Surgical Approaches to Systemic Gene Transfer

系统性基因转移的外科方法

• 批准号: 6931966
• 负责人: HANSELL H STEDMAN
• 金额: $ 37.64万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6931966
• 关键词: adeno associated virus group; biotechnology; dogs; dystrophin; gene delivery system; gene therapy; hamsters; histamine; immunopathology; muscular dystrophy; nonhuman therapy evaluation; transfection /expression vector; vascular endothelium permeability

DESCRIPTION (provided by applicant): The overall aim of the proposed research is to improve the prospects for therapeutic gene transfer in Duchenne muscular dystrophy by addressing two essential rate-limiting issues: immunity to the transgene product and vector delivery. Using a newly described canine animal model for Duchenne muscular dystrophy, the German Short Haired Pointer, the experimental design takes advantage of a deletion of the dystrophin gene to evaluate the comparative immunogenicity of dystrophin and utrophin. We make exclusive use of rAAV vectors. The experimental design tests the hypothesis that in the context of the deletion, recombinant (canine) mini-dystrophin will elicit a deleterious cellular immune response. It further tests the hypothesis that substitution of a similarly designed canine mini-utrophin transgene will circumvent this immune response. Based on extensive preliminary data, the proposal also addresses the hypothesis that the endothelial barrier to systemic gene delivery can be bypassed by temporarily infusing histamine during a period of mechanical circulatory support. We propose a graded series of experiments to address the latter hypothesis, starting with isolated limb perfusion and culminating in systemic gene delivery. These studies will also make extensive use of another naturally occurring animal model, the hamster model for limb-girdle muscular dystrophy. Successful completion of the experimental plan will provide general information relevant to the immunological response to somatic gene delivery and the preservation of organ function during profound but rapidly reversible alterations in endothelial integrity. It will also provide specific information about the rational design of strategies for systemic gene therapy in one of the most common single-gene lethal diseases in man, Duchenne Muscular Dystrophy.

描述（由申请人提供）：拟议研究的总体目的是通过解决两个基本限制性问题：对转基因产品和矢量输送的免疫力来改善Duchenne肌肉营养不良中治疗基因转移的前景。 使用新描述的犬动物模型用于德国短头发指针Duchenne肌肉营养不良，实验设计利用了肌营养不良蛋白基因的缺失来评估肌营养不良蛋白和乌特罗蛋白的比较免疫原性。我们独家使用Raav载体。实验设计检验了以下假设：在缺失的背景下，重组（犬）迷你脱节蛋白将引起有害的细胞免疫反应。它进一步检验了以下假设：替代类似设计的犬微营养不良蛋白转基因将规避这种免疫反应。基于广泛的初步数据，该提案还解决了以下假设：在机械循环支持的时期，可以通过暂时注入组胺来绕过全身基因递送的内皮屏障。我们提出了一系列分级的实验，以解决后一种假设，从孤立的肢体灌注开始，并在全身基因递送中达到顶点。这些研究还将广泛使用另一种天然发生的动物模型，即肢体肌肉营养不良的仓鼠模型。 成功完成实验计划将提供与对体细胞基因递送的免疫学反应有关的一般信息，并在内皮完整性的深刻但迅速可逆的变化中保存器官功能。它还将提供有关人类Duchenne肌营养不良症中最常见的单基因致命疾病之一的全身基因治疗策略的合理设计。