The Genetics of Ocular Melanoma

眼部黑色素瘤的遗传学

基本信息

批准号：
10343767
负责人：
Anne Mary Bowcock
金额：
$ 63.08万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
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 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10343767
关键词：
Address Age Alternative Splicing Antineoplastic Agents Applications Grants Automobile Driving BAP1 gene Biological Models Biological Sciences Blindness Cancer Model Candidate Disease Gene Cell Line Cessation of life Characteristics Chromatin Remodeling Factor Chromosome 3 Chromosome abnormality Chromosomes Clinical Competence Complement DNA Sequence Rearrangement DNA sequencing Detection Development Diagnosis Drosophila genus Epithelial Event Exhibits Eye Eye Neoplasms Future G-Protein-Coupled Receptors GNAQ gene Gene Expression Profile Gene Fusion Generations Genes Genetic Genome Genomic DNA Genomics Grant Human Immune checkpoint inhibitor Introns Lead Length Loss of Heterozygosity MEKs Malignant Neoplasms Maps Metastatic to Modeling Molecular Mutation Neoplasm Metastasis Nonmetastatic Ocular Melanoma Oncogenic Orthologous Gene Pathogenicity Pathway interactions Patients Phenotype Primary Neoplasm Prognosis Protein Isoforms RNA Recurrence Research Resolution Risk Signal Transduction Site Solid Neoplasm Somatic Mutation System Systemic Therapy Testing Tissues Transgenic Organisms Up-Regulation Uveal Melanoma Variant Wing Work cancer genome chromatin modification chromosome 8q gain chromosome loss cysteinyl leukotriene receptor 2 driver mutation effective therapy epigenomics exome experimental study fly guanine nucleotide binding protein high risk insertion/deletion mutation insight loss of function loss of function mutation malignant neoplasm of eye melanocyte melanoma monolayer novel therapeutics polypeptide single molecule success therapeutic candidate transcriptome sequencing tumor tumor diagnosis

项目摘要

PROJECT SUMMARY/ABSTRACT Uveal melanoma (UM) is the second most common form of melanoma and the most common primary cancer of the eye, resulting not only in vision loss, but in metastatic death in up to half of patients. There are no effective treatment options once the tumor metastasizes and patients usually die within a few months of diagnosis despite systemic therapy. UMs harbor activating oncogenic mutations in GNAQ, GNA11 or CYSLTR2 that occur early in tumor formation and that are unrelated to metastatic risk. We showed that over 80% of class 2 UMs harbor loss of function mutations of BAP1 and that recurrent mutations altering R625 of SF3B1 are found in tumors with slower rates of metastasis. Mutations in EIF1AX are associated with tumors with minimal chance of metastasis. The presence of BAP1, SF3B1 and EIF1AX mutations, are nearly always mutually exclusive. However, overlaid on this are genomic rearrangements that include gain of chromosomes 8q and 6p, and loss of chromosomes 1p, 6q and 8p. Our studies of these karyotypic/copy number alterations in UM have identified a critical chromatin modifier mapping to chromosome 6q (PHF10) and candidate genes in other regions of recurrent chromosomal alteration. The consequences of these changes in the development of UM are poorly understood. To extend and refine our copy number studies we will now will perform long-read single molecule sequencing of DNA and RNA isolated from primary UMs with different driver mutations and CNAs. In Aims 1 and 2 we will perform long-read single molecule sequencing (SMRT) to develop detailed maps of the structural variations (SVs) of the genomes of these tumors, identifying and characterizing critical breakpoints, gene fusions and tumor specific isoforms. Besides obtaining insights into the generation of chromosomal alterations we will validate and refine focal deletions and identify critical isoforms in such regions. Functional analyses in cell lines will examine the consequences of gene fusions and isoforms highly correlated with copy number changes. In parallel and in Aim 3 we will use the power of fly genetics to investigate the consequence of major alterations in UM in D. Melanogaster. We will first generate and characterize flies with loss of function mutations in the BAP1 ortholog calypso, and generate activating mutations of the GNAQ ortholog GαQ so that the combined effect of loss of calypso and activation of GαQ can be investigated. We will focus on larval eye discs and wing discs that represented well-ordered epithelial monolayers and examine the the phenotypic, signaling and epigenomic changes that arise from these mutations. The fly model is capable of addressing the functional relevance of the genomic insights from all three Aims by performing rapid genetic interactions. In the future, candidate UM drivers identified from Aims 1-2 will be tested for the ability to modify the cancer-like phenotypes we see in our two hit GαQ and calypso fly model. Genetic suppressors of overgrowth, tissue transformation, and metastasis phenotypes would represent exciting therapeutic candidates to pursue in future studies.

项目概要/摘要葡萄膜黑色素瘤 (UM) 是第二常见的黑色素瘤形式，也是最常见的原发性癌症眼睛，不仅导致视力丧失，而且导致多达一半的患者转移性死亡。目前尚无有效的治疗方法。一旦肿瘤转移，患者通常会在诊断后几个月内死亡，尽管 UMs 含有早期发生的 GNAQ、GNA11 或 CYSLTR2 激活致癌突变。我们发现超过 80% 的 2 类 UM 具有肿瘤形成的特征，并且与转移风险无关。 BAP1 功能缺失突变和改变 SF3B1 R625 的反复突变在患有以下疾病的肿瘤中发现： EIF1AX 突变与转移机会最小的肿瘤相关。然而，BAP1、SF3B1 和 EIF1AX 突变的存在几乎总是相互排斥的。这是基因组重排，包括获得 8q 和 6p 染色体，以及损失 1p 染色体，我们对 UM 中这些核型/拷贝数改变的研究已经确定了一个关键的染色质。映射到染色体 6q (PHF10) 的修饰符和复发染色体其他区域的候选基因人们对 UM 发展中这些变化的后果知之甚少。完善我们的拷贝数研究，我们现在将对 DNA 和 RNA 进行长读单分子测序从具有不同驱动突变和 CNA 的原代 UM 中分离出来。在目标 1 和 2 中，我们将进行长读。单分子测序（SMRT）以绘制基因组结构变异（SV）的详细图谱这些肿瘤的特征，识别和表征关键断点、基因融合和肿瘤特异性亚型。除了深入了解染色体改变的产生外，我们还将验证和完善焦点删除并识别这些区域中的关键亚型将检查细胞系中的功能。基因融合和亚型的后果与拷贝数变化高度相关。 3 我们将利用果蝇遗传学的力量来研究果蝇 UM 重大改变的后果。我们将首先生成并表征 BAP1 直系同源功能缺失突变的果蝇。 calypso，并产生 GNAQ 直系同源物 GαQ 的激活突变，从而使损失的综合效应我们将重点研究幼虫的眼盘和翼盘。代表有序的上皮单层并检查表型、信号传导和表观基因组这些突变引起的变化能够解决果蝇模型的功能相关性。通过执行快速的遗传相互作用，从所有三个目标中获得基因组见解。未来，候选 UM 驱动程序。将测试从目标 1-2 中确定的改变我们在两个命中中看到的癌症样表型的能力 GαQ 和 calypso 果蝇模型。过度生长、组织转化和转移的遗传抑制因子。表型将代表未来研究中令人兴奋的治疗候选者。