VIRAL VECTORS FOR TARGETED ANTI ANIOGENIC GENE THERAPY

用于靶向抗血管生成基因治疗的病毒载体

• 批准号: 6132525
• 负责人: TAKESHI SANO
• 金额: $ 17.4万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6132525
• 关键词: Retroviridae; angiostatins; epidermal growth factor; gene therapy; glioma; growth factor receptors; immunoconjugates; monoclonal antibody; neoplasm /cancer therapy; tissue /cell culture; transfection /expression vector

DESCRIPTION: (Applicant's Description) The need for repeated administration of anti-factors for long-term therapy of cancer makes gene therapy approaches particularly attractive because sustained expression of anti-angiogenic factors at the tumor site could be achieved. The ability to deliver the anti-angiogenic genes specifically and efficiently to the tumor site will be a key issue for future in vivo anti-angiogenic gene therapy. This exploratory project (R21) aims to design and produce, by using chemical modifications as primary means, retroviral gene transfer vectors that have specific infectivity for target tumor cells for in vivo anti-angiogenic gene therapy of cancer. We will use the retroviral gene transfer vectors, consisting of the viral core of spleen necrosis virus (SNV) and the SNV envelope glycoprotein, which have no infectivity for human cells. However, once these retroviral vectors are made capable of binding specifically to the surface of the target human cell, they can mediate efficient infection by using their natural infection capability. We will use a chemical approach, taking advantage of the extremely tight affinity between streptavidin and biotin, to attach a tumor-specific binding reagent to the surface of retroviral gene transfer vectors such that a tumor-specific infectivity can be generated. Glioma cells that over-express the human epidermal growth factor receptor (hEGFR) and monoclonal antibodies against the extracellular domain of hEGFR will be used as targets and binding mediators, respectively, to see if such modified retroviral gene transfer vectors can infect hEGFR-expressing cells specifically and efficiently. In particular, we will characterize, quantitatively, the degree of modification of the retroviral surface and how each of these modifications affects the binding specificity and infectivity of the modified retroviral gene transfer vectors for the glioma cells over- expressing hEGFR. We will then apply this strategy to SNV-based retroviral gene transfer vectors carrying the angiostatin and endostatin cDNA's to see if these genes can be delivered specifically to the target tumor cells over- expressing hEGFR and if they can be expressed efficiently in the tumor cells.

描述：（申请人的描述） 长期治疗需要重复施用抗因子 癌症使基因治疗方法特别有吸引力，因为持续的 可以实现抗血管生成因子在肿瘤部位的表达。 特异性且有效地递送抗血管生成基因的能力 肿瘤部位将是未来体内抗血管生成基因的关键问题 治疗。 这个探索性项目（R21）旨在设计和生产，通过使用 以化学修饰为主要手段，逆转录病毒基因转移载体 对靶肿瘤细胞具有特异性感染性，可用于体内抗血管生成 癌症的基因治疗。 我们将使用逆转录病毒基因转移载体， 由脾坏死病毒 (SNV) 的病毒核心和 SNV 组成 包膜糖蛋白，对人体细胞无感染性。 然而， 一旦这些逆转录病毒载体能够特异性结合 目标人体细胞表面，它们可以通过以下方式介导有效的感染 利用它们的天然感染能力。 我们将使用化学方法， 利用链霉亲和素和 生物素，将肿瘤特异性结合试剂附着在肿瘤表面 逆转录病毒基因转移载体，从而可以产生肿瘤特异性感染性 生成的。 过度表达人类表皮生长因子的神经胶质瘤细胞 受体（hEGFR）和针对胞外域的单克隆抗体 hEGFR将分别用作靶点和结合介质，看看是否 这种修饰的逆转录病毒基因转移载体可以感染表达 hEGFR 的 细胞特异且有效。 特别是，我们将描述， 定量地，逆转录病毒表面的修饰程度以及如何 这些修饰中的每一个都会影响结合特异性和感染性 用于神经胶质瘤细胞的改良逆转录病毒基因转移载体 表达 hEGFR。 然后我们将将此策略应用于基于 SNV 的逆转录病毒 携带血管抑制素和内皮抑制素 cDNA 的基因转移载体，看看是否 这些基因可以特异性地递送至靶肿瘤细胞 表达 hEGFR 以及它们是否可以在肿瘤细胞中有效表达。