VIRAL VECTORS FOR TARGETED ANTI ANIOGENIC GENE THERAPY

描述：(申请人描述) 长期治疗需要反复应用抗因子药物 癌症使基因治疗方法特别吸引人，因为它具有持续性 可以在肿瘤部位表达抗血管生成因子。 特异性和高效地传递抗血管生成基因的能力 肿瘤部位的定位将是未来体内抗血管生成基因研究的关键问题 心理治疗。该探索性项目(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 对体内抗血管生成的靶肿瘤细胞具有特异性感染性 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与 靶细胞表面，它们可以通过以下途径介导有效的感染 利用它们的自然感染能力。我们将使用化学方法， 利用链霉亲和素和链霉亲和素之间极其紧密的亲和力 生物素，将肿瘤特异性结合试剂附着在 逆转录病毒基因转移载体，使肿瘤特异性感染性 已生成。过表达人表皮生长因子的胶质瘤细胞 人表皮生长因子受体(HEGFR)和抗人球蛋白胞外区的单抗 HEGFR将分别被用作靶标和结合介体，以查看是否 这种修饰的逆转录病毒基因转移载体可以感染表达hEGFR的人 特定而高效的细胞。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 这些修饰中的每一种都会影响结合的特异性和感染性 改良型逆转录病毒基因转移载体对脑胶质瘤细胞的杀伤作用 表达人表皮生长因子受体。然后我们将把这一策略应用于基于SNV的逆转录病毒 携带血管抑素和内皮抑素基因转移载体的研究 这些基因可以通过以下途径特异性地传递到靶肿瘤细胞- HEGFR的表达以及它们是否能在肿瘤细胞中高效表达。

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.