Harnessing Functional Genomics to Reveal Cancer Specific Determinants of Mitosis

利用功能基因组学揭示癌症有丝分裂的特异性决定因素

• 批准号: 7472660
• 负责人: Angelique Wright Whitehurst
• 金额: $ 9.44万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-21 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7472660
• 关键词: Algorithms; Apoptosis; Automobile Driving; Biological; Cell Survival; Cells; Chromosomal Instability; Classification; Collection; Defect; Dose; Effectiveness; Elements; Environment; Gametogenesis; Gene Expression; Gene Targeting; Genes; Genome; Growth; Human; Human Genome; Intervention; Libraries; Malignant Neoplasms; Malignant Spindle Cell Neoplasm; Malignant neoplasm of lung; Microtubules; Mitosis; Mitotic; Mitotic Spindle Apparatus; Mitotic spindle; Molecular; Neurons; Non-Small-Cell Lung Carcinoma; Normal Cell; Numbers; Paclitaxel; Phenotype; Population; Process; Proteins; Public Health; RNA Interference; Rate; Regulation; Screening procedure; Small Interfering RNA; Structure of parenchyma of lung; System; Therapeutic; Tumor Tissue; Work; Xenograft Model; Xenograft procedure; base; cancer cell; chemotherapy; defined contribution; design; functional genomics; in vivo; migration; molecular pathology; neoplastic cell; novel; prevent; programs; response; therapeutic target; tumor; tumor growth; tumorigenic

DESCRIPTION (provided by applicant): Widespread evidence suggests that a unifying principle governing the molecular pathology of cancer is the co-dependent aberrant regulation of core machinery driving proliferation and suppressing apoptosis. Anomalous proteins engaged to support a tumorigenic regulatory environment likely represent optimal intervention targets in a heterogenous population of cancer cells. The advent of RNAi-based functional genomics provides the opportunity to derive unbiased comprehensive collections of validated gene targets supporting critical biological systems outside the framework of preconceived notions of mechanistic relationships. Here, I have combined a high throughput cell-based one-well/one-gene screening platform with an arrayed genome-wide synthetic siRNA library for systematic interrogation of the molecular underpinnings of cancer cell chemoresponsiveness. NCI-H1155, a human non small cell lung cancer line, was employed for a paclitaxel-dependent synthetic lethal screen designed to identify gene targets that specifically reduce cell viability in the presence of otherwise suboptimal pactlitaxel concentrations. Using a stringent objective statistical algorithm to reduce false discovery rates below 5%, we isolated a panel of 87 genes that represent major fulcrums of the cancer cell autonomous response to abrogation of microtubule dynamics. Importantly, a number of these targets sensitize lung cancer cells to paclitaxel concentrations 1000-fold lower than otherwise required for a significant response, and reveal novel mechanistic relationships between cancer-associated aberrant gene expression programs and the basic cellular machinery required for robust mitotic progression. In the studies proposed herein, I plan to characterize the mechanism of action of these targets and determine if they are required for tumor cell survival and chemoresistance in vivo. My specific aims are to 1: Defining the contribution of novel paclitaxel sensitizers to cancer cell spindle assembly and function. 2: Validate effectiveness of candidate therapeutic targets with orthotopic xenograft models of lung cancer. 3: Isolate the core components of the cancer cell network that liberate chemoresistant tumors from mitotic catastrophe checkpoints. With respect to public health, these studies will aid in identifying both novel chemotherapeutics as well as novel combinations of existing chemotherapeutics to better treat cancer.

描述（由申请人提供）：广泛的证据表明，控制癌症分子病理学的统一原则是驱动增殖和抑制凋亡的核心机制的共同依赖性异常调节。参与支持致瘤调节环境的异常蛋白可能代表癌细胞异质群体中的最佳干预靶点。基于RNAi的功能基因组学的出现提供了在机械关系的先入为主的概念的框架之外获得支持关键生物系统的经验证的基因靶标的无偏见的全面集合的机会。在这里，我结合了一个高通量的基于细胞的单孔/单基因筛选平台与阵列全基因组合成siRNA文库的癌细胞化学反应性的分子基础的系统询问。NCI-H1155是一种人非小细胞肺癌细胞系，用于紫杉醇依赖性合成致死筛选，旨在鉴定在其他次优紫杉醇浓度存在下特异性降低细胞活力的基因靶标。使用严格的客观统计算法将错误发现率降低到5%以下，我们分离出一组87个基因，这些基因代表了癌细胞对微管动力学消除的自主反应的主要支点。重要的是，许多这些目标敏感肺癌细胞紫杉醇浓度低于1000倍，否则需要一个显着的反应，并揭示了癌症相关的异常基因表达程序和强大的有丝分裂进程所需的基本细胞机制之间的新的机制关系。在本文提出的研究中，我计划描述这些靶点的作用机制，并确定它们是否是体内肿瘤细胞存活和耐药性所必需的。我的具体目标是1：定义新型紫杉醇增敏剂对癌细胞纺锤体组装和功能的贡献。2：候选治疗靶点对肺癌原位异种移植模型的有效性。3：分离癌细胞网络的核心组成部分，将化疗耐药肿瘤从有丝分裂灾难检查点中解放出来。 关于公共卫生，这些研究将有助于确定新的化疗药物以及现有化疗药物的新组合，以更好地治疗癌症。