VIRAL VECTORS FOR TARGETED ANTI ANIOGENIC GENE THERAPY

描述：（申请人的描述） 需要反复给予抗因子以长期治疗 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。 特异性和有效地传递抗血管生成基因的能力 将肿瘤部位的抗血管生成基因转移到体内将是未来抗血管生成基因的关键问题 疗法 该探索性项目（R21）旨在设计和生产，通过使用 化学修饰作为主要手段，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，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 包膜糖蛋白，对人体细胞无感染性。 然而，在这方面， 一旦使这些逆转录病毒载体能够特异性结合 在靶人类细胞的表面，它们可以介导有效的感染， 利用它们的自然感染能力。 我们将使用化学方法， 利用链霉抗生物素蛋白和 生物素，以将肿瘤特异性结合试剂附着到 逆转录病毒基因转移载体，使得可以 生成的. 过度表达人表皮生长因子的胶质瘤细胞 受体（hEGFR）和抗hEGFR细胞外结构域的单克隆抗体 hEGFR将分别用作靶点和结合介质，以观察 这种修饰的逆转录病毒基因转移载体可以感染表达hEGFR的 细胞特异性和有效性。 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 这些修饰中的每一种都会影响 用于胶质瘤细胞的经修饰的逆转录病毒基因转移载体超过- 表达hEGFR。 然后，我们将把这一策略应用于基于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， 这些基因可以特异性地递送到靶肿瘤细胞， 表达hEGFR以及它们是否可以在肿瘤细胞中有效表达。

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.