MECHANISMS OF FGF ENHANCED ADENOVIRAL GENE DELIVERY

FGF 增强腺病毒基因传递的机制

基本信息

批准号：
2715811
负责人：
John Doukas
金额：
$ 10万
依托单位：
SELECTIVE GENETICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-09-01 至 1999-06-30
项目状态：
已结题

项目摘要

Gene therapy using adenoviruses is limited by the inability to selectively deliver DNA to specific targets. Retargeting can be accomplished by introducing fibroblast growth factor (FGF) onto viral capsids through its conjugation to neutralizing anti-viral antibodies. As a result, vectors bind to cells not through their native receptors, but rather through FGF receptors overexpressed on tumors and proliferating endothelium. In addition, an increased transduction efficiency is accomplished, which we hypothesize results from the high affinity of FGF receptors (Kd= approximately 2 pM) and their ability to traffic to the nucleus. In this application we propose to examine the mechanisms of FGF retargeting in vitro. Specific aim one will define the roles of FGF and adenoviral receptors in cell surface binding and transduction. Specific aim two will confirm the intracellular pathways of virus internalization and gene delivery. Phase II studies would use this information to rationally develop in vivo models for examining the influence of FGF-retargeting on viral toxicity and immunogenicity, and for the development other ligand-viral conjugates. The data generated will validate the use of growth factor retargeting for gene therapy, and establish the groundwork required for its use in clinical trials. PROPOSED COMMERCIAL APPLICATION: The proposed studies are designed to develop therapeutic products for gene therapy. Initial drug candidates will target adenoviruses to growth factor receptors overexpressed on tumor cells and associated vasculature, while at the same time eliminating normal viral tropism for uninvolved organs. Targeted delivery will allow transduction of tumors with cytotoxic proteins and other anti-proliferative agents, and accomplish antitumor activity by destruction of tumor cells and/or proliferating endothelium. This approach will allow application of the growing number of adenoviral vectors to cancer gene therapy by eliminating their major limitation, namely non-targeted delivery to normal tissues.

使用腺病毒的基因治疗受到无法有选择地将DNA传递到特定靶标。可以重新定位通过将成纤维细胞生长因子（FGF）引入病毒来完成衣壳通过其结合到中和抗病毒抗体的结合。结果，向量与细胞结合而不是通过其天然受体结合，而是通过FGF受体过表达的肿瘤和增生的内皮。另外，转导增加效率是实现的，我们假设这是高度的结果 FGF受体的亲和力（KD =大约2 pm）及其能力核的流量。在此应用中，我们建议检查 FGF在体外重新定位的机制。特定目标将定义 FGF和腺病毒受体在细胞表面结合以及转导。具体目标两个将确认细胞内途径病毒内在化和基因递送。第二阶段研究将使用这些信息以合理开发用于检查的体内模型 FGF重新定位对病毒毒性和免疫原性的影响，以及为了开发其他配体病毒缀合物。生成的数据将验证生长因子重新定位用于基因疗法的使用，并建立在临床试验中使用所需的基础。拟议的商业应用：拟议的研究旨在开发用于基因疗法。最初的候选药物将针对腺病毒在肿瘤细胞上过表达的生长因子受体和相关的受体脉管系统，同时消除了正常病毒性向潮流无参与器官。有针对性的分娩将允许转导肿瘤具有细胞毒性蛋白和其他抗增殖剂，以及通过破坏肿瘤细胞和/或来实现抗肿瘤活性增生的内皮。这种方法将允许应用越来越多的腺病毒载体通过消除其主要限制，即非目标交付正常组织。