The Genetics of Ocular Melanoma

眼部黑色素瘤的遗传学

基本信息

批准号：
10596996
负责人：
Anne Mary Bowcock
金额：
$ 64.03万
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/10596996
关键词：
Abnormal Karyotype 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 Classification Clinical Competence Complement DNA Sequence Rearrangement DNA sequencing Detection Development Diagnosis Drosophila genus Epithelium Event Exhibits Eye Eye Neoplasms Future G-Protein-Coupled Receptors GNAQ gene GTP-Binding Proteins Gene Expression Profile Gene Fusion Generations Genes Genetic Genome Genomic DNA Genomics Grant Growth Human Immune checkpoint inhibitor Introns Length Loss of Heterozygosity MEKs Malignant Neoplasms Maps 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 1p loss chromosome 8q gain cysteinyl leukotriene receptor 2 driver mutation effective therapy epigenomics exome experimental study fly guanine nucleotide binding protein high risk 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.

项目摘要/摘要紫veal黑色素瘤（UM）是第二种最常见的黑色素瘤，也是最常见的原发性癌症眼睛不仅导致视力丧失，而且导致多达一半患者的转移性死亡。没有有效治疗选择一旦肿瘤转移，患者通常会在诊断目的地的几个月内死亡全身疗法。 UMS港口在早期发生的GNAQ，GNA11或CYSLTR2中激活致癌突变在肿瘤形成中，与转移风险无关。我们表明，超过80％的2级UMS港口在患有SF3B1的R625的BAP1的功能突变损失是在患有SF3B1的R625 转移速度较慢。 EIF1AX中的突变与转移机会最小的肿瘤有关。 BAP1，SF3B1和EIF1AX突变的存在几乎总是相互排斥的。但是，覆盖了因此是基因组重排，包括染色体8q和6p的增益以及染色体1p的损失， 6q和8p。我们对UM中这些核型/拷贝数改变的研究已经确定了关键的染色质在其他复发染色体区域的修饰剂映射到6Q（PHF10）和候选基因改造。这些变化在UM发展中的后果知之甚少。扩展和完善我们现在将进行DNA和RNA的长阅读单分子测序的拷贝数研究从具有不同的驱动突变和CNA的主要UMS分离。在目标1和2中，我们将进行长阅读单分子测序（SMRT）以开发基因组结构变化（SV）的详细图在这些肿瘤中，识别和表征关键断点，基因融合和肿瘤特异性同工型。除了获得对染色体改变产生的见解，我们还将验证和完善焦点删除并确定此类地区的关键同工型。细胞系中的功能分析将检查基因融合和同工型的后果与拷贝数变化高度相关。并行和瞄准 3我们将利用蝇遗传学的力量研究D的UM发生重大变化的结果。 Melanogaster。我们将首先生成和表征苍蝇，而BAP1直系同源物中功能突变的丧失 calypso，并产生GNAQ直系同源物GαQ的激活突变，以使丢失的综合作用可以研究CALYPSO和GαQ的激活。我们将专注于幼虫眼盘和机翼光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学这些突变引起的变化。飞行模型能够解决该功能相关性通过执行快速遗传相互作用，来自所有三个目标的基因组见解。将来，候选人UM司机从AIMS 1-2识别的将测试是否能够修改我们在两个热门单曲中看到的类似癌症的表型 GαQ和Calypso飞行模型。过度生长，组织转化和转移的遗传补充表型将代表令人兴奋的治疗候选者，以在以后的研究中追求。