Novel Reporters and Delivery Vehicles

小说记者和送货车辆

• 批准号: 7729452
• 负责人: XANDRA OWENS BREAKEFIELD
• 金额: $ 15.03万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7729452
• 关键词: Address; Adult; Amino Acid Sequence; Amino Acids; Apoptosis; Arts; Avidin; Binding; Biochemical; Bioluminescence; Biotin; Blood; Brain; Brain Neoplasms; Capsid; Catalytic Domain; Cell Line; Cell Separation; Cell Surface Proteins; Cell surface; Cells; Chemistry; Chimeric Proteins; Cleaved cell; Collaborations; Condition; Cultured Cells; Cultured Tumor Cells; Data; Dependovirus; Detection; Development; Diagnosis; Diagnostic; Diphtheria Toxin; Dose; Effectiveness; Endopeptidases; Endothelial Cells; Endothelium; Engineering; Environment; Estrogen Receptors; Evaluation; Extracellular Matrix; Fibroblast Growth Factor; Firefly Luciferases; Fluorescence; Gelatinase A; Gene Delivery; Generations; Genes; Genetic; Glioblastoma; Glioma; Green Fluorescent Proteins; Growth; Human; Hypoxia; Image; Implant; Improve Access; Infusion procedures; Injection of therapeutic agent; Label; Lentivirus Vector; Libraries; Ligase; Link; Location; Luciferases; LysoTracker; Magnetic Resonance Imaging; Matrix Metalloproteinases; Mediating; Metalloproteases; Methods; Modality; Modeling; Monitor; Mus; Nervous System Neoplasms; Normal tissue morphology; Nude Mice; Numbers; Organ; PECAM1 gene; Peptide Hydrolases; Peptide Library; Peptide Sequence Determination; Peptides; Phenotype; Pilot Projects; Platelet-Derived Growth Factor Receptor; Population; Procedures; Property; Proteins; Recombinant Proteins; Recombinants; Relative (related person); Renilla Luciferases; Reporter; Reporting; Research Design; Reverse Transcriptase Polymerase Chain Reaction; Screening procedure; Sensitivity and Specificity; Signal Transduction; Sorting - Cell Movement; Specificity; Streptavidin; TNF-related apoptosis-inducing ligand; Tail; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Time; Vascular Endothelial Cell; Vascular Endothelial Growth Factor Receptor; Veins; Ventricular; Viral Vector; Virus; Work; adeno-associated viral vector; angiogenesis; base; brain tissue; cell type; cellular transduction; coelenterazine; design; enhanced green fluorescent protein; extracellular; genetically modified cells; imaging probe; immunocytochemistry; improved; in vivo; intravenous administration; killings; macromolecule; mature animal; neoplastic cell; neuropathology; novel; novel therapeutics; particle; promoter; protein expression; receptor; response; sensor; subcutaneous; therapeutic gene; therapeutic protein; tomography; transduction efficiency; tumor; tumor growth; vector; vector genome

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.

因此，迫切需要提高我们对胶质瘤的诊断能力和治疗水平。这个项目 结合了专门针对这些需求的新型遗传和成像方法。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大脑中的肿瘤， 它们可以被遗传修饰以递送新的治疗性蛋白质，包括分泌形式的 凋亡诱导蛋白S-TRAIL，杀死肿瘤细胞，以及血管内皮细胞的诱饵受体。 内皮细胞生长因子（VEGF）受体阻断血管生成。目标2将侧重于发展 生物分子传感器来报告肿瘤环境。第一个传感器将是一个代谢 生物素化的细胞表面标记物，由细胞在对血管生成或缺氧应答的启动子下合成 肿瘤中的信号第二个传感器将是笼状形式的Gaussia荧光素酶，其通过侧翼连接而猝灭。 蛋白质序列，其可以被细胞外肿瘤基质中的金属蛋白酶选择性切割， 从而恢复荧光素酶活性。目的3将鉴定在AAV衣壳上展示的肽，其可以 介导颗粒穿过血液肿瘤屏障。这将涉及新的AAV的体内选择， 肽展示文库，设计用于从肿瘤中分离具有特定表型的感染细胞。靶向 载体将被测试用于改进生物分子传感器和治疗蛋白质以及肽的递送 将评估其他大分子向肿瘤的递送。这些研究结合了联合收割机“设计师” 重组蛋白、遗传修饰的细胞和病毒载体， 生物发光、荧光和MR成像，以监测使用人类肿瘤细胞的递送和响应 植入裸鼠的外周和大脑。我们假设这些方法将允许更多的 中枢神经系统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