Functional landscape of Eph receptor mutations in melanoma

黑色素瘤中 Eph 受体突变的功能景观

基本信息

批准号：
8638686
负责人：
ELENA B PASQUALE
金额：
$ 25.45万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-01 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8638686
关键词：
Affect Agreement Amino Acid Sequence BRAF gene Binding Biological Biological Markers Biological Process CDKN2A gene Cancer cell line Cataloging Catalogs Cell Culture Techniques Cell Line Cell Proliferation Cell surface Cells Collection Computer Simulation Data Development Eph Family Receptors EphA Receptors EphA3 Receptor EphB4 Receptor Ephrin B Receptor Ephrins Exposure to Family Frequencies Future Gene Family Gene Mutation Genes Genomics Growth Hot Spot Human Human Genome Ligand Binding Ligands Link MEKs Malignant - descriptor Malignant Neoplasms Measures Melanoma Cell Metastatic Melanoma Missense Mutation Modeling Mutate Mutation Nonsense Mutation Oncogenic PTB Domain PTEN gene Pathogenesis Pathologic Processes Pathway interactions Patients Phosphorylation Phosphotransferases Play Positioning Attribute Property Protein Tyrosine Kinase Proteins Proto-Oncogene Proteins c-akt Receptor Protein-Tyrosine Kinases Receptor Signaling Recurrence Research Role Sampling Signal Transduction Site Skin Somatic Mutation Specimen Splice-Site Mutation Structure Subfamily lentivirinae Surface Surveys TP53 gene Tumor Cell Line Tumor Suppressor Proteins Tyrosine Phosphorylation Ultraviolet Rays cancer cell cancer type design exome genome sequencing in vivo melanocyte melanoma member migration mutant novel outcome forecast public health relevance receptor receptor binding receptor function tool trafficking tumor

项目摘要

DESCRIPTION (provided by applicant): Among all cancers, metastatic melanoma is one of the most deadly. In most cases melanoma develops due to multiple gene mutations induced in skin melanocytes by exposure to UV light, but few of these mutations have been studied in detail. Most melanomas harbor mutations that constitutively activate the RAS-RAF-MEK-ERK pathway, such as the frequent NRAS and BRAF mutations, which are well known to play a critical role in melanoma malignancy. However, mutations in other pathways such as the PTEN, TP53 or CDKN2A tumor suppressors are also needed for melanoma development. Furthermore, recent studies have uncovered new mutations in other gene families that contribute to melanoma malignancy. The objective of the proposed research is to explore the novel hypothesis that Eph receptor mutations play a functional role in melanoma development and/or progression. Recent sequencing studies of ~300 melanoma specimens and cell lines have revealed that approximately half of the tumors carry one or more nonsynonymous mutations in genes of the Eph receptor tyrosine kinase family. The Eph receptors regulate many fundamental biological and pathological processes, such as the growth and invasiveness of cancer cells, and several of them have been implicated in melanoma pathogenesis. Various in silico analyses predict that many of the Eph receptor mutations contribute to malignancy, consistent with our preliminary studies showing that several EphA2 and EphA3 melanoma mutations analyzed so far drastically affect receptor functional properties. We therefore propose two aims to explore the functional significance of Eph mutations in melanoma. In Aim 1, we will characterize a subset of the mutations identified to determine if they affect the abilit of Eph receptors to perform their normal functions, including trafficking to the cell surface, binding ephrin ligands, assembling into receptor signaling clusters, and fulfilling their role as tyrosine kinases. We will especially focus on Eph receptors that are either known or very likely to play a role in melanoma pathogenesis and on the mutations most likely to be "driver" mutations. In Aim 2, we will investigate how representative Eph mutations affect melanoma cell development and/or malignancy by introducing mutant and wild-type receptors in melanocytes and melanoma cells, and measuring cell proliferation/survival and migration/invasiveness. Surveying the functional effects of Eph receptor mutations occurring in melanoma will provide valuable information on how melanoma cells acquire their malignant properties and on the involvement of the Eph receptor family in cancer development and progression, which is currently incompletely understood. This information, and the collection of research tools to be generated, will also enable future more extensive studies to characterize the importance of normal as well as mutated Eph receptors in additional cell culture and in vivo melanoma models. Cataloguing the effects of Eph gene mutations may also prove useful for identifying novel biomarkers as well as genomic alterations to help prognosis and the design of individualized therapies for melanoma patients.

描述（由申请人提供）：在所有癌症中，转移性黑色素瘤是最致命的一种。在大多数情况下，黑色素瘤由于暴露于紫外线而诱导的多种基因突变而出现，但详细研究了这些突变中的其中很少。大多数黑色素瘤均可激活Ras-Raf-Mek-ERK途径，例如频繁的NRA和BRAF突变，这些突变在众所周知，它们在黑色素瘤恶性肿瘤中起着至关重要的作用。但是，在黑色素瘤发育中，还需要其他途径中的突变，例如PTEN，TP53或CDKN2A肿瘤抑制子。此外，最近的研究发现了其他基因家族中有助于黑色素瘤恶性肿瘤的新突变。拟议的研究的目的是探索新的假设，即EPH受体突变在黑色素瘤发育和/或进展中起功能作用。最近对〜300个黑色素瘤标本和细胞系的测序研究表明，大约一半的肿瘤在EPH受体酪氨酸激酶家族的基因中携带一个或多个非同义突变。 EPH受体调节许多基本的生物学和病理过程，例如癌细胞的生长和侵入性，其中一些与黑色素瘤的发病机理有关。各种硅分析中的各种表明，许多EPH受体突变导致恶性肿瘤，这与我们的初步研究一致，表明到目前为止分析了几种EPHA2和EPHA3黑色素瘤突变，迄今为止对受体功能的影响很大。因此，我们提出了两个旨在探讨黑色素瘤中EPH突变的功能意义的目标。在AIM 1中，我们将表征一部分突变的子集，以确定它们是否影响EPH受体的影响，以执行其正常功能，包括运输到细胞表面，结合Ephrin配体，组装成受体信号簇，并履行其作为酪氨酸激酶的作用。我们将特别专注于已知或很可能在黑色素瘤发病机理和最有可能是“驱动器”突变的突变中发挥作用的EPH受体。在AIM 2中，我们将通过在黑色素细胞和黑色素瘤细胞中引入突变体和野生型受体以及测量细胞增殖/存活以及迁移/侵入性来研究代表性EPH突变如何影响黑色素瘤细胞发育和/或恶性肿瘤。调查黑色素瘤中发生的EPH受体突变的功能作用将提供有关黑色素瘤细胞如何获得其恶性特性以及EPH受体家族参与癌症发展和进展的有价值的信息，目前尚不完全了解这一点。这些信息以及要生成的研究工具的收集也将使未来的更广泛的研究能够表征正常和突变的EPH受体在其他细胞培养和体内黑色素瘤模型中的重要性。分类EPH基因突变的作用也可能被证明可用于鉴定新型生物标志物以及基因组改变，以帮助预后和针对黑色素瘤患者的个性化疗法设计。