Digital Optical Imaging of Vascular Gene Therapy

描述(逐字摘自申请人摘要)：动脉粥样硬化 心血管疾病是美国人死亡的主要原因。 基因治疗是一个迅速发展的领域，具有巨大的应用潜力。 动脉粥样硬化性心血管疾病的治疗。几个特定的基因， 如血管内皮生长因子(phVEGF‘65)，已被证明是 有助于预防急性血栓形成，阻止血管成形术后再狭窄， 并刺激新血管的生长。然而，目前还没有在 一种精确监测靶基因表达的活体方法 动脉粥样硬化斑块。绿色荧光蛋白的最新发现 (GFP)，一种标记基因，使使用完整的活细胞和 生物体作为实验系统。绿色荧光蛋白已被广泛用作一种敏感剂 报告和绿色荧光蛋白发出的荧光信号可以通过 光学成像。这项提案将调查使用创新的 数字光学相机，由医学光学成像中心开发，地址为 约翰霍普金斯大学，探测绿色荧光蛋白发出的光信号。我们会 验证数字光学成像可用于监控的假设 血管基因治疗和体外血管荧光显微成像 GFP可以扩展到血管GFP的在体数字光学成像。至 验证这一假设，我们将开发一种新型的VEGF-GFP-慢病毒构建 同时表达VEGF(用于治疗再狭窄)和GFP(用于跟踪) 血管内皮细胞生长因子表达)。此外，我们还将发展 两种新型的体外和血管内数字光学成像仪 用于监测血管基因治疗的分子成像工具，并验证 在动脉粥样硬化的动物模型中的结果。因为目前关于这方面的知识 基因治疗的生物分布和/或体内药代动力学不完整 主要依靠活组织或死后组织的染色，这 创新的成像技术将提供一个独特的机会来监控和 控制体内血管基因治疗，从而改善对血管的管理 动脉粥样硬化性心血管疾病。

DESCRIPTION (Verbatim from the Applicant's Abstract): Atherosclerotic cardiovascular disease is the leading cause of mortality in the United States. Gene therapy is a rapidly expanding field with great potential for the treatment of atherosclerotic cardiovascular disease. Several specific genes, such as vascular endothelial growth factor (phVEGF'65), have been shown to be useful for preventing acute thrombosis, blocking post-angioplasty restenosis, and stimulating growth of new blood vessels. However, currently, there is no in vivo method for precise monitoring of gene expression within targeted atherosclerotic plaques. The recent discovery of green fluorescent protein (GFP), a marker gene, makes possible the use of intact living cells and organisms as experimental systems. GFP has been widely used as a sensitive reporter and the fluorescence signal emitted from GFP can be detected by optical imaging. This proposal will investigate the use of an innovative digital optical camera, developed at the Center for Medical Optical Imaging at Johns Hopkins University, to detect light signals emitted from GFP. We will test the hypothesis that digital optical imaging can be used to monitor vascular gene therapy, and in vitro fluorescent microscopic imaging of vascular GFPs may be extended to in vivo digital optical imaging of vascular GFPs. To test this hypothesis, we will develop a novel VEGF-GFP-lentiviral construct that expresses simultaneously both VEGF (to treat restenosis) and GFP (to track VEGF expression) after vascular gene transfection. In addition, we will develop two novel, extemal and intravascular digital optical imagers as in vivo molecular imaging tools to monitor vascular gene therapy, and validate the results in animal models of atherosclerosis. Since current knowledge about the biodistribution and/or in vivo pharmacokinetics of gene therapy is incomplete and relies primarily on staining of biopsied or post-mortem tissues, this innovative imaging technique will provide a unique opportunity to monitor and control vascular gene therapy in vivo and thus improve the management of atherosclerotic cardiovascular disorders.