ANTI-ANGIOGENESIS GENE THERAPY FOR DE NOVO TUMORS

针对新发肿瘤的抗血管生成基因疗法

• 批准号: 6514545
• 负责人: TERRY A VAN DYKE
• 金额: $ 34.69万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6514545
• 关键词: Adenoviridae; adeno associated virus group; angiogenesis inhibitors; angiostatins; breast neoplasms; gene targeting; gene therapy; genetically modified animals; laboratory mouse; neoplasm /cancer blood supply; neoplastic process; recombinant virus; transfection; transfection /expression vector

Angiogenesis is an essential component of tumor progression during which new blood vessels nourish growing tumors and facilitate rapid tumor expansion. Recently, two angiogenesis inhibitors (AngIs), angiostatin and endostatin, were isolated from human tumor xenografts in mice by the Folkman lab (Harvard). These proteins inhibit endothelial cell proliferation and are effective inhibitors of xenograft tumor growth when introduced into mice subcutaneously. Such tumors do not appear to develop resistance to these factors, suggesting that they could provide effective long-term therapy against cancer. However, more extensive preclinical experimentation is required to address several important issues. For example, the xenograft models represent an artificial experimental system, since cell lines are used to tumors subcutaneously in the mouse. Treatment of such tumors may not reflect the response of de novo tumors arising in natural sites. Assessment in genetically altered mice where tumors arise de novo is the next step in testing the efficacy of this approach. Another problem has been in the difficulty in isolating enough active factor for even the mouse studies, making human clinical studies challenging. Somatic delivery of the genes encoding these factors would circumvent the need to produce large quantities of purified protein. This proposal aims to test the efficacy of somatic gene delivery by adeno-associated virus (AAV) and adenovirus (ad) vectors into well-characterized de novo transgenic mouse tumor models. The approach will also be extended to at least one additional AngI, an active thrombospondin-1 fragment. The specific aims are to: (1) Characterize and optimize somatic delivery of recombinant AAV and ad vectors (rAAV) carrying the AngI genes to mouse muscle and liver for secretion to serum, and to brain and mammary tissue; (2) Test the therapeutic efficacy of genetic therapy (GT) with AngIs in a well characterized choroid plexus brain tumor model that undergoes a consistent transition to angiogenesis; (3) Test AngI GT in a non-CNS metastatic mammary tumor model using approaches similar to those in aim 2.

血管生成是肿瘤进展的重要组成部分，在此过程中，新生血管滋养肿瘤生长并促进肿瘤快速扩张。最近，哈佛大学Folkman实验室从小鼠的人类肿瘤异种移植中分离出两种血管生成抑制剂（AngIs）：血管抑制素和内皮抑制素。这些蛋白能抑制内皮细胞的增殖，并能有效抑制小鼠皮下移植瘤的生长。这些肿瘤似乎没有对这些因素产生耐药性，这表明它们可以提供有效的长期治疗癌症的方法。然而，需要更广泛的临床前实验来解决几个重要问题。例如，异种移植模型代表了一种人工实验系统，因为细胞系用于小鼠皮下肿瘤。这类肿瘤的治疗可能不能反映自然部位新生肿瘤的反应。在重新产生肿瘤的转基因小鼠中进行评估是测试这种方法有效性的下一步。另一个问题是，即使在小鼠研究中也很难分离出足够的活性因子，这给人类临床研究带来了挑战。体细胞递送编码这些因子的基因将避免产生大量纯化蛋白的需要。本研究旨在验证腺相关病毒（AAV）和腺病毒（ad）载体将体细胞基因传递到具有良好特征的新生转基因小鼠肿瘤模型中的有效性。该方法还将扩展到至少一个额外的AngI，一个活跃的血栓反应蛋白-1片段。具体目的是：(1)表征和优化携带AngI基因的重组AAV和ad载体（rAAV）在小鼠肌肉和肝脏的体传递，并将其分泌到血清、脑和乳腺组织；(2)在血管生成过渡的脉络膜丛脑肿瘤模型中检测基因治疗（GT）与AngIs的治疗效果；(3)在非中枢神经系统转移性乳腺肿瘤模型中使用与目的2相似的方法检测AngI GT。