Surgical Approaches to Systemic Gene Transfer

系统性基因转移的外科方法

• 批准号: 6799192
• 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/6799192
• 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）：拟议研究的总体目标是通过解决两个基本限速问题来改善杜氏肌营养不良症治疗性基因转移的前景：对转基因产物的免疫力和载体递送。 使用新描述的杜氏肌营养不良症犬动物模型，德国短毛指针，实验设计利用肌营养不良蛋白基因的缺失来评估肌营养不良蛋白和肌营养不良蛋白的比较免疫原性。我们专门使用rAAV载体。实验设计测试了以下假设：在缺失的情况下，重组（犬）微型肌养蛋白将引发有害的细胞免疫应答。它进一步测试了类似设计的犬微肌营养蛋白转基因的替代将规避这种免疫应答的假设。基于广泛的初步数据，该提案还提出了这样的假设，即在机械循环支持期间，通过暂时输注组胺可以绕过全身基因递送的内皮屏障。我们提出了一个分级系列的实验，以解决后者的假设，孤立的肢体灌注和最终在全身基因传递。这些研究还将广泛使用另一种自然发生的动物模型，即肢带型肌营养不良症的仓鼠模型。 实验计划的成功完成将提供与体细胞基因递送的免疫应答以及在内皮完整性的深刻但快速可逆的改变期间器官功能的保存相关的一般信息。它也将提供有关的系统基因治疗策略的合理设计的具体信息，在一个最常见的单基因致死性疾病的人，杜氏肌营养不良症。