Identification and characterization of driver gene(s) in recurrent lung cancer am

描述(申请人提供)：我们已经用冷泉港实验室(CSHL)开发的高分辨率基因组图谱方法完成了对250多个肺癌样本的分析。我们发现了12个主要的重复扩增，其中大多数包含已知的驱动基因，如MYC或KRAS2，另外3个不包含，包括位于14q13的第二频繁扩增。我们的第一个特定目标是在14q13对这个频繁扩增区域内的所有三个候选驱动基因进行全面的功能评估。这项全面的功能评估将利用CSHL在这里开发的新的RNA干扰技术，利用功能获得转化分析和功能丧失分析，在体外和体内沉默基因表达。我们有初步数据表明，这三个基因，每个编码不同的转录因子，协同促进肺上皮细胞的增殖。除了功能分析，我们还合作解决了14q13扩增的潜在诊断和预后意义。另外两个新的频繁扩增片段包含几个候选基因。为了解决包含许多过表达基因的扩增片段的功能分析固有的困难，我们的第二个具体目标是开发新的条形码RNA文库干扰技术，使多个基因的功能分析成为一种强大的工具，用于包含大量候选驱动基因的扩增片段的功能分析。我们的结果将对肺癌发生的分子基础产生新的见解，并将确定这种高度致命的癌症的关键治疗靶点和新的诊断策略。这项研究的第一个目标是发现最致命的肺癌之一的新的致病基因及其功能特征。第二个目标是创建一种基于基因功能的工具，使研究人员能够在重复扩增的基因组区域中快速识别原因基因，该基因组区域包含太多基因，无法通过逐个基因的方法进行研究。本研究方案的前瞻性研究进展有望对未来肺癌的诊断和治疗产生积极影响。

DESCRIPTION (provided by applicant): We have completed analysis of over 250 lung cancer samples by a high- resolution genome profiling method developed here at Cold Spring Harbor Laboratory (CSHL). We found twelve major recurrent amplicons, most of which contain known driver genes such as MYC or KRAS2, and three others that do not, including the second most frequent amplicon which is located at 14q13. Our first specific aim is to perform comprehensive functional evaluation of all three candidate driver genes within this frequently amplified region at 14q13. This comprehensive functional evaluation will utilize both gain-of-function transformation assays and loss-of-function assays using new RNA interference technology, developed here at CSHL, that silences gene expression in vitro and in vivo. We have preliminary data that indicates that all three genes, each encoding a different transcription factors, synergistically promote the proliferation of lung epithelial cells. In addition to functional analysis, we have formed collaborations to address the potential diagnostic and prognostic significance of 14q13 amplification. The other two novel frequent amplicons contain several candidate genes. To address the difficulty inherent in functional analysis of amplicons containing many overexpressed genes, our second specific aim is to develop new barcoded RNA library interference technology, which enables functional analysis of many genes in parallel, into a robust tool for functional analysis of amplicons with large numbers of candidate driver genes. Our results will produce new insights into the molecular basis of lung carcinogenesis and should identify key therapeutic targets and new strategies for diagnostics for this highly lethal cancer. The first goal of this study focuses on the discovery and functional characterization of novel causal genes of one of the most deadly forms of lung cancer. The second goal is to create a gene function-based tool to enable researchers to rapidly identify the causal gene within a recurrent amplified genomic region that contains too many genes to be studied by a gene-by-gene approach. Forward research progress of this research proposal is expected to positively impact the diagnosis and treatment of lung cancer in the future.