Modulation of Angiogenesis Via the Angiostatin Receptor

通过血管抑制素受体调节血管生成

• 批准号: 6475360
• 负责人: Salvatore V Pizzo
• 金额: $ 5.08万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6475360
• 关键词: SCID mouse; adenosine triphosphate; adenosinetriphosphatase; angiogenesis; angiogenesis inhibitors; angiostatins; breast neoplasms; cell migration; cell proliferation; chimeric proteins; enzyme activity; human tissue; monoclonal antibody; neoplasm /cancer immunotherapy; neutralizing antibody; nonhuman therapy evaluation; nucleoside diphosphate kinase; peptide library; receptor; vascular endothelium

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.

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”的响应。在 在这个修订后的项目中，我们寻求开发能够调节的抗体和肽 最近，通过内皮细胞（EC）血管抑制素受体进行血管生成 我们发现它是细胞表面 ATP 合酶。表面ATP合酶 肿瘤EC可能在低能量条件下补充能量产生中发挥作用 氧气条件或肿瘤涉及的内皮信号传导事件 血管生成。抗 ATP 合酶亚基竞争的多克隆抗体 与血管抑制素结合细胞表面 EC 并阻断 血管抑制素抑制EC增殖和迁移。多克隆抗血清 针对 b 亚基还表现出超过直接 EC 抑制作用 血管抑制素。此外，我们最近发现了第二种产生 ATP 的物质 EC表面的酶，核苷二磷酸激酶h1（NDPK h1），即 被血管抑制素显着抑制（提交出版的手稿， 见附录）。这些和其他发现支持血管抑制素的假设 通过破坏表面 ATP 对 EC 发挥抗增殖作用 合成。血管抑制素的公认能力很可能是抑制 许多体内肿瘤的生长和转移也是通过抑制 EC表面ATP合成。然而，血管抑制素本身作为药物的潜力较差。 由于生产、稳定性的限制，人类治疗剂 和亲和力。我们对血管抑制素的两个 EC 靶点的发现提供了 开发更强大的化合物来阻止血管生成的可能性 人类癌症和其他增殖性疾病。该项目的目标是 开发可以替代血管抑制素来抑制血管生成的抗体和肽 通过与 ATP 直接相互作用促进乳腺癌血管生成 EC表面合成装置。这些化合物的前景 治疗药物证明需要大力努力开发适当的治疗药物 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 使用已建立的测量增殖、迁移和管的测定方法 形成。此外，我们还开发了一种新的 EC 表面 ATP 检测方法 合成将用于筛选化合物的抑制活性。 在任何这些测定中表现出抑制活性的化合物将是 使用人类乳房筛选其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，该项目将产生一系列 具有快速转化为强大潜力的抗血管生成化合物 临床研究。