Gene therapy for glycogen storage disease type III

III 型糖原累积病的基因治疗

• 批准号: 10444983
• 负责人: Baodong Sun
• 金额: $ 70.75万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10444983
• 关键词: Adult; Affect; Age; Age-Months; Bacillus subtilis; Biochemical; Biological Markers; Blood; Canis familiaris; Capsid; Cardiac; Carrying Capacities; Cells; Chickens; Clinical Trials; Complementary DNA; Cytomegalovirus; Cytotoxic T-Lymphocytes; DNA cassette; Data; Defect; Dependovirus; Development; Dextrins; Disease; Dose; Enhancers; Enzymes; Female; Foundations; Gender; Genes; Glycogen; Glycogen Debranching Enzyme; Glycogen Storage Disease Type III; Goals; Hepatocyte; Hepatotoxicity; Hereditary Disease; Histologic; Human; Immune response; Infant; Inherited; Knock-out; Laboratories; Lead; Liver; Liver Cirrhosis; Liver Failure; Liver diseases; Measurement; Mediating; Metabolic Diseases; Morbidity - disease rate; Mus; Muscle; Muscle Cells; Muscle Fibers; Muscle function; Mutation; Myocardium; Myopathy; Oncogenes; Open Reading Frames; Organ; Outcome; Patients; Recording of previous events; Research; Safety; Serotyping; Serum; Skeletal Muscle; Technology; Testing; Tissues; Toxic effect; Translating; Translations; Urine; adaptive immune response; adeno-associated viral vector; aged; analog; beta Actin; canine model; cell type; clinical candidate; clinical translation; clinically translatable; curative treatments; de-immunization; dietary; effective therapy; efficacy evaluation; experience; gene therapy; genome editing; human disease; humanized mouse; innovation; liver function; mortality; mouse model; novel; prevent; promoter; response; sex; skeletal; therapeutic candidate; therapeutic development; therapy development; therapy outcome; transduction efficiency; transgene expression; urinary; vector

ABSTRACT Glycogen storage disease type III (GSD III) is an autosomal recessive inherited disorder caused by deficiency of glycogen debranching enzyme (GDE) that leads to excessive accumulation of abnormal glycogen (limit dextrin) in muscle and liver tissues. The majority of patients (~85%) have both muscle and liver involvement (GSD IIIa) while others have disease limited to the liver (GSD IIIb). In absence of an effective therapy, patients with GSD III are experiencing progressive liver failure and muscle damage accompanied by increased morbidity and mortality. Adeno-associated virus (AAV) mediated gene therapy has shown promise for treating inherited muscle and liver disorders with successful translation to clinical trials. However, this approach has not been advanced for GSD III because AAV is not capable of delivering the large (4.6 kb) human GDE cDNA, due to its small packaging capacity. We have developed an innovative gene therapy approach with AAV9 in a mouse model of GSD IIIa with two key components incorporated 1) a small bacterial GDE analog to overcome the limitation of small AAV carrying capacity; and 2) a novel immunotolerizing dual promoter to prevent cytotoxic T lymphocyte (CTL) response to the bacterial enzyme and enable long-term Pullulanase expression in all affected tissues. The overall objective of this project is to identify a path forward for clinical translation of this promising therapy. It is commonly known that the therapeutic outcomes of AAV vectors in mouse models does not always translate into human. Current AAV serotypes, especially AAV9, transduce muscle and liver in mice with high efficiency; however, high doses of AAV9 required to transduce skeletal muscles in human patients can led to adverse hepatotoxicity or even liver failure. In this proposal, we aim to identify a lead therapeutic candidate AAV vector in GSD IIIa mice using high potency cross-species compatible AAV capsids (ccAAVs) containing a de- immunized transgene expression cassette to minimize gene therapy related immune responses and reduce the effective vector dose (Aim 1a). We will validate the lead AAV vector in GSD IIIa patient muscle cells and human liver chimeric mice to increase its clinical translatability (Aim 1b). We will then examine the long-term efficacy of the lead AAV vector in GSD IIIa mice (Aim 2), and test its safety and efficacy in GSD IIIa dogs (Aim 3). Data generated from the proposed studies will lay the foundation for translating this innovative gene therapy to patients with GSD III. The concept of using a bacterial enzyme to treat human diseases through gene therapy may open up new alternatives for therapeutic development for metabolic disorders caused by defects in large genes. The immunotolerizing dual promoter technology can also be broadly used for treating other conditions that affect multiple tissues with gene therapy.

摘要 糖原累积病III型（GSD III）是一种常染色体隐性遗传性疾病， 糖原脱支酶（GDE），导致异常糖原的过度积累（限制 糊精）在肌肉和肝组织中的含量。大多数患者（~85%）同时累及肌肉和肝脏 (GSD IIIa），而其他人的疾病仅限于肝脏（GSD IIIb）。在缺乏有效治疗的情况下， 患有GSD III的患者正在经历进行性肝功能衰竭和肌肉损伤，并伴有发病率增加 and mortality.腺相关病毒（AAV）介导的基因治疗已显示出治疗遗传性 肌肉和肝脏疾病，并成功转化为临床试验。然而，这种方法并没有被 因为AAV不能递送大的（4.6kb）人GDE cDNA，这是由于其 包装容量小。我们在小鼠中开发了一种创新的AAV 9基因治疗方法， GSD IIIa的模型，其具有两个关键组分：1）小的细菌GDE类似物，以克服 小AAV携带能力的限制;和2）防止细胞毒性T细胞的新型免疫耐受性双重启动子 淋巴细胞（CTL）对细菌酶的反应，并使所有受影响的普鲁兰酶长期表达 组织中该项目的总体目标是确定一条道路，为临床翻译这一有前途的 疗法众所周知，AAV载体在小鼠模型中的治疗结果并不总是 翻译成人类。目前的AAV血清型，特别是AAV 9，在具有高表达的小鼠中感染肌肉和肝脏， 然而，在人类患者中，需要高剂量的AAV 9来刺激骨骼肌，可能导致 不利的肝毒性甚至肝衰竭。在这项提案中，我们的目标是确定一个领先的治疗候选AAV 载体在GSD IIIa小鼠中使用高效力跨物种相容性AAV衣壳（ccAAV），所述AAV衣壳含有去- 免疫的转基因表达盒，以使基因治疗相关的免疫应答最小化，并降低免疫应答。 有效载体剂量（目标1a）。我们将在GSD IIIa患者肌肉细胞和人肌肉细胞中验证前导AAV载体。 肝嵌合小鼠，以增加其临床可翻译性（目的1b）。然后我们将研究 在GSD IIIa小鼠中的前导AAV载体（目的2），并在GSD IIIa狗中测试其安全性和有效性（目的3）。数据 从拟议的研究中产生的结果将为将这种创新的基因疗法转化为患者奠定基础 关于GSD III利用细菌酶通过基因疗法治疗人类疾病的概念可能会开启 为大基因缺陷引起的代谢紊乱的治疗开发提供新的替代方案。的 免疫耐受化双启动子技术也可广泛用于治疗其它影响 多个组织进行基因治疗