Digital Optical Imaging of Vascular Gene Therapy

描述（来自申请人摘要的逐字记录）：动脉粥样硬化 在美国，心血管疾病是死亡的主要原因。 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），已经显示 可用于预防急性血栓形成，阻断血管成形术后再狭窄， 并刺激新血管的生长。然而，目前，没有在 用于精确监测靶细胞内基因表达的体内方法 动脉粥样硬化斑块绿色荧光蛋白的新发现 (GFP)一种标记基因，使完整的活细胞的使用成为可能， 生物作为实验系统。GFP已被广泛用作敏感的 报告基因和从GFP发射的荧光信号可以通过 光学成像本提案将研究使用创新的 数字光学相机，在医学光学成像中心开发， 约翰霍普金斯大学，以检测从GFP发出的光信号。我们将 测试数字光学成像可用于监测的假设 血管基因治疗和血管的体外荧光显微成像 GFP可以扩展到血管GFP的体内数字光学成像。到 为了验证这一假设，我们将开发一种新VEGF-GFP-慢病毒构建体 同时表达VEGF（治疗再狭窄）和GFP（跟踪 VEGF表达。此外，我们还将开发 两种新型的体外和血管内数字光学成像仪， 分子成像工具来监测血管基因治疗，并验证 导致动脉粥样硬化的动物模型。由于目前的知识 基因治疗的生物分布和/或体内药代动力学是不完全的 并且主要依赖于活检或死后组织的染色， 创新成像技术将提供一个独特的机会， 在体内控制血管基因治疗，从而改善 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.