Isogenic Cell-Based Screen:Cancer-Targeting Catenin(RMI)

基于同基因细胞的筛选：癌症靶向连环蛋白 (RMI)

• 批准号: 7022520
• 负责人: TODD A WALDMAN
• 金额: $ 7.76万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7022520
• 关键词: antineoplastics; biotechnology; cadherins; cell line; colon neoplasms; cytotoxicity; drug screening /evaluation; gene mutation; high throughput technology; neoplastic cell; oncogenes; small molecule; technology /technique development

DESCRIPTION (provided by applicant): Activating mutations in the B-catenin oncogene are found in a wide range of common human cancers, including colorectal carcinoma, hepatocellular carcinoma, endometrial carcinoma, thyroid carcinoma, and others. Activitated oncogenes such as B-catenin are an appealing cancer drug target, since they represent a required genetic gain-of-function found in cancer cells but not in the patient's normal tissues. Consistent with this idea, we have previously experimentally demonstrated that targeting oncogenic B-catenin is likely to be an effective anticancer strategy (Mol Can Ther 1:1355, 2002). In this application we propose to implement a cell-based screen to identify small molecule and natural product lead compounds that are specifically cytotoxic/cytostatic to human colon cancer cells harboring an allele of oncogenic B-catenin. To do this we will employ a paired set of gene-targeted human cancer cells that differ only in the presence or absence of their endogenous allele of mutant (oncogenic) B-catenin. Since the cells are otherwise isogenic, compounds that specifically kill or inhibit the growth of parental cells (harboring oncogenic B-catenin) but not the derivatives in which oncogenic B-catenin has been deleted by gene targeting are good candidates as B-catenin targeted lead compounds. Such hits will be further validated in a variety of secondary screens to demonstrate reproducibility and generalizability. Once feasibility has been demonstrated by screening approximately 4000 compounds, we will apply to transfer the screen to an NIH Roadmap screening center for high throughput screening efforts.

描述（由申请人提供）：在广泛的人类癌症中发现了B-catenin癌基因中的激活突变，包括结直肠癌，肝细胞癌，子宫内膜癌，甲状腺癌等。激活的癌基因（例如B-钙蛋白）是一个有吸引力的癌症药物靶标，因为它们代表了癌细胞中所需的遗传功能，但在患者的正常组织中却没有。与这个想法一致，我们以前已经在实验上证明，靶向致癌性B-catenin可能是一种有效的抗癌策略（Mol可以1：1355，2002）。在此应用中，我们建议实施一个基于细胞的筛选，以鉴定针对具有致癌性B-catenin等位基因的人类结肠癌细胞，这些分子和天然产物铅化合物特异性细胞毒性/细胞抑制剂。为此，我们将采用一套成对的基因靶向人类癌细胞，仅在存在或不存在突变体（致癌）B-catenin的内源性等位基因的情况下有所不同。由于细胞否则是同种源的，因此特异性杀死或抑制父母细胞的生长（含有致癌性B-catenin）的化合物，而不是通过基因靶向删除致癌性B-catenin的衍生物，是B-catenin靶向铅铅铅铅的良好候选者化合物。此类命中将在各种二级筛选中得到进一步验证，以证明可重复性和概括性。一旦通过筛选大约4000种化合物来证明可行性，我们将申请将屏幕转移到NIH路线图筛查中心，以进行高吞吐量筛选工作。