Functional analysis of genetic alterations occuring in uveal melanoma

葡萄膜黑色素瘤中发生的遗传改变的功能分析

• 批准号: 227076835
• 负责人: Privatdozent Dr. Klaus Georg Griewank
• 金额: --
• 依托单位: Klinik für Dermatologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/227076835?language=en
• 关键词: Functional; analysis; genetic; alterations; occuring

The experiments described in this proposal aim to analyze the genetic alterations found in uveal melanoma. The goal is to examine the function of the recently discovered gene mutations in GNAQ and GNA11 both in vitro and in vivo as well as establish corresponding mouse models.1) Characterization of GNAQ and GNA11 Q209L oncogenes in melanocytesThe function of the oncogenes GNAQ and GNA11 in the development of uveal melanomas is currently poorly understood. To achieve this, the biological function of this gene is to be studied in the following fashion: 1. Determine the protein domains and signal effectors that are essential for tumor formation, 2. A detailed analysis of GNAQ/11 oncogene induce protein-phosphorylations and RNA expression, 3. Analysis of GNAQ/11 induced senescence or apoptosis, 4. Studying the metastasis profile of GNAQ/11 oncogene transformed cells, 5. Participation of other genetic factors i.e. BAP1, MYC, PTEN and p53 in GNAQ/11 oncogene induced tumors and metastasis. The goal of this first project aim is to identify functional mechanisms that could be of potential use for developing effective therapeutic strategies.2) Establishing GNAQ/11 oncogene mouse modelsIn contrast to an extensive list of existing mouse models with various oncogenes (BRAF, HRAS, KIT) for cutaneous melanomas, there are currently no existing mouse models of GNAQ/11 oncogenes. Establishing a model of uveal melanoma using these physiological oncogenes would be valuable both from a basic science as well as a translational oncology perspective. With the design and cloning of various transgenic and knock-in constructs during his postdoc in the US, the applicant has laid the groundwork for this project. These mice will allow the expression of GNAQ/11 oncogenes at various timepoints in different tissues. It is planned to: 1. Test the effect of this expression, both on tumor genesis as well as embryonic development in both melanocytic and other tissue. 2. Through different crossings, analyze in vivo the role of additional gene alterations affecting i.e. BAP1, CDKN2A, p53, MYC and PTEN in combination with GNAQ/11 oncogene expression. 3. Apply the mouse model to test a variety of different therapeutic approaches for effectively inhibiting tumor growth and metastasis.

该提案中描述的实验旨在分析葡萄膜黑色素瘤中发现的遗传改变。目的是在体外和体内检查最近发现的GNAQ和GNA11基因突变的功能，并建立相应的小鼠模型。1）黑色素细胞中GNAQ和GNA11 Q209L癌基因的表征目前对癌基因GNAQ和GNA11在葡萄膜黑色素瘤发展中的功能知之甚少。为了实现这一目标，应通过以下方式研究该基因的生物学功能： 1. 确定肿瘤形成所必需的蛋白质结构域和信号效应子， 2. 详细分析 GNAQ/11 癌基因诱导的蛋白质磷酸化和 RNA 表达， 3. 分析 GNAQ/11 诱导的衰老或凋亡， 4. 研究 GNAQ/11 癌基因转化细胞的转移谱， 5.其他遗传因子BAP1、MYC、PTEN和p53参与GNAQ/11癌基因诱导的肿瘤和转移。第一个项目的目标是确定可能用于开发有效治疗策略的功能机制。2) 建立 GNAQ/11 癌基因小鼠模型与大量现有的具有各种皮肤黑色素瘤癌基因（BRAF、HRAS、KIT）的小鼠模型相比，目前尚无 GNAQ/11 癌基因的小鼠模型。从基础科学和转化肿瘤学的角度来看，使用这些生理癌基因建立葡萄膜黑色素瘤模型都是有价值的。申请人在美国博士后期间设计并克隆了各种转基因和敲入构建体，为该项目奠定了基础。这些小鼠将允许 GNAQ/11 癌基因在不同时间点在不同组织中表达。计划： 1. 测试该表达对肿瘤发生以及黑素细胞和其他组织中胚胎发育的影响。 2. 通过不同的杂交，结合 GNAQ/11 癌基因表达，体内分析影响 BAP1、CDKN2A、p53、MYC 和 PTEN 的其他基因改变的作用。 3.应用小鼠模型测试多种不同的治疗方法，以有效抑制肿瘤的生长和转移。