A Validated Resource of Thyroid Cancer Cell Lines for Pathway Discovery

用于途径发现的经过验证的甲状腺癌细胞系资源

• 批准号: 7842829
• 负责人: JAMES A FAGIN
• 金额: $ 50万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7842829
• 关键词: 1-Phosphatidylinositol 3-Kinase; AKT1 gene; Activation Analysis; Address; Advanced Malignant Neoplasm; Antineoplastic Agents; Area; BRAF gene; Back; Bioinformatics; CTNNB1 gene; Cancer Biology; Cancer Patient; Cancer cell line; Cell Line; Cell Survival; Cells; Collaborations; Collection; Colon Carcinoma; Communities; Comparative Study; Computational Biology; Consensus; Core Facility; Critical Pathways; DNA; DNA Fingerprinting; DNA copy number; Data; Databases; Derivation procedure; Development; Diagnosis; Disease; Disease Progression; Distant; Endocrine; Funding; Gene Expression; Generations; Genes; Genetic; Genome; Genomics; Genotype; Histologic; Human; Human Resources; In Vitro; Incidence; Institutes; Life; Link; Literature; Maintenance; Malignant Neoplasms; Malignant neoplasm of thyroid; Mass Spectrum Analysis; Memorial Sloan-Kettering Cancer Center; Metastatic Lesion; Methodology; Mitogen-Activated Protein Kinases; Molecular; Molecular Profiling; Mutation; National Cancer Institute; Neoplasm Metastasis; Occupations; Oncogenes; PIK3CA gene; Pathogenesis; Pathway interactions; Patients; Play; Preparation; Principal Investigator; Procedures; Protein Isoforms; Publications; Qualifying; Refractory; Reporting; Research; Resources; Role; Sampling; Screening procedure; Short Tandem Repeat; Signal Pathway; Signal Transduction; Solid; Solid Neoplasm; Staging; Suppressor Genes; Technology; The University of Colorado Cancer Center; Therapeutic; Thyroid Gland; Time; Tissues; Translational Research; Tumor Suppressor Genes; Tumor Tissue; United States; Wages; Western Blotting; advanced disease; anticancer research; base; cancer cell; cancer genome; effective therapy; experience; genetic profiling; in vivo; melanoma; mortality; new therapeutic target; novel; novel therapeutics; programs; public health relevance; quantum; repository; research study; response; success; symposium; therapeutic target; tool; tumor; working group

DESCRIPTION (provided by applicant): This application addresses the broad Challenge Area (15) Translational Science, 15-CA-103: Thyroid Cancer Cell Line Project which is in direct response to our recent discovery showing that 17 out of 40 "thyroid" cancer cell lines are either redundant or misidentified (i.e. of a different tumor lineage including melanoma and colon cancer). The remaining 23 lines are unique and are likely of thyroid origin based on the expression of a limited number of thyroid-specific genes. These cell lines, however, have not been genetically linked to tumors of origin or carefully characterized at a genetic or molecular level. This problem for the research community was reviewed at a recent NCI conference in which the Principal Investigators participated. The need for well- validated thyroid cancer cell lines to study mechanisms of cancer development and progression as well as discovery of novel therapeutic targets was emphasized. In this proposal, we will use three complementary strategies to provide a large panel of comprehensively characterized thyroid cancer cell lines representing primary and metastatic tumors of different histologic types and mutational subtypes (BRAF, RET/PTC, RAS, PIK3CA, AKT1, CTNNB1). We will focus on developing cell lines from poorly differentiated thyroid cancer as well as metastatic lesions that are more likely to harbor mutations that are not represented in the current panel of thyroid cancer cell lines. To establish new cell lines, we will use a parallel in vivo and in vitro approach to maximize our success rate. The new cell lines will be carefully characterized and compared to the corresponding tumor tissue of origin by short tandem repeat (STR) profiling to confirm their derivation. STR profiling and expression of thyroid-specific genes will be used investigate the origins of an additional 36 thyroid cell lines reported in the literature in order to assemble a complete panel of thyroid cancer cell lines. We will use expression arrays of authentic thyroid cancer cell lines to identify