THE GENETICS OF OCULAR MELANOMA

眼部黑色素瘤的遗传学

基本信息

批准号：
8179029
负责人：
Anne Mary Bowcock
金额：
$ 59.77万
依托单位：
WASHINGTON UNIVERSITY
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 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8179029
关键词：
Automobile Driving BARD1 gene BRCA1 Associated Protein-1 BRCA1 gene BRCA2 gene Binding Binding Proteins Biological Assay Blood Cancer Etiology Cell Line Cellular Morphology Cessation of life Characteristics Chromosomes, Human, Pair 3 Classification Clinical Code Cytogenetics DNA DNA Sequence Development Diagnosis Diagnostic Dideoxy Chain Termination DNA Sequencing Disease Eye Neoplasms Frequencies Gene Expression Gene Expression Profile Gene Mutation Genes Genetic Genomic Instability Genomics Goals Lead Liver Malignant Neoplasms Maps 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 Screening procedure 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 expectation follow-up high risk insight malignant breast neoplasm malignant neoplasm of eye melanoma multidisciplinary novel personal narratives prognostic success tumor tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): 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. ) PUBLIC HEALTH RELEVANCE: Personal Narrative. Uveal (ocular melanoma) is the most common primary cancer of the eye and the second most common form of melanoma. It can be classified according to their risk of death due to metastasis. Unlike some cancers, UM is relatively simple, and there are likely to be a tractable number of genetic alterations driving their development. We recently used the approach of exome capture and NexGen sequencing to identify a metastasis suppressor for UM. The goal of the current application is to identify the remaining important cancer genes. This has both diagnostic and prognostic implications, and may lead to the development of novel forms of treatment for this devastating cancer.

描述（由申请人提供）：摘要：转移是恶性肿瘤的一个明显特征，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部癌症，也是第二常见的黑色素瘤形式。 UM 具有高度特征性的肝脏转移模式，对传统化疗具有抵抗力，并且通常是致命的。 UMs 的基因组不稳定性非常小，细胞遗传学改变很少，基因突变也很少见。因此，当在这些肿瘤中发现突变时，它们很可能是驱动突变而不是乘客突变。根据经过验证的基因表达特征，UM 可根据转移性死亡风险分为 1 类（低风险）和 2 类（高风险）。研究的一个主要焦点是确定赋予 UM 转移能力的特定基因变化。 2 类特征通常伴随着 3 号染色体的一份拷贝（单体 3）的丢失，这导致人们普遍认为，UM 细胞中 3 号染色体的一份拷贝的丢失会揭示 UM 细胞中一个或多个基因的隐性失活突变。赋予转移能力的 3 号染色体的剩余副本。其他变化包括 8q 染色体的增加和 8p 染色体的损失。在 1 类肿瘤中，6p 染色体的增加很常见。该提案的研究人员是密西根大学分子遗传学分析的先驱。我们是第一组应用最近描述的外显子组捕获技术，然后进行大规模并行测序，以将 BAP1 识别为 UM 中染色体 3 的转移抑制因子。在当前的研究中，我们将利用这些技术最近取得的成功来识别 UM 中的其他肿瘤抑制基因和癌基因突变。在目标 1 中，我们将生成 1 类和 2 类肿瘤的额外外显子组序列，将它们与匹配的种系 DNA 进行比较。含有有害突变的潜在抑癌基因和含有驱动 UM 发展的潜在激活突变的癌基因将通过 Sanger 测序进行确认，并在另外 10-30 个 1 类和 2 类肿瘤以及匹配的种系 DNA 中进行评估。在目标 2 中，我们将使用靶向捕获对其他样本（每种肿瘤类型超过 100 个）中这些基因内新发现的其他突变进行重新测序，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，以评估错义和框内编码变化对与已知和新伙伴相互作用的影响，并过度表达激活癌基因和敲低肿瘤抑制基因，以确定它们对细胞形态和基因表达的影响。然后，肿瘤分子改变的信息将与临床信息相结合，开始制定 UM 的预后分类。这是来自眼科和遗传学部门的多学科协作小组的协作提案，该小组拥有经过验证的专业知识来完成该提案的目标。）公共卫生相关性：个人叙述。葡萄膜（眼部黑色素瘤）是最常见的原发性眼部癌症，也是第二常见的黑色素瘤形式。它可以根据因转移而死亡的风险进行分类。与某些癌症不同，UM 相对简单，并且可能存在大量易于处理的基因改变来驱动其发展。我们最近使用外显子组捕获和 NexGen 测序的方法来鉴定 UM 的转移抑制因子。当前应用的目标是识别剩余的重要癌症基因。这具有诊断和预后意义，并可能导致针对这种毁灭性癌症的新型治疗方法的开发。