Functional landscape of Eph receptor mutations in melanoma

黑色素瘤中 Eph 受体突变的功能景观

• 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
• 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

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(申请人提供)：在所有癌症中，转移性黑色素瘤是最致命的癌症之一。在大多数情况下，黑色素瘤的发生是由于紫外线照射导致皮肤黑素细胞发生多种基因突变所致，但很少有人对这些突变进行详细研究。大多数黑色素瘤都含有结构性地激活RAS-RAF-MEK-ERK通路的突变，例如频繁的NRAS和BRAF突变，这是众所周知的在黑色素瘤恶性中起关键作用的突变。然而，其他途径的突变，如PTEN、TP53或CDKN2A肿瘤抑制基因也是黑色素瘤发生所必需的。此外，最近的研究发现了导致黑色素瘤恶性的其他基因家族的新突变。这项研究的目的是探索一种新的假设，即Eph受体突变在黑色素瘤的发生和/或进展中发挥功能作用。最近对大约300个黑色素瘤标本和细胞系的测序研究表明，大约一半的肿瘤携带Eph受体酪氨酸激酶家族基因的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，如癌细胞的生长和侵袭，其中一些与黑色素瘤的发病机制有关。各种电子分析预测，许多Eph受体突变会导致恶性肿瘤，这与我们的初步研究一致，该研究表明，到目前为止分析的几个EphA2和EphA3黑色素瘤突变会严重影响受体的功能特性。因此，我们提出了两个目的来探索Eph突变在黑色素瘤中的功能意义。在目标1中，我们将描述已确定的突变的一个子集，以确定它们是否影响Eph受体执行其正常功能的能力，包括运输到细胞表面，结合ePhin配体，组装成受体信号簇，以及发挥其作为酪氨酸激酶的作用。我们将特别关注已知的或很可能在黑色素瘤发病机制中发挥作用的Eph受体，以及最有可能成为“驱动”突变的突变。在目标2中，我们将通过在黑素细胞和黑色素瘤细胞中引入突变型和野生型受体，并测量细胞的增殖/存活和迁移/侵袭性，来研究具有代表性的Eph突变如何影响黑色素瘤细胞的发育和/或恶性程度。研究发生在黑色素瘤中的Eph受体突变的功能效应将为了解黑色素瘤细胞如何获得其恶性特性以及Eph受体家族在癌症发生发展中的作用提供有价值的信息，目前对此尚不完全清楚。这些信息，以及即将产生的研究工具的收集，也将使未来更广泛的研究能够表征正常和突变的Eph受体在额外的细胞培养和体内黑色素瘤模型中的重要性。编目Eph基因突变的影响也可能被证明有助于识别新的生物标记物以及基因组变化，以帮助黑色素瘤患者的预后和个性化治疗的设计。