Modulation of Angiogenesis Via the Angiostatin Receptor

描述：（申请人的摘要）该项目最初提交于 对PAR-98-096，疾病中血管生成的治疗调节的反应。在 这个修订后的项目，我们寻求开发抗体和肽， 血管生成通过内皮细胞（EC）血管抑素受体，最近， 我们发现是细胞表面ATP合酶。ATP合成酶在细胞表面 肿瘤EC可能在低能量下的补充能量产生中起作用， 氧条件或参与肿瘤的内皮信号传导事件 血管生成针对ATP合酶亚基的多克隆抗体竞争 与血管抑素进行细胞表面EC结合和阻断的能力 血管抑素抑制EC增殖和迁移。多克隆抗血清 对b亚基的直接EC抑制作用也超过 血管抑制素此外，我们最近发现了第二个ATP生产 EC表面上的酶，核苷二磷酸激酶h1（NDPK h1），即 被血管抑制素显著抑制（提交出版的手稿， 见附录）。这些和其他研究结果支持了血管抑素 通过破坏表面ATP对EC发挥抗增殖作用 合成.血管抑素抑制血管生成的公认能力 体内许多肿瘤的生长和转移也是通过抑制 EC表面ATP合成。然而，血管抑素本身作为一种抗肿瘤药物的潜力很差。 由于在生产、稳定性 和亲和力。我们发现了血管抑素的两个EC靶点， 有可能开发出更强大的化合物，可以阻止血管生成， 人类癌症和其它增殖性疾病。该项目的目标是 开发可以替代血管抑素的抗体和肽， 通过与ATP的直接相互作用在乳腺癌中的血管生成 在EC表面上的合成装置。这些化合物作为 治疗剂证明了密集的努力，以开发适当的 人源化单克隆抗体和靶向肽噬菌体展示文库。 将筛选候选抗体和肽的EC抑制活性 使用已建立的测定增殖、迁移和管 阵此外，我们还开发了一种新的检测EC表面ATP的方法， 将用于筛选化合物的抑制活性的合成。 在任何这些测定中表现出抑制活性的化合物将是 使用人乳腺癌筛选它们在体内抑制肿瘤生长的能力 癌症异种移植模型。由于暴露的血管内定位， ATP合成酶，该项目将产生一个集合， 具有快速转化为抗血管生成化合物的强大潜力， 临床研究。

DESCRIPTION: (Applicant's Abstract) This project was originally submitted in response to PAR-98-096, THERAPEUTIC MODULATION OF ANGIOGENESIS IN DISEASE. In this revised project, we seek to develop antibodies and peptides that modulate angiogenesis through the endothelial cell (EC) angiostatin receptor, recently shown by us to be cell surface ATP synthase. ATP synthase on the surface of tumor EC may play a role both in supplemental energy production under low oxygen conditions or in endothelial signaling events involved in tumor angiogenesis. Polyclonal antibodies against subunits of ATP synthase compete with angiostatin for cell surface EC binding and block the ability of angiostatin to inhibit EC proliferation and migration. Polyclonal antisera against the b-subunit also exhibit a direct EC inhibitory effect exceeding that of angiostatin. Furthermore, we recently discovered a second ATP producing enzyme on the EC surface, nucleoside diphosphate kinase h1 (NDPK h1), that is dramatically inhibited by angiostatin (manuscript submitted for publication, see Appendix). These and other findings support the hypothesis that angiostatin exerts its anti-proliferative effect on EC through disruption of surface ATP synthesis. It is likely that the recognized ability of angiostatin to inhibit growth and metastasis of many tumors in vivo is also mediated by inhibition of EC surface ATP synthesis. However, angiostatin itself has poor potential as a therapeutic agent in humans because of limitations in production, stability, and affinity. Our discovery of two EC targets of angiostatin offers the possibility of developing more robust compounds that can block angiogenesis in human cancer and other proliferative diseases. The goal of this project is to develop antibodies and peptides that can substitute for angiostatin to inhibit angiogenesis in breast cancer through direct interactions with the ATP synthesizing apparatus on the surface of EC. The promise of these compounds as therapeutic agents justifies an intensive effort to develop appropriate humanized monoclonal antibodies and targeted peptide phage display libraries. Candidate antibodies and peptides will be screened for EC inhibitory activity using established assays that measure proliferation, migration, and tube formation. In addition, we have developed a novel assay for EC surface ATP synthesis that will be used to screen compounds for inhibitory activity. Compounds that exhibit inhibitory activity in any of these assays will be screened for their ability to inhibit tumor growth in vivo using a human breast cancer xenograft model. Because of the exposed intravascular localization of the ATP synthesizing enzymes, this project will yield a collection of anti-angiogenic compounds with strong potential for rapid translation into clinical studies.