New Vectors and Delivery Systems for Glioblastoma Therapy

胶质母细胞瘤治疗的新载体和递送系统

• 批准号: 7038133
• 负责人: XANDRA OWENS BREAKEFIELD
• 金额: $ 23.06万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-12-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7038133
• 关键词: Alphaherpesvirinae; Epstein Barr virus; Retroviridae; adeno associated virus group; antineoplastics; athymic mouse; carcinogenesis; cell line; cyclophosphamide; fibroblasts; gene therapy; glioma; human genetic material tag; irinotecan; laboratory mouse; neoplasm /cancer therapy; neoplastic cell; prodrugs; recombinant virus; technology /technique development; tissue /cell culture; transfection /expression vector

This project will focus on expanding the therapeutic range of HSV-1 vectors for treatment of brain tumors and on creating zones of resistance to tumor growth in the normal brain by developing novel vectors and delivery modalities. The current proposal plans to increase the therapeutic capacity of the oncolytic HSV-1 vector, MGH2 in two ways: by enhancing infection of glioblastoma (GBM) cells through targeting surface antigens by virions; and by combining oncolytic vectors with amplifying/integrating HSV/AAV amplicon vectors to increase and sustain therapeutic transgene levels. We will also attempt to create a zone of resistance to tumor growth in normal brain via AAV vectors and neuroprecursor cells (MFCs) expressing secretable therapeutic proteins. NPCs and chimeric immune receptor (CIR) lymphocytes will be used to target invasive tumor cells. All studies will be carried out in culture and in intracranial human glioma models in nude mice using bioluminescence imaging reporters to track vector and protein delivery, cell fate and tumor growth. An immunocompetent syngeneic rat GBM model will also be used in evaluating CIR lymphocytes. Aim 1 will explore selective infection of glioblastoma cells expressing the mutant EGF receptor, EGFRvIII, by modification of the glycoprotein C envelope protein of HSV-1 virions for presentation of antibodies to this receptor. Aim 2 will evaluate the ability of CIR lymphocytes to target to EGFRvIII and VEGFR receptors in tumor foci. In Aim 3, AAV vectors and genetically modified NPCs will be used to deliver secretable forms of the tumorspecific apoptotic protein, TRAIL, and the anti-angiogenic factor, Flkl to normal brain to create a region of resistance to tumor growth. Aim 4 will employ a dual gene delivery system combining hybrid amplicon vectors incorporating p5 and ITR DNA elements from AAV, and Rep 78/68 proteins as fusions with the HSV-1 virion tegument protein, VP16. Co-infection with oncolytic HSV-1 or a subset of HSV-1 genes will be evaluated for replicative amplification and genomic integration of amplicon-encoded transgenes. This project will use expertise on oncolytic virus (Project 1, Chiocca), in vivo imaging (Project 3, Weissleder), biostatistics (Core A, Finkelstein), HSV vector stocks (Core B, Krisky/Glorioso), and human GBM cells and neuropathology (Core C, Louis). These studies are designed to complement and increase the therapeutic impact of the HSV-1 oncolytic vector strategy for cancer treatment and to explore the possibility of creating regions of the brain resistant to tumor growth by sustained release of therapeutic proteins, using means deemed compatible with human trials.

该项目将着重于扩展HSV-1载体的治疗范围，用于治疗脑肿瘤以及 通过开发新颖的向量和递送方式，从而产生对正常大脑肿瘤生长的抗性区域。 当前的提案计划通过两种方式提高溶瘤HSV-1载体的治疗能力，MGH2： 通过病毒体靶向表面抗原，增强胶质母细胞瘤（GBM）细胞的感染；并结合 通过放大/整合HSV/AAV扩增器向量增加和维持治疗转基因的Oncolytic向量 水平。我们还将尝试通过AAV向量和 表达可分泌的治疗蛋白的神经前锋细胞（MFC）。 NPC和嵌合免疫受体（CIR） 淋巴细胞将用于靶向侵入性肿瘤细胞。所有研究将在培养和颅内进行 使用生物发光成像记者跟踪载体和蛋白质递送的裸鼠中的人神经瘤模型 命运和肿瘤生长。免疫能力的合成大鼠GBM模型也将用于评估CIR 淋巴细胞。 AIM 1将探索表达突变EGF受体的胶质母细胞瘤细胞的选择性感染， EGFRVIII，通过修饰HSV-1病毒体的糖蛋白C包膜蛋白，以表现为抗体 这个受体。 AIM 2将评估CIR淋巴细胞靶向EGFRVIII和VEGFR受体的能力 焦点。在AIM 3中，AAV矢量和转基因的NPC将用于提供可分泌的特定于特异性的形式 凋亡蛋白，小径和抗血管生成因子，FLKL到正常大脑，以创建抗性区域 肿瘤生长。 AIM 4将采用一个双基因输送系统，结合了融合P5的混合动力扩增子向量 来自AAV的ITR DNA元素，REP 78/68蛋白作为与HSV-1病毒素Tegument蛋白的融合，VP16。 将评估与溶瘤HSV-1或HSV-1基因子集的共感染，以进行复制扩增和 扩增子编码转基因的基因组整合。该项目将在溶瘤病毒上使用专业知识（项目1， Chiocca），体内成像（项目3，Weissleder），生物统计学（核心A，Finkelstein），HSV矢量库存（核心B，核心B， krisky/glorioso）和人类GBM细胞和神经病理学（核心C，路易斯）。这些研究被设计为 补充并增加HSV-1肿瘤载体策略对癌症治疗的治疗影响 探索通过持续释放治疗的脑部抗肿瘤生长区域的可能性 蛋白质，使用与人类试验兼容的手段。