Glioblastoma Genome Project to Locate Molecular Targets

描述（由申请人提供）：这项工作的主要目标是定位和评估脑癌的分子靶点，从胶质母细胞瘤开始。这个项目是更大的癌症重测序项目的试点。我们与文特尔研究所及其联合技术中心合作，利用他们的SNP检测管道进行最先进的低成本测序。对于我们的第一个特定目标，我们将在30个胶质母细胞瘤的癌症基因组中重新排序所有已知的激酶结构域。一旦检测到激酶结构域突变，我们将分析扩展到另外50个胶质母细胞瘤和其他脑癌的整个基因，包括儿童胶质母细胞瘤和髓母细胞瘤。在我们对前40个激酶基因的初步分析中，在FGFR1和PDGFRA中发现了新的酪氨酸受体激酶突变。我们还在成人和儿童胶质母细胞瘤中发现了PIK3CA的其他突变。在具体目标2中，我们建议使用数字核型技术创建30个胶质母细胞瘤的高分辨率拷贝数图，这样我们就可以在相同的样本中评估点突变、扩增和缺失。我们已经完成了8个胶质母细胞瘤的数字核型分析，并且先前发表了使用这种强大的技术在髓母细胞瘤中发现发育基因基因组扩增的文章。在特定目标3中，我们将对频率大于10%的突变进行功能评估，从我们已经在胶质母细胞瘤中发现的两个激酶突变开始。我们将在细胞系中创建突变的模型系统，并确定磷酸化改变的目标，以及细胞周期、细胞凋亡和/或入侵是否被改变。为了确保快速报告并将我们的工作整合到更大的努力中，我们建议为我们的第四个也是最后一个具体目标建立一个在线数据库和报告。胶质母细胞瘤的存活率很低，需要新的治疗方法。我们的长期目标是从系统分析中选择最佳的分子靶点，并确定抑制这些新突变是否将是一种成功的治疗策略。

DESCRIPTION (provided by applicant): The main goal of this work is to locate and evaluate molecular targets for brain cancers, starting with glioblastoma. This project serves as a pilot for larger cancer re-sequencing projects. We have teamed with the Venter Institute and its associated Joint Technology Center to perform state-of-the-art and low cost sequencing using their SNP detection pipeline. For our first specific aim, we will re-sequence all known kinase domains in the cancer genomes of 30 glioblastomas. Once a kinase domain mutation is detected we expand our analysis to the entire gene in 50 additional glioblastomas and a panel of other brain cancers, including pediatric glioblastomas and medulloblastomas. In our preliminary analysis of the first 40 kinase genes, novel tyrosine receptor kinase mutations were found in FGFR1 and PDGFRA. We have also found additional mutations in PIK3CA in adult and pediatric glioblastoma. In specific aim 2, we propose to create a high-resolution copy number map of our 30 glioblastomas using Digital Karyotyping, so we can evaluate in the same samples point mutations, amplifications and deletions. We have completed Digital Karyotyping in 8 glioblastomas, and previously published on using this powerful technique to find a developmental gene genomic amplification in medulloblastoma. In specific aim 3 we will functionally evaluate the mutations we find with frequency greater than 10%, starting with two kinase mutations we have already found in glioblastoma. We will create a model system of the mutation in cell lines and determine the target of altered phosphorylation and if the cell-cycle, apoptosis and/or invasion are altered. To ensure rapid reporting and integration of our work into larger efforts we propose an online database and reporting for our fourth and final specific aim. Glioblastomas have poor survival and new treatments are needed. Our long-term goal is to choose the best molecular targets from this systematic analysis and determine if inhibition of these new mutations will be a successful therapeutic strategy.