Modulation of Angiogenesis Via the Angiostatin Receptor

描述：(申请者摘要)本项目最初提交于 对PAR-98-096的反应，疾病中血管生成的治疗性调节。在……里面 在这个修订后的项目中，我们寻求开发抗体和多肽来调节 血管内皮细胞(EC)血管抑素受体的血管生成 我们发现它是细胞表面的三磷酸腺苷合成酶。细胞表面的三磷酸腺苷合成酶 低氧条件下肿瘤EC可能在补充能量产生中发挥作用 氧条件或内皮细胞信号事件与肿瘤的关系 血管生成。抗三磷酸腺苷合成酶亚基多克隆抗体竞争 用血管抑素进行细胞表面EC的结合和阻断能力 血管抑素抑制EC的增殖和迁移。多克隆抗血清 对b-亚基的抑制也表现出超过 血管抑素。此外，我们最近发现了另一种产生ATP的 EC表面的酶，核苷二磷酸激酶H1(NDPK H1)，即 被血管抑素显著抑制(提交发表的手稿， 见附录)。这些和其他发现支持血管抑素的假设 通过破坏血管内皮细胞表面三磷酸腺苷发挥抗增殖作用 综合。很可能是公认的血管抑素抑制能力 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 治疗药物在人类中的应用，因为生产、稳定性、 和亲和力。我们发现血管抑素的两个EC靶点提供了 开发更强健的化合物以阻断血管生成的可能性 人类癌症和其他增殖性疾病。这个项目的目标是 开发可替代血管抑素的抗体和多肽以抑制 乳腺癌中与三磷酸腺苷直接相互作用的血管生成 EC表面的合成装置。这些化合物有望成为 治疗剂证明了密集努力开发适当的 人源化单抗和靶向多肽噬菌体展示文库。 将筛选候选抗体和多肽以检测EC抑制活性 使用已建立的检测增殖、迁移和输卵管的方法 队形。此外，我们还开发了一种新的测定EC表面ATP的方法 将用于筛选化合物的抑制活性的合成。 在任何一种检测中表现出抑制活性的化合物将是 利用人的乳房筛选它们抑制体内肿瘤生长的能力 肿瘤异种移植模型。由于暴露的血管内定位 ATP合成酶，这个项目将产生一系列 抗血管生成化合物具有很强的快速转化为 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.