THE GENETICS OF OCULAR MELANOMA

眼部黑色素瘤的遗传学

基本信息

批准号：
8826698
负责人：
Anne Mary Bowcock
金额：
$ 40.38万
依托单位：
IMPERIAL COLLEGE OF SCIENCE, TECHNOLOGY AND MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8826698
关键词：
Automobile Driving BARD1 gene BRCA1 Associated Protein-1 BRCA1 gene BRCA2 gene Binding Binding Proteins Biological Assay Blood Breast Cancer gene Cancer Etiology Cell Line Cellular Morphology Cessation of life Characteristics Chromosomes, Human, Pair 3 Classification Clinical Code Cytogenetics DNA DNA Sequence DNA Sequence Alteration Development Diagnosis Diagnostic Dideoxy Chain Termination DNA Sequencing Disease Eye Neoplasms Frequencies Gene Expression Gene Expression Profile Genes Genetic Genomic Instability Genomic Segment Genomic approach Goals Lead Liver Malignant Neoplasms Maps Massive Parallel Sequencing Melanoma Cell Metastatic Neoplasm to the Liver Methods Molecular Molecular Genetics Monosomy Mutate Mutation Neoplasm Metastasis Ocular Melanoma Oncogenes Oncogenic Oncologist Ophthalmology Outpatients Pathway interactions Patients Pattern Protein Isoforms Research Research Personnel Resistance Retinoblastoma Risk Role Sampling Science Somatic Mutation Susceptibility Gene Techniques Technology Tertiary Protein Structure Tumor Suppressor Genes Tumor Suppressor Proteins Uveal Melanoma Work abstracting base cancer risk chemotherapy chromosome 6p gain chromosome 8p loss chromosome 8q gain early onset exome exome sequencing expectation follow-up high risk insight malignant neoplasm of eye melanoma multidisciplinary novel personal narratives prognostic screening success tumor tumorigenesis

项目摘要

Abstract: Metastasis is a defining feature of malignant tumors and is the most common cause of cancer-related death. However, the genetics of metastasis are poorly understood. Uveal melanoma (UM) is the most common primary cancer of the eye and the second most common form of melanoma. UMs have a highly characteristic pattern of metastasis to the liver that is resistant to conventional chemotherapy and is usually fatal. UMs have remarkably little genomic instability, few cytogenetic alterations, and rare genetic mutations. Thus, when mutations are found in these tumors, they are highly likely to be driver rather than passenger mutations. UMs can be grouped according to risk of metastatic death into class 1 (low risk) and class 2 (high risk) based on a validated gene expression signature. A major focus of research has been to identify the specific genetic changes that confer metastatic competency in UM. The class 2 signature is usually accompanied by loss of one copy of chromosome 3 (monosomy 3), which has led to the widespread expectation that loss of one copy of chromosome 3 in UM cells unmasks recessive inactivating mutations in a gene (or genes) on the remaining copy of chromosome 3 that confer metastatic capacity. Other changes include gain of chromosome 8q and loss of chromosome 8p. In the class of class 1 tumors, gain of chromosome 6p is common. The)investigators of this proposal are pioneers in the analysis of the molecular genetics of UM. We were the first group to apply the recently described technique of exome capture followed by massively parallel sequencing to identify BAP1 as the metastasis suppressor mapping to chromosome 3 in UM. In the current study we will capitalize on our recent success with these technologies to identify additional tumor suppressor genes and oncogenes mutated in UM. In Aim 1 we will generate additional exome sequences of both class 1 and class 2 tumors, comparing them with their matched germline DNA. Potential tumor suppressor genes harboring deleterious mutations, and oncogenes harboring potential activating mutations driving the development of UM will be confirmed with Sanger sequencing and evaluated in an additional 10-30 class 1 and class 2 tumors and matched germline DNA. In Aim 2 we will use targeted capture to re-sequence newly identified additional mutations within these genes in additional samples (>100 of each tumor type) to determine their contribution to UM. In Aim 3 we will perform limited functional studies of 5 newly identified genes in cell lines. We will perform binding assays to evaluate the effects of mis-sense and in-frame coding changes on interactions with known and novel partners, and over-express activating oncogenes and knockdown tumor suppressors to determine their effect on cell morphology and gene expression. Information on molecular alterations in tumors will then be incorporated with clinical information to begin to develop a prognostic classification of UM. This is a collaborative proposal from a multidisciplinary collaborative group from the departments of Ophthalmology and Genetics that has the proven expertise to complete the aims of this proposal. )

抽象的：转移是恶性肿瘤的一个决定性特征，也是癌症相关死亡的最常见原因。然而，人们对转移的遗传学知之甚少。葡萄膜黑色素瘤 (UM) 是最常见的原发性眼癌和第二常见的黑色素瘤。 UM 具有很强的特征对常规化疗有抵抗力且通常致命的肝脏转移模式。 UM 有基因组不稳定性极低，细胞遗传学改变极少，基因突变罕见。因此，当在这些肿瘤中发现了突变，它们很可能是驱动突变而不是乘客突变。 UM 根据转移性死亡风险可分为 1 类（低风险）和 2 类（高风险）经验证的基因表达特征。研究的一个主要重点是确定特定的基因赋予 UM 转移能力的变化。 2 类签名通常伴随着丢失 3 号染色体的一份拷贝（单体 3），这导致人们普遍认为丢失一份拷贝 UM 细胞中 3 号染色体的染色体揭示了其余基因上一个或多个基因的隐性失活突变赋予转移能力的 3 号染色体副本。其他变化包括染色体 8q 的增加和 8p 染色体缺失。在 1 类肿瘤中，6p 染色体的增加很常见。该）调查员这项提议的提出者是密西根大学分子遗传学分析的先驱。我们是第一批申请的最近描述的外显子组捕获技术，然后进行大规模并行测序来识别 BAP1 作为 UM 中 3 号染色体的转移抑制因子。在当前的研究中，我们将利用我们的最近利用这些技术成功地识别了其他肿瘤抑制基因和致癌基因突变在密歇根大学。在目标 1 中，我们将生成 1 类和 2 类肿瘤的额外外显子序列，比较它们具有匹配的种系 DNA。潜在的肿瘤抑制基因含有有害突变，具有驱动 UM 发展的潜在激活突变的癌基因将得到证实对另外 10-30 个 1 类和 2 类肿瘤以及匹配的种系进行 Sanger 测序和评估脱氧核糖核酸。在目标 2 中，我们将使用靶向捕获对这些新发现的其他突变进行重新测序。额外样本（每种肿瘤类型超过 100 个）中的基因，以确定它们对 UM 的贡献。在目标 3 中，我们将对细胞系中 5 个新发现的基因进行有限的功能研究。我们将进行结合测定评估误判和框架内编码变化对与已知和新伙伴的交互的影响，并过度表达激活癌基因和敲低肿瘤抑制基因，以确定它们对细胞的影响形态和基因表达。有关肿瘤分子改变的信息将被纳入临床信息开始制定 UM 的预后分类。这是来自一个合作提案来自眼科和遗传学部门的多学科协作小组，拥有经过验证的专业知识来完成本提案的目标。）