Novel Reporters and Delivery Vehicles

我们迫切需要提高我们对胶质瘤的诊断和治疗能力。这个项目 结合了新的基因和成像方法，专门满足了这些需求。在目标1中，我们将探索 成年动物的内源性神经前体细胞是否会迁移到脑肿瘤，以及 它们可以通过基因改造来传递新的治疗性蛋白质，包括分泌形式的 杀伤肿瘤细胞的凋亡诱导蛋白S-TRAIL，以及血管的诱骗受体 血管内皮细胞生长因子受体阻断血管生成。目标2将专注于开发 生物分子传感器从肿瘤环境中报告。第一个传感器将是新陈代谢的 细胞在血管生成或缺氧反应启动子作用下合成的生物素化细胞表面标记 肿瘤中的信号。第二个传感器将是笼形的高斯荧光素酶，通过侧翼淬灭。 蛋白质序列，可被细胞外肿瘤基质中的金属蛋白酶选择性切割， 从而恢复荧光素酶的活性。AIM 3将识别显示在AAV衣壳上的多肽，这可以 调节粒子通过血液-肿瘤屏障。这将涉及到体内选择一种新型的AAV 为从肿瘤中分离特定表型的感染细胞而设计的多肽展示文库。瞄准 将对载体进行测试，以改进生物分子传感器、治疗性蛋白质和多肽的传递 将被评估是否将其他大分子输送到肿瘤。这些研究结合了“设计师” 重组蛋白、转基因细胞和病毒载体 生物发光、荧光和磁共振成像用于监测人体肿瘤细胞的传递和反应 植入裸鼠的外周和脑内。我们假设这些方法将允许更多 中枢神经肿瘤的有效诊断和客观治疗评价，具有广泛的适用性 其他类型的肿瘤。这项工作将与项目4(荧光蛋白)合作进行 断层扫描)和试点项目1(神经前体细胞系)，并将取决于小鼠成像核心和 化学核心。

There is an urgent need to improve our diagnostic capabilities and treatment of glioma tumors. This project combines novel genetic and imaging methods that specifically address these needs. In Aim 1 we will explore whether endogenous neuroprecursor cells in adult animals will migrate to tumors in the brain and whether they can be genetically modified to deliver new therapeutic proteins, including secreted forms of the apoptosis-inducing protein, S-TRAIL, to kill tumor cells, as well as a decoy receptor for the vascular endothelial cell growth factor (VEGF) receptor to block angiogenesis. Aim 2 will focus on development of biomolecular sensors to report from the tumor environment. The first sensor will be a metabolically biotinylated cell surface marker synthesized by cells under promoters that respond to angiogenic or hypoxic signals in the tumor. The second sensor will be a caged form of Gaussia luciferase quenched by flanking protein sequences, which can be selectively cleaved by metalloproteinases in the extracellular tumor matrix, thus restoring luciferase activity. Aim 3 will identify peptides displayed on the AAV capsid, which can mediate passage of particles across the blood-tumor barrier. This will involve in vivo selection of a novel AAV peptide-display library designed to isolate infected cells of specific phenotypes from the tumors. Targeting vectors will be tested for improved delivery of biomolecular sensors and therapeutic proteins, and peptides will be evaluated for delivery of other macromolecules to tumors. These studies combine "designer" recombinant proteins, genetically modified cells, and virus vectors with state-of-the-art methods of bioluminescence, fluorescence and MR imaging to monitor delivery and responses using human tumor cells implanted in the periphery and brains of nude mice. We hypothesize that these approaches will allow more efficient diagnosis and objective treatment evaluation of central nervous tumors, with broad applicability to other types of tumors. This work will be carried out in collaboration with Project 4 (fluorescent protein tomography) and Pilot Project 1 (neuroprecursor cell line), and will depend on the Mouse Imaging Core and Chemistry Core.