Gene delivery to striated muscle by systemic AAV vectors

通过系统性 AAV 载体将基因递送至横纹肌

• 批准号: 7901568
• 负责人: Dwight D Koeberl
• 金额: $ 30.3万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7901568
• 关键词: Acids; Affect; Age of Onset; Alpha-glucosidase; Antibodies; Antigens; Biological Assay; Biological Markers; Capsid; Cell physiology; Cells; Cessation of life; Cytoplasm; Data; Dependovirus; Detection; Disease; Enzymes; Failure; Figs - dietary; Gene Delivery; Glucan 1,4-alpha-Glucosidase; Glycogen; Glycogen Storage Disease; Glycogen storage disease type II; Heart; Humoral Immunities; Hypertrophic Cardiomyopathy; IGF Type 2 Receptor; Immune response; Immunocompetent; Infusion procedures; Knock-out; Knockout Mice; Limb structure; Liver; Lymphocyte; Lysosomes; Measures; Mediating; Molecular; Monitor; Mus; Muscle; Muscle Weakness; Muscle hypotonia; Muscular Dystrophies; Myopathy; Outcome Measure; Patients; Peptide Signal Sequences; Performance; Production; Regulatory T-Lymphocyte; Research Personnel; Respiratory Diaphragm; Role; Serotyping; Severities; Signal Induction; Signal Transduction; Skeletal Muscle; Striated Muscles; T cell response; adeno-associated viral vector; enzyme replacement therapy; gene therapy; glucosidase; in vivo; infancy; intravenous injection; neutralizing antibody; novel strategies; particle; programs; promoter; receptor; research study; uptake; urinary; vector

DESCRIPTION (provided by applicant): Glycogen storage disease II (GSD-II; Pompe disease; MIM 232300) is a lethal muscular dystrophy caused by the deficiency of acid alpha-glucosidase (GAA; acid maltase). No curative therapy is available for GSD-II. Obstacles to gene therapy in GSD-II include very high GAA requirements and the occurence of neutralizing anti-GAA antibodies. Gene therapy with adeno-associated virus (AAV) vectors was advanced by the availability of AAV serotypes 6-9, termed myotrophic AAV vectors in this application. The general hypothesis states that myotrophic AAV vectors will express efficacious GAA in the striated muscles of GSD-II mice, including the heart, diaphragm, and skeletal muscles. Myotrophic AAV2 vectors pseudotyped as AAV7, AAV8, and AAV9 will be delivered intravenously to analyze the systemic delivery of these vectors to striated muscle. Specific Hypothesis 1: Pseudotyped AAV vectors containing a muscle-specific promoter will express high levels of GAA in the striated muscle of GSD-II mice. Correction of GAA deficiency and glycogen storage in the heart, diaphragm, and skeletal muscle will be analyzed for each serotype in GAA- knockout (GAA-KO) mice. Myotrophic AAV vectors will be administered to infantile GAA-KO mice, demonstrating efficacy in Pompe disease. Specific Hypothesis 2: An AAV vector will evade cellular and humoral immune responses to achieve long-term efficacy in GSD-II mice. Cellular and humoral immune responses to hGAA expression restricted to striated muscles will be analyzed. ELISpot assays will be performed to analyze antigen-specific T cell responses to hGAA and AAV capsids, and flow cytometric detection of additional lymphocyte markers will evaluate the immune response to AAV vectors. Specific Hypothesis 3: Liver-specific expression will Induce tolerance to introduced GAA and enhance the efficacy of myotrophic AAV vectors in GSD-II mice. Tolerance to GAA will be induced by liver-specific expression of GAA in immunocompetent GAA-KO mice. T regulatory cells will be quantitated in tolerant GAA-KO mice; furthermore, T regulatory cells will be depleted to demonstrate their role in establishing tolerance to GAA. A novel strategy will deliver an AAV vector encoding chimeric GAA by intravenous injection and isolated limb delivery to convert skeletal muscle to a depot for secreted GAA. Efficacious muscle-targeted gene therapy in GSD-II will have implications for gene therapy in other muscular dystrophies and myopathies.

描述（由申请人提供）： 糖原储存疾病II（GSD-II； Pompe疾病； MIM 232300）是由酸α-葡萄糖苷酶（GAA；酸麦芽糖酶）缺乏引起的致命肌肉营养不良。 GSD-II没有治疗疗法。 GSD-II基因治疗的障碍包括GAA的高需求和中和抗GAA抗体的发生。通过在本应用程序中，AAV血清型6-9的可用性称为AAV血清型，使用腺相关病毒（AAV）载体的基因治疗是通过可用的。一般假设指出，肌营养的AAV载体将在GSD-II小鼠的横纹肌肉中表达有效的GAA，包括心脏，隔膜和骨骼肌。将静脉注射为AAV7，AAV8和AAV9的肌营养AAV2载体将静脉内输送为AAV7，AAV8和AAV9，以分析这些向量的系统性输送到横纹肌肉。特定的假设1：含有肌肉特异性启动子的伪型AAV向量将在GSD-II小鼠的横纹肌中表达高水平的GAA。将分析GAA敲除（GAA-KO）小鼠中每种血清型的心脏，隔膜和骨骼肌中GAA缺乏症和糖原储存的校正。肌营养的AAV载体将用于婴儿GAA-KO小鼠，表现出在庞贝疾病中的功效。特定假设2：AAV载体将逃避细胞和体液免疫反应，以实现GSD-II小鼠的长期疗效。将分析对限制在横纹肌肉的HGAA表达的细胞和体液免疫反应。将进行ELISPOT分析，以分析抗原特异性T细胞对HGAA和AAV CAPSIDS的响应，并且其他淋巴细胞标记物的流式细胞仪检测将评估对AAV载体的免疫反应。特定假设3：肝特异性表达将诱导引入GAA的耐受性，并增强肌营养AAV载体在GSD-II小鼠中的疗效。在免疫能力的GAA-KO小鼠中，GAA的肝特异性表达将诱导对GAA的耐受性。耐受性GAA-KO小鼠将定量调节细胞；此外，T调节细胞将耗尽以证明其在建立对GAA耐受性的作用。一种新颖的策略将通过静脉注射和孤立的肢体递送来提供编码嵌合GAA的AAV矢量，以将骨骼肌转换为分泌GAA的仓库。 GSD-II中有效针对肌肉的基因疗法将对其他肌肉营养不良和肌病中的基因治疗产生影响。