Biological characterization of matched primary colorectal carcinomas & metastases

匹配的原发性结直肠癌的生物学特征

• 批准号: 8635138
• 负责人: Efsevia Vakiani
• 金额: $ 17.87万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-18 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8635138
• 关键词: Accounting; Address; Affect; Animal Model; Area; BRAF gene; Biological; Biological Assay; Biological Models; Cancer Biology; Cancer Etiology; Cancer Patient; Cell Line; Cessation of life; Clonal Evolution; Collection; Colorectal Cancer; Colorectal Neoplasms; Computer Analysis; Custom; DNA; Development; Disease; Disease Progression; Epigenetic Process; Event; Formalin; Fostering; Freezing; Gene Frequency; Gene Mutation; Gene Silencing; Genes; Genetic; Genetic Heterogeneity; Genomics; Goals; Health; Heterogeneity; Homing; In Vitro; Injection of therapeutic agent; Institution; KRAS2 gene; Laboratories; Large Intestine Carcinoma; MALDI-TOF Mass Spectrometry; Malignant Epithelial Cell; Malignant Neoplasms; Memorial Sloan-Kettering Cancer Center; Mentors; Mentorship; Metastatic Lesion; Metastatic to; Methylation; Mutate; Mutation; NRAS gene; Neoplasm Metastasis; PIK3CA gene; Paraffin Embedding; Pathology; Pattern; Point Mutation; Primary Lesion; Primary Neoplasm; Principal Investigator; Process; Recording of previous events; Recurrence; Relative (related person); Research; Research Project Grants; Role; Sampling; Scientist; Sensitivity and Specificity; Site; Surgical Oncology; TP53 gene; Tail; Tissues; Training; Tumor Suppressor Genes; Tumor Suppressor Proteins; United States; Veins; Work; adenoma; anticancer research; base; career development; cell motility; cohort; design; epigenomics; experience; gastrointestinal; genome analysis; in vivo; insertion/deletion mutation; insight; knowledge base; member; metastatic colorectal; mortality; next generation sequencing; novel; novel therapeutics; oncology service; programs; promoter; protein expression; skills; technology development; therapeutic target; tumor

DESCRIPTION (provided by applicant): This proposal describes a 5 year mentored research project designed to transition the applicant into an independent translational scientist. During th project period, the Principal Investigator (P.I.) will devote efforts towards broadening her knowledge base and technical skills as necessary to ultimately supervise a successful laboratory based program in cancer biology. The development of the P.I. will be fostered by ongoing mentorship by an established expert in the field of cancer biology, Dr. David Solit, at an institution renowned for training clinician scientists in cancer research, Memorial Sloan-Kettering Cancer Center. To further assist with her research project and career development, the P.I. and her primary mentor have assembled a multi-disciplinary mentorship team composed of Drs. Michael Berger and William Lee, experts in genomic technology development and computational analysis, Dr. Timothy Chan, an expert in epigenomics, Dr. Leonard Saltz, Chief of the Gastrointestinal Oncology Service, Martin Weiser, Associate Member of the GI Surgical Oncology Service, and David Klimstra, Chair of the Department of Pathology. The focus of the applicant's research is the biological characterization of matched primary colorectal carcinomas (CRC) and metastases. In Aim 1, we will use a novel bait-capture, next generation sequencing assay to define the mutational status of 300 cancer associated genes in 100 primary CRC and matched metastases. Matched pairs that show discordant mutational profiles between the primary tumor and the metastasis will be further analyzed using the same bait-capture, next generation sequencing assay, to sequence multiple areas of the primary tumor and metastasis in order to determine whether discordance when present can be attributed to intratumoral genetic heterogeneity. Whole genome analysis will be performed in a select subset of cases. In Aim 2, we will focus on tumor suppressor genes that are found in Aim 1 to be more commonly mutated in the metastatic lesions versus their corresponding primary tumors. These genes will be studied for differences in methylation patterns and protein expression levels. In Aim 3, we will study discordant alterations identified in Aim 1 to determine whether they have a functional role in invasion or metastasis. In these functional studies, we will use isogenic CRC cell lines to assess the effects of gene mutations on cell migration and invasion. Genes will then be further studied in vivo using a tail vein injection tumor metastasis model system. The overall goal of these studies is to define the extent of genetic and epigenetic heterogeneity in metastatic CRC and to identify genetic alterations that drive disease progression in colorectal cancer patients, thus, aiding in the identification of novel therapeutic strategies for this disease.

描述（由申请人提供）：该提案描述了一个为期 5 年的指导研究项目，旨在将申请人转变为独立的转化科学家。在项目期间，首席研究员（P.I.）将致力于扩大她的知识基础和必要的技术技能，以最终监督癌症生物学实验室项目的成功。 P.I. 的发展该项目将得到癌症生物学领域知名专家 David Solit 博士的持续指导，该机构以培训癌症研究临床科学家而闻名，纪念斯隆-凯特琳癌症中心 癌症中心。为了进一步协助她的研究项目和职业发展，P.I.和她的主要导师组建了一个由博士组成的多学科指导团队。基因组技术开发和计算分析专家 Michael Berger 和 William Lee、表观基因组学专家 Timothy Chan 博士、胃肠肿瘤服务主任 Leonard Saltz 博士、胃肠道肿瘤服务副会员 Martin Weiser 以及大卫·克利姆斯特拉 (David Klimstra)，病理学系主任。申请人的研究重点是匹配的原发性结直肠癌（CRC）和转移瘤的生物学特征。在目标 1 中，我们将使用新型诱饵捕获、下一代测序测定来确定 100 个原发性 CRC 和匹配的转移灶中 300 个癌症相关基因的突变状态。将使用相同的诱饵捕获、下一代测序测定进一步分析显示原发肿瘤和转移瘤之间不一致突变谱的匹配对，以对原发肿瘤和转移瘤的多个区域进行测序，以确定是否可以排除存在的不一致。归因于瘤内遗传异质性。全基因组分析将在选定的病例子集中进行。在目标 2 中，我们将重点关注目标 1 中发现的肿瘤抑制基因，这些基因在转移性病变中比相应的原发性肿瘤中更常见突变。将研究这些基因的甲基化模式和蛋白质表达水平的差异。在目标 3 中，我们将 研究目标 1 中发现的不一致改变，以确定它们是否在侵袭或转移中具有功能性作用。在这些功能研究中，我们将使用同基因CRC细胞系来评估基因突变对细胞迁移和侵袭的影响。然后将使用尾静脉注射肿瘤转移模型系统在体内进一步研究基因。这些研究的总体目标是确定转移性结直肠癌遗传和表观遗传异质性的程度，并确定驱动结直肠癌患者疾病进展的遗传改变，从而帮助确定该疾病的新治疗策略。