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

• 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
• 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

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.

项目总结/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地运输到 溶酶体，由于UDP-N-乙酰葡糖胺活性不足：溶酶体酶N-乙酰葡糖胺-1- 磷酸转移酶和随后未能将甘露糖-6-磷酸残基添加到溶酶体酶。作为 结果，溶酶体积累了各种不能被适当降解的底物，导致进行性的 身体和神经系统疾病以及过早死亡。与此同时，缺乏甘露糖的可溶性溶酶体酶- 6-磷酸盐，可以在血液中发现异常高的量。没有可用的治疗方法。 在此，我们使用II型粘脂沉积症（也称为“I-细胞病”）的无效小鼠模型， 用新的S1 S3磷酸转移酶进行基因治疗以纠正这种疾病。我们会用腺- 相关病毒载体AAV-9，其具有影响全身器官的广泛转导的能力， 当静脉注射给小鼠时，实验旨在解决以下问题 关于II型粘脂沉积症对S1 S3磷酸转移酶治疗的适应性：1）生物分布 磷酸转移酶表达到相关器官，包括脑，2）甘露糖-6-磷酸转移酶表达的恢复。 溶酶体水解酶的磷酸化，3）溶酶体储存的改善，以及这种蛋白质的分布。 效果，4）表型改善，包括生长和行为，和5）缺乏明显的毒性，包括 死亡率。该项目结合了糖基化和贩运 溶酶体蛋白、溶酶体疾病的治疗进展和粘脂过多症-II的特征 小鼠包括行为和病理学。