Inhibition of Migration by ATE+31

ATE 31 抑制迁移

• 批准号: 7111158
• 负责人: EUGENE G LEVIN
• 金额: $ 39.28万
• 依托单位: TORREY PINES INST FOR MOLECULAR STUDIES
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-12-15 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7111158
• 关键词: MCF7 cell; RNA interference; angiogenesis; cell migration; enzyme activity; estrogen receptors; fibroblast growth factor; focal adhesion kinase; green fluorescent proteins; growth factor receptors; immunocytochemistry; immunoprecipitation; laboratory mouse; neoplastic growth; phosphorylation; protein structure function; receptor binding; synthetic peptide; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): The high molecular weight form of FGF-2 (24kDa FGF-2) is produced by alternative translational start sites and consists of the more commonly studied 18kDa FGF-2 with an additional 55 amino acids at the amino terminal end. Treatment of endothelial or breast carcinoma cells with 24kDa FGF-2 results in an 80% decrease in migration rate although it continues to promote cell proliferation. We have performed deletion mutagenesis and developed a truncated form of 24kD FGF-2 which is highly effective in inhibiting migration in vitro while having no effect on cell growth. This peptide consists of the amino terminal 86 amino acids of 24kD FGF-2 and is referred to as ATE+31. Studies in animals have shown that ATE+31 is highly effective in suppressing mammary carcinoma and lung carcinoma tumor growth by two mechanisms, suppressing the angiogenic response and inhibiting tumor cell migration away from the tumor core. Studies of the mechanism by which ATE+31 inhibits migration have revealed that growth factor-induced increases in focal adhesion kinase (FAK) phosphorylation are abrogated by ATE+31. The decrease in phosphorylation occurred at tyr397 ,tyr407, and tyr861. Concomitantly, exposure to ATE+31 inhibited the reorganization of micro filaments and loss of focal adhesions that accompany growth factor treatment making enhanced motility difficult to attain. The hypothesis that this project will address states that ATE+31 inhibits cell migration through a mechanism involving the suppression of FAK function through inhibition of enhanced phosphorylation. The project will examine the mechanism by which ATE-31 inhibits cell migration and employ animal models to test the in vitro results in vivo. Specifically the project will define the mechanisms by which ATE+31 suppresses FAK phosphorylation and how it subsequently inhibits focal adhesion/rnicrofi lament dynamics and employ mouse models of angiogenesis and tumor development to test the experimental results in vivo. In addition, the therapeutic potential of ATE+31 will be evaluated in animal models of tumor development, regression, and metastasis will be evaluated.

描述（由申请人提供）：FGF-2的高分子量形式（24 kDa FGF-2）由替代翻译起始位点产生，由更常研究的18 kDa FGF-2和氨基末端的额外55个氨基酸组成。用24 kDa FGF-2处理内皮细胞或乳腺癌细胞导致迁移率降低80%，尽管它继续促进细胞增殖。我们已经进行了缺失突变，并开发了一种截短形式的24 kD FGF-2，这是非常有效的抑制迁移在体外，而对细胞生长没有影响。该肽由24 kD FGF-2的氨基末端86个氨基酸组成，称为ATE+31。动物研究表明，ATE+31通过两种机制，即抑制血管生成反应和抑制肿瘤细胞迁移远离肿瘤核心，在抑制乳腺癌和肺癌肿瘤生长方面非常有效。ATE+31抑制迁移的机制研究表明，生长因子诱导的粘着斑激酶（FAK）磷酸化的增加被ATE+31消除。磷酸化的减少发生在tyr 397、tyr 407和tyr 861处。同时，暴露于ATE+31抑制了伴随生长因子处理的微丝重组和局灶性粘连丧失，使得难以获得增强的运动性。该项目将解决的假设指出，ATE+31通过一种机制抑制细胞迁移，该机制涉及通过抑制增强的磷酸化抑制FAK功能。该项目将研究ATE-31抑制细胞迁移的机制，并采用动物模型来测试体内的体外结果。具体而言，该项目将定义ATE+31抑制FAK磷酸化的机制以及它随后如何抑制粘着斑/微纤维动态，并采用血管生成和肿瘤发展的小鼠模型来测试体内实验结果。此外，还将在肿瘤发展、消退和转移的动物模型中评价ATE+31的治疗潜力。