Somatic Mutation Discovery in Lung Squamous Cell Carcinoma

肺鳞状细胞癌中体细胞突变的发现

• 批准号: 8018168
• 负责人: Alexis Hiram Ramos
• 金额: $ 2.9万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8018168
• 关键词: Accounting; Anchorage-Independent Growth; Biological Assay; Calculi; Cancer Etiology; Cancer Patient; Cell Line; Complementary DNA; DNA; DNA Resequencing; DNA Sequence Analysis; Data; Dideoxy Chain Termination DNA Sequencing; Epithelial; Frequencies; Future; Genes; Genome; Genomics; Goals; Imatinib; In Vitro; Lead; Lung; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; MicroRNAs; Modeling; Molecular Profiling; Mutate; Mutation; Mutation Spectra; NIH 3T3 Cells; Pathogenesis; Patients; Pharmacotherapy; Phosphotransferases; Play; Point Mutation; RNA Interference; Role; Sampling; Shotgun Sequencing; Somatic Mutation; Squamous Cell; Squamous Cell Lung Carcinoma; Squamous cell carcinoma; Techniques; Technology; Testing; bcr-abl Fusion Proteins; cancer therapy; fusion gene; interest; mutant; novel; overexpression; research study; small hairpin RNA; tumor; tumorigenesis; tumorigenic

DESCRIPTION (provided by applicant): The goal of this proposal is to discover novel cancer-causing somatic mutations in lung squamous cell carcinoma (SCC) with a combination of multiple sequencing technologies and functional experiments. Lung SCC accounts for 30% of all lung cancer cases. Unlike other types of lung cancer, there are no targeted therapies available for lung SCC patients. It is hoped that an accurate characterization of the spectrum of mutations in Lung SCC will lead to new avenues of rational drug therapy in lung SCC. Aim 1: Determine frequency of newly discovered somatic mutations in lung SCC. In preliminary experiments, I analyzed DNA sequence data for 201 genes and 496 microRNAs in 20 lung SCC samples. This study discovered 27 somatic point mutations across 17 different genes. Examination of the mutations revealed several potentially interesting findings. In order to determine which genes are recurrently mutated in lung squamous cancer and thus likely to be involved in tumorigenesis, Sanger sequencing of select mutated genes will be performed in an expanded set of 96 primary lung SCC samples. Aim 2: Utilize single-template Solexa sequencing for discovery of fusion genes. Fusion genes often drive malignancy in many hematological cancers. Importantly, fusion genes such as BCR-ABL can be effectively treated with imatinib. Recent findings in other cancers suggest that fusion genes could also play a significant role in epithelial cancers. TO discover fusion genes in lung SCC, I will perform Solexa shotgun sequencing of cDNA in lung SCC cell line models. Aim 3: Validate mutation and fusion gene candidates from Aims 1 and 2 with in vitro transformation assays. In order to determine which alterations from Aims 1 and 2 are truly involved in pathogenesis, I will systematically test discovered mutations and fusion genes with transformation assays and RNAi experiments in in vitro cell line models. This proposal will attempt to discover mutations in lung cancer patients by analyzing DNA from tumor samples. It is hoped that newly found mutations will provide the stepping stone for future lung cancer therapies.

描述（由申请人提供）：该提案的目的是在肺鳞状细胞癌（SCC）中发现新型引起癌症的体细胞突变，并结合了多个测序技术和功能实验。肺SCC占所有肺癌病例的30％。与其他类型的肺癌不同，没有针对肺SCC患者的靶向疗法。希望对肺SCC突变频谱的准确表征将导致肺SCC中有理学药物治疗的新途径。目标1：确定肺SCC中新发现的体细胞突变的频率。在初步实验中，我在20个肺SCC样品中分析了201个基因的DNA序列数据和496个microRNA。这项研究发现了17个不同基因的27个体细胞突变。对突变的检查发现了一些可能有趣的发现。为了确定哪些基因在肺鳞癌中反复突变，因此可能参与肿瘤发生，将在一组扩展的96个原发性肺SCC样品中进行选择的突变基因的Sanger测序。 AIM 2：利用单个板索莱克萨测序来发现融合基因。融合基因通常会在许多血液学癌症中引起恶性肿瘤。重要的是，可以用伊马替尼有效治疗诸如BCR-ABL之类的融合基因。其他癌症的最新发现表明，融合基因也可能在上皮癌中发挥重要作用。为了发现肺SCC中的融合基因，我将在肺SCC细胞系模型中对cDNA进行solexa shot弹枪测序。 AIM 3：从AIM 1和2中验证候选突变基因，并在体外转化测定中验证。为了确定AIM 1和2的哪些改变真正参与发病机理，我将系统地测试在体外细胞系模型中使用转化测定和RNAi实验的突变和融合基因。该建议将尝试通过分析肿瘤样品中的DNA来发现肺癌患者的突变。希望新发现的突变将为未来的肺癌疗法提供垫脚石。