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: 6132955
• 负责人: TERRY A VAN DYKE
• 金额: $ 34.97万
• 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 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6132955
• 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，在此期间，新血管滋养生长的肿瘤并促进肿瘤快速扩张。 最近，Folkman实验室（哈佛）从小鼠中的人肿瘤异种移植物中分离出两种血管生成抑制剂（AngIs），血管抑素和内皮抑素。这些蛋白质抑制内皮细胞增殖，并且当皮下引入小鼠时是异种移植肿瘤生长的有效抑制剂。 这些肿瘤似乎不会对这些因素产生耐药性，这表明它们可以提供有效的长期癌症治疗。 然而，需要更广泛的临床前实验来解决几个重要问题。例如，异种移植模型代表人工实验系统，因为细胞系用于小鼠皮下肿瘤。 此类肿瘤的治疗可能无法反映自然部位新发肿瘤的反应。 在肿瘤从头出现的遗传改变小鼠中进行评估是测试这种方法有效性的下一步。 另一个问题是难以分离出足够的活性因子，甚至用于小鼠研究，这使得人类临床研究具有挑战性。编码这些因子的基因的体细胞递送将避免产生大量纯化蛋白的需要。 该提议旨在测试腺相关病毒（AAV）和腺病毒（ad）载体将体细胞基因递送到良好表征的从头转基因小鼠肿瘤模型中的功效。 该方法还将扩展到至少一个额外的AngI，一种活性血小板反应蛋白-1片段。 具体目标是：（1）表征并优化携带AngI基因的重组AAV和ad载体（rAAV）向小鼠肌肉和肝脏的体细胞递送，以分泌到血清中，以及向脑和乳腺组织的体细胞递送;（2）在充分表征的脉络丛脑肿瘤模型中测试使用AngI的基因疗法（GT）的治疗功效，所述脉络丛脑肿瘤模型经历一致的向血管生成的转变;（3）使用与目的2中的方法类似的方法在非CNS转移性乳腺肿瘤模型中测试AngI GT。