Vascular regulation by kringles of HGF and molecular cloning of a novel kringle-containing protein

HGF 的三环对血管的调节以及一种新型三环蛋白的分子克隆

基本信息

批准号：
11680631
负责人：
MATSUMOTO Kunio
金额：
$ 2.37万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11680631/
关键词：
HGF kringle domain NK4 angiogenesis tumor angiogenesis Kremen metastasis pancreatic cancer 肝細胞増殖因子 HGT 癌治療クリングル制癌剤

项目摘要

Various proteins containing kringle domains play diverse roles in tissue regeneration, blood coagulation/fibrinolysis, neural development, and blood vessel regulation. Identification of a novel kringle-containing molecule is likely to lead to further understanding of blood vessel regulation, including angiogenesis and anti-angiogenesis. In the present study, to study regulatory mechanisms by which kringle domains exhibit angio-regulatory functions, we analyzed angiostatic action of NK4, previously identified as HGF-antagonist. Likewise, we succeeded in molecular cloning of a novel kringle-containg molecule, Kremen.1) NK4 as angiogenesis inhibitor : NK4, a four kringles-cpontaining HGF-antagonist, inhibited proliferation and migration of vascular endothelial cells in culture. Likewise, NK4 inhibited angiogenesis in CAM and cornea assays in vivo. Together with a previously characterized biological action of NK4 as HGF-antagonist, therapeutic potential of NK4 for treatment of metastatic c … More ancers were tested. NK4 administered into mice potently suppressed metastasis, tumor angiogenesis, and tumor growth of cancers, including lung, breast, and pancreatic cancers. NK4 will become a potent anti-cancer agent, targeting malignant behavior of cancer.2) Using a newly established cloning strategy for kringle-containg proteins, 'Kringle-SAGE', we cloned cDNA for Kremen. Kremen is composed of 473 amino acids and is type-I transmembrane protein. Kremen contains unique domains, kringle domain and CUB domain in its extracellular region, while it has no specific conserved functional motif in its intracellular region. Kremen mRNA is expressed in diverse tissues, while it is up-regulated during muscular and neural differentiation. Kringle domain of Although Kremen may have some roles in vascular regulation and differentiation of muscular and neural cells, its roles remain to be further addressed. A newly established method for comprehensive cloning of kringle-proteins, Kringle-SAGE is powerful in molecular cloning of kringle proteins. Less

含有 kringle 结构域的各种蛋白质在组织再生、血液凝固/纤维蛋白溶解、神经发育和血管调节中发挥着不同的作用，识别一种新型的含有 kringle 的分子可能会导致对血管调节的进一步了解，包括血管生成和抗血管生成。在本研究中，为了研究 kringle 结构域表现出血管调节功能的调节机制，我们分析了 NK4（之前被鉴定为 HGF 拮抗剂）的血管抑制作用。成功地分子克隆了一种新的含三环分子，Kremen。1) NK4 作为血管生成抑制剂：NK4 是一种四三环 HGF 拮抗剂，可抑制培养物中血管内皮细胞的增殖和迁移。类似地，NK4 也可抑制 CAM 中的血管生成。和体内角膜测定连同先前表征的 NK4 作为 HGF 拮抗剂的生物作用，NK4 的治疗潜力。对转移性癌症进行了测试，NK4 可有效抑制癌症（包括肺癌、乳腺癌和胰腺癌）的转移、肿瘤血管生成和肿瘤生长。 .2) 使用新建立的含 kringle 蛋白的克隆策略“Kringle-SAGE”，我们克隆了 Kremen 的 cDNA，其组成为： Kremen 蛋白由 473 个氨基酸组成，是一种 I 型跨膜蛋白，其胞外区含有独特的结构域、kringle 结构域和 CUB 结构域，而其胞内区没有特定的保守功能基序。尽管 Kremen 可能在肌肉和神经细胞的血管调节和分化中发挥一定作用，但其作用仍有待进一步研究。 kringle-蛋白质，Kringle-SAGE 在 kringle 蛋白质的分子克隆方面功能强大。