a signature that can be used to validate cell lines that have lost expression of thyroid-specific markers or from which patient DNA is unavailable. Finally, we will apply global molecular and genomic approaches with novel computational analyses to the new and existing thyroid cancer cell lines to uncover pathways important in thyroid cancer development and progression. All cell lines, as well as the genomic and gene expression data, will be made available to the research community through a central repository with the assistance of the NCI, and data will be accessible through the NCBI GEO database and UCSC genome browser. Feasibility and Impact: The generation of new cell lines from solid tumors is a challenging task. We propose to generate at least 10 new cell lines in a 12-15 month time period using parallel in vivo and in vitro approaches. There is significant expertise at the University of Colorado Cancer Center (UCCC) with a proven record of success in generating cell lines from solid cancers. We believe that this ambitious proposal is feasible within the two-year time frame because the unique collaboration between the UC Denver and MSKCC has all of these advanced technologies currently in place (new procedures to develop cell lines, gene expression platforms, mass spectrometry genotyping, CGH array analysis, mutational screen of key oncogenes and tumor suppressor genes and high throughput pathway activation analysis by Western blotting). A majority of the sample preparation and studies for the arrays and mutation screening will be performed by experienced personnel in well-equipped cores with rapid turn-around times. Furthermore, we are collaborating with nationally recognized bioinformatics and computational experts to apply novel computational biology strategies to identify important signaling pathways that will better define disease pathogenesis. Successful completion of these aims will provide approximately 40 well-characterized thyroid cancer cell lines for use by the research community to uncover mechanisms of thyroid cancer development, progression, metastatic potential and to serve as a platform for identification of novel therapeutic targets for advanced disease. Lay Summary: Thyroid cancer is the most common endocrine malignancy with an annual incidence of ~34,000. At least 300,000 people are living with a diagnosis of thyroid cancer in the United States, and more than 1,500 of them die each year from this disease. Permanent thyroid cancer cell lines derived from patient's cancers are critical to help us understand the causes of thyroid cancer, how the tumor grows and spreads, and how we can find new ways to treat advanced disease for which there is currently no cure. Recent studies by our research groups have shown that nearly half of the "thyroid" cancer cell lines in current use are not unique and many did not even originate from a thyroid cancer. In order to provide well characterized thyroid cancer cell lines for study by the research community, we will generate new thyroid cancer cell lines that can be linked back to the original tumor tissue. We will also develop approaches by which we can determine whether some of the existing thyroid cancer cell lines are authentic or not. We will develop genetic profiles of all the cancer cell lines, from which we will be able to characterize the abnormal signaling networks that may explain how these cancers develop. Once we know what makes these cancers grow, we will have a better understanding of how to treat them. With the help of the National Cancer Institute, we will make these cell lines widely available to the research community. PUBLIC HEALTH RELEVANCE: The generation of a well-characterized panel of thyroid cancer cell lines has significant relevance for public health as validated tools for the broader research community. Successful completion of the proposed aims will provide approximately 40 well-characterized thyroid cancer cell lines in a central repository for use by the research community to uncover mechanisms of thyroid cancer development, progression, metastatic potential and to serve as a platform for identification of novel therapeutic targets for many types of advanced cancer.

