Identification of the target molecules for antiangiogenic therapy

抗血管生成治疗靶分子的鉴定

• 批准号: 11671561
• 负责人: KANDA Shigeru
• 金额: $ 2.5万
• 依托单位: Nagasaki 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-11671561/
• 关键词: endothelial cells; angiogenesis; fibroblast growth factor receptor; anti-angiogenic therapy; c-Src; c-Fes; c-Fyn; tube formation; FGF; Src; Fes; 遊走; MAPキナーゼ

Head investigator has established a murine brain capillary endothelial cells (IBE cells) in 1996 and started the examination of FGF-2-mediated signal transduction pathways leading to angiogenesis. The purpose of the study is to identify specific signaling molecules involved in angiogenesis to make it possible to establish a new drug specifically and potently inhibit angiogenesis in vivo, such as tumor growth. IBE cells exhibit a panel of angiogenic cellular responses, such as urokinase production, proliferation, migration and tube formation. However, tube formation is rather specific for endothelial cells when compared with malignant tumor cells. By using this culture model, we found several important molecules as follows ;a. Blocking tube formation without any effect on proliferation by 4N1K peptide was enough to inhibit in vivo angiogenesis (mouse cornea micropocket assay). This result suggests the possibility that targeting tube formation is a good antiangiogenic strategy.b. c-Fes tyrosine kinase was involved in FGF-2-mediated chemotaxis and overexpression of wild type c-Fes induced FGF-2-independent tube formation.c. c-Src kinase was involved in chemotaxis toward FGF-2 by activating mitogen-activated protein kinase within focal adhesions.d. c-Fyn was transiently activated in IBE cells when cultured only on collagen gels (tube forming condition without proliferation or migration) and three stable cell lines expressing kinaseinactive c-Fyn exhibited attenuation of FGF-2-nediated tube formation (submitted). In the same study, it was found that c-Fes was activated by the adhesion of collagen gels. More recently, another growth factor was found to induce migration and tube formation. For migration, c-Fes was used and c-Fyn played a pivotal role in tube formation (submitted). Taken together, downstream molecules of c-Fes and c-Fyn would be potential targets of antiangiogenic therapy.

首席研究员于1996年建立了小鼠脑毛细血管内皮细胞（IBE细胞），并开始检查FGF-2介导的导致血管生成的信号转导途径。该研究的目的是鉴定参与血管生成的特异性信号分子，从而有可能建立特异性和有效抑制体内血管生成（如肿瘤生长）的新药。IBE细胞表现出一组血管生成细胞反应，如尿激酶产生、增殖、迁移和管形成。然而，与恶性肿瘤细胞相比，管形成对于内皮细胞是相当特异的。通过使用该培养模型，我们发现了如下几个重要分子：a.通过4 N1 K肽阻断管形成而对增殖没有任何影响足以抑制体内血管生成（小鼠角膜微袋测定）。该结果表明靶向管形成是一种良好的抗血管生成策略的可能性。c-Fes酪氨酸激酶参与FGF-2介导的趋化性和野生型c-Fes过表达诱导的FGF-2非依赖性小管形成。c-Src激酶通过激活粘着斑内的丝裂原活化蛋白激酶参与对FGF-2的趋化。当仅在胶原凝胶上培养时，c-Fyn在IBE细胞中被瞬时激活（管形成条件，没有增殖或迁移），并且三种表达激酶失活的c-Fyn的稳定细胞系表现出FGF-2介导的管形成的减弱（提交）。在同一研究中，发现c-Fes通过胶原凝胶的粘附而被激活。最近，发现另一种生长因子诱导迁移和管形成。对于迁移，使用c-Fes，c-Fyn在管形成中发挥关键作用（已提交）。综合考虑，c-Fes和c-Fyn的下游分子将成为抗血管生成治疗的潜在靶点。

项目成果

S.Kanda,et al.: "Fibroblast growth factor-mediated signal trausduction pathways involved in en angiogenic responses of endothelial cells"Current Topics in Biochemical Research. (印刷中). (2000)

S. Kanda 等人：“参与内皮细胞血管生成反应的成纤维细胞生长因子介导的信号传导途径”（Current Topics in Biochemical Research）（2000 年出版）。

T.Tsurusaki,S.Kanda,et al.: "Vascular endothelial growth factor-C expression in human prostatic carcinoma and its relationship to lymph node metastasis"British Journal of Cancer. 80. 309-313 (1999)

T.Tsurusaki，S.Kanda，等：“血管内皮生长因子-C在人前列腺癌中的表达及其与淋巴结转移的关系”英国癌症杂志。

Shigeru Kanda, Edwina C.Lerner, Satoshi Tsuda, Takefumi Shono, Hiroshi Kanetake, and Thomas E.Smithgall: "The non-receptor proteintyrosine kinase c-Fes is involved in FGF-2-induced chemotaxis of murine brain capillary endothelial cells."J.Biol.Chem.. 275.

Shigeru Kanda、Edwina C.Lerner、Satoshi Tsuda、Takefum​​i Shono、Hiroshi Kanetake 和 Thomas E.Smithgall：“非受体蛋白酪氨酸激酶 c-Fes 参与 FGF-2 诱导的小鼠脑毛细血管内皮细胞的趋化性。”

神田滋,宮園浩平: "内科学"黒川清,松澤祐次 編 文光堂. 3 (1999)

神田茂、宫园航平：《内科》黑川清、松泽佑二（编）文科堂 3（1999）。

Shigeru Kanda, Bianca Tomasini-Johansson, Kristofer Rubin and Lena Claesson-Welsh: "Signaling via fibroblast growth factor receptor-1 is dependent on extracellular matrix in capillary endothelial cell differentiation."Exp.Cell Res.. 248. 203-213 (1999)

Shigeru Kanda、Bianca Tomasini-Johansson、Kristofer Rubin 和 Lena Claesson-Welsh：“通过成纤维细胞生长因子受体 1 发出的信号依赖于毛细血管内皮细胞分化中的细胞外基质。”Exp.Cell Res.. 248. 203-213 (1999)