Testing Therapeutic Therapeutic Studies Toxic effect Transmembrane Domain Variant adeno-associated viral vector design disorder control enzyme deficiency enzyme replacement therapy experimental study gene therapy glycosylation humane endpoint improved lysosomal proteins male man mannose 6 phosphate mortality mouse model nervous system disorder notch protein novel pre-clinical assessment restoration safety assessment sex site-1 protease therapy development trafficking

项目摘要

PROJECT SUMMARY/ABSTRACT Mucolipidosis is a group of inherited conditions in which lysosomal enzymes do not properly traffic to the lysosomes, due to deficient activity of UDP-N-acetylglucosamine: lysosomal enzyme N-acetyl-glucosamine-1- phosphotransferase and subsequent failure to add mannose-6-phosphate residues to lysosomal enzymes. As a result, lysosomes accumulate a variety of substrates that cannot be properly degraded, causing progressive physical and neurological disease and early death. Meanwhile, soluble lysosomal enzymes, lacking mannose- 6-phosphate, can be found in the bloodstream in abnormally high amounts. There is no available treatment. Here, using a null mouse model of mucolipidosis type II (also known as “I-cell disease”), we propose to study gene therapy with a novel S1S3 phosphotransferase for correction of this disorder. We will employ an adeno- associated viral vector, AAV-9, that has the ability to effect widespread transduction of systemic organs and brain when administered intravenously to mice. The experiments are designed to address the following regarding the amenability of mucolipidosis type II to treatment with S1S3 phosphotransferase: 1) biodistribution of phosphotransferase expression to relevant organs including brain, 2) restoration of mannose-6- phosphorylation of lysosomal hydrolases, 3) improvement in lysosomal storage, and the distribution of this effect, 4) improvement in phenotype, including growth and behavior, and 5) lack of apparent toxicity, including mortality, in treated mice. The project combines complementary expertise in glycosylation and trafficking of lysosomal proteins, therapy development for lysosomal diseases, and characterization of the mucolipidosis-II mouse including behavior and pathology.

项目概要/摘要粘脂沉积症是一组遗传性疾病，其中溶酶体酶不能正确运输到溶酶体，由于 UDP-N-乙酰氨基葡萄糖活性不足：溶酶体酶 N-乙酰氨基葡萄糖-1- 磷酸转移酶和随后未能将 6-磷酸甘露糖残基添加到溶酶体酶 As 中。结果，溶酶体积累了多种无法正常降解的底物，导致进行性身体和神经系统疾病和早期死亡同时，可溶性溶酶体酶缺乏甘露糖。 6-磷酸盐在血液中含量异常高，目前尚无可用的治疗方法。在这里，我们建议使用 II 型粘脂沉积症（也称为“I 细胞疾病”）的无效小鼠模型来研究使用新型 S1S3 磷酸转移酶进行基因治疗来纠正这种疾病，我们将采用腺-。相关病毒载体 AAV-9，能够影响全身器官的广泛转导静脉注射给小鼠时的大脑实验旨在解决以下问题。关于 II 型粘脂沉积症对 S1S3 磷酸转移酶治疗的适应性：1) 生物分布包括脑在内的相关器官的磷酸转移酶表达的影响，2) 甘露糖-6-的恢复溶酶体水解酶的磷酸化，3）溶酶体储存的改善及其分布效应，4) 表型改善，包括生长和行为，以及 5) 缺乏明显毒性，包括该项目结合了糖基化和转运方面的互补专业知识。溶酶体蛋白、溶酶体疾病的治疗开发以及粘脂沉积症-II 的特征小鼠，包括行为和病理学。