描述（由申请人提供）：本申请解决了广泛的挑战领域 (15) 转化科学，15-CA-103：甲状腺癌细胞系项目，该项目直接响应我们最近的发现，表明 40 种“甲状腺”癌中有 17 种细胞系要么是多余的，要么是错误识别的（即不同的肿瘤谱系，包括黑色素瘤和结肠癌）。其余 23 个品系是独特的，根据有限数量的甲状腺特异性基因的表达，很可能源自甲状腺。然而，这些细胞系尚未在遗传上与肿瘤起源相关，也没有在遗传或分子水平上进行仔细表征。最近由首席研究员参加的 NCI 会议回顾了研究界面临的这个问题。强调需要经过充分验证的甲状腺癌细胞系来研究癌症发生和进展的机制以及发现新的治疗靶点。在本提案中，我们将使用三种互补策略来提供一大批全面表征的甲状腺癌细胞系，代表不同组织学类型和突变亚型的原发性和转移性肿瘤（BRAF、RET/PTC、RAS、PIK3CA、AKT1、CTNNB1）。我们将重点开发低分化甲状腺癌和转移性病变的细胞系，这些细胞系更有可能含有当前甲状腺癌细胞系中未代表的突变。为了建立新的细胞系，我们将使用并行的体内和体外方法来最大限度地提高成功率。新细胞系将被仔细表征，并通过短串联重复（STR）分析与相应的肿瘤组织来源进行比较，以确认其来源。甲状腺特异性基因的 STR 分析和表达将用于研究文献中报道的另外 36 个甲状腺细胞系的起源，以便组装完整的甲状腺癌细胞系。我们将使用真实甲状腺癌细胞系的表达阵列来识别特征，该特征可用于验证甲状腺特异性标记物表达缺失或无法获取患者 DNA 的细胞系。最后，我们将应用全球分子和基因组方法以及新颖的计算分析来研究新的和现有的甲状腺癌细胞系，以揭示甲状腺癌发生和进展的重要途径。所有细胞系以及基因组和基因表达数据将在 NCI 的协助下通过中央存储库提供给研究界，并且可通过 NCBI GEO 数据库和 UCSC 基因组浏览器访问数据。可行性和影响：从实体瘤中产生新的细胞系是一项具有挑战性的任务。我们建议使用平行的体内和体外方法在 12-15 个月的时间内产生至少 10 个新细胞系。科罗拉多大学癌症中心 (UCCC) 拥有丰富的专业知识，在从实体癌生成细胞系方面拥有成功的记录。我们相信，这一雄心勃勃的提议在两年的时间内是可行的，因为加州大学丹佛分校和 MSKCC 之间的独特合作目前拥有所有这些先进技术（开发细胞系的新程序、基因表达平台、质谱基因分型、 CGH 阵列分析、关键癌基因和抑癌基因的突变筛选以及通过蛋白质印迹进行的高通量通路激活分析）。阵列和突变筛选的大部分样品制备和研究将由经验丰富的人员在设备精良的核心中进行，周转时间短。此外，我们正在与国家认可的生物信息学和计算专家合作，应用新颖的计算生物学策略来识别重要的信号通路，从而更好地定义疾病发病机制。成功完成这些目标将提供大约 40 个特征良好的甲状腺癌细胞系，供研究界使用，以揭示甲状腺癌发生、进展、转移潜力的机制，并作为识别晚期疾病新治疗靶点的平台。摘要：甲状腺癌是最常见的内分泌恶性肿瘤，年发病率约为 34,000 例。在美国，至少有 300,000 人被诊断患有甲状腺癌，每年有超过 1,500 人死于这种疾病。来自患者癌症的永久性甲状腺癌细胞系对于帮助我们了解甲状腺癌的原因、肿瘤如何生长和扩散以及我们如何找到治疗目前无法治愈的晚期疾病的新方法至关重要。我们的研究小组最近的研究表明，目前使用的近一半“甲状腺”癌细胞系并不是独特的，许多甚至不是源自甲状腺癌。为了提供特征明确的甲状腺癌细胞系供研究界研究，我们将产生可与原始肿瘤组织联系起来的新甲状腺癌细胞系。我们还将开发方法来确定一些现有的甲状腺癌细胞系是否真实。我们将开发所有癌细胞系的遗传图谱，从中我们将能够表征异常信号网络，从而解释这些癌症是如何发展的。一旦我们知道是什么导致这些癌症生长，我们就会更好地了解如何治疗它们。在国家癌症研究所的帮助下，我们将向研究界广泛提供这些细胞系。 公共健康相关性：一组特征明确的甲状腺癌细胞系的产生对于公共健康具有重要意义，作为更广泛研究界的有效工具。成功完成拟议目标将在中央存储库中提供大约 40 个特征良好的甲状腺癌细胞系，供研究界使用，以揭示甲状腺癌发生、进展、转移潜力的机制，并作为识别新疗法的平台许多类型的晚期癌症的靶标。