Gene therapy for glycogen storage disease type III

III 型糖原累积病的基因治疗

基本信息

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
项目类别：
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

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

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项目摘要

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.

抽象的糖原储存疾病III型（GSD III）是由缺陷引起的常染色体隐性遗传疾病糖原脱支酶（GDE）导致异常糖原的积累过多（极限在肌肉和肝组织中葡萄蛋白）。大多数患者（约85％）均具有肌肉和肝脏的参与（GSD IIIA）而其他疾病仅限于肝脏（GSD IIIB）。在没有有效疗法的情况下，患者随着GSD III，正在经历渐进性肝衰竭和肌肉损害，伴随着发病率的增加和死亡率。腺相关病毒（AAV）介导的基因疗法已显示出有望治疗遗传的肌肉和肝脏疾病成功翻译成临床试验。但是，这种方法不是 GSD III先进，因为AAV无法传递大型（4.6 kb）人类GDE cDNA 小包装能力。我们已经在小鼠中使用AAV9开发了一种创新的基因治疗方法具有两个关键组件的GSD IIIA模型结合了1）一个小细菌GDE类似物来克服限制小型AAV承载能力； 2）一种新型的免疫凝固双启动子，以防止细胞毒性T 淋巴细胞（CTL）对细菌酶的反应，并在所有受影响组织。该项目的总体目的是确定这一有希望的临床翻译的前进道路治疗。众所周知，鼠标模型中AAV矢量的治疗结果并不总是转化为人类。当前的AAV血清型，尤其是AAV9，将肌肉和肝脏转导高的小鼠效率;但是，在人类患者中转导骨骼肌所需的高剂量AAV9可能导致不良肝毒性甚至肝衰竭。在此提案中，我们旨在确定主要的治疗候选AAV GSD IIIA小鼠中使用高效力跨物种兼容AAV CAPSIDS（CCAAVS）的载体，其中包含DE- 免疫化的转基因表达盒，以最大程度地减少基因治疗相关的免疫反应并减少有效的向量剂量（AIM 1A）。我们将验证GSD IIIA患者肌肉细胞和人类中的铅AAV载体肝脏嵌合小鼠增加其临床转换性（AIM 1B）。然后，我们将检查 GSD IIIA小鼠中的铅AAV载体（AIM 2），并测试其在GSD IIIA犬中的安全性和功效（AIM 3）。数据拟议的研究产生的将奠定将这种创新基因治疗转化为患者的基础与GSD III。使用细菌酶通过基因疗法治疗人类疾病的概念可能会开放为大基因缺陷引起的代谢性疾病的治疗发展的新替代方法。这免疫化双启动子技术也可以广泛用于治疗其他影响的疾病具有基因疗法的多个组织。