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

描述（由申请人提供）：我们已经通过在冷泉港实验室（CSHL）开发的高分辨率基因组谱分析方法完成了对超过250个肺癌样本的分析。我们发现了12个主要的重复扩增子，其中大部分包含已知的驱动基因，如MYC或KRAS 2，还有三个不包含，包括位于14q13的第二个最常见的扩增子。我们的第一个具体目标是在14q13这个频繁扩增的区域内对所有三个候选驱动基因进行全面的功能评估。这种全面的功能评价将利用获得的功能转化测定和功能丧失测定，使用新的RNA干扰技术，在CSHL开发，在体外和体内沉默基因表达。我们有初步的数据表明，所有三个基因，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，我们还建立了合作关系，以解决14q13扩增的潜在诊断和预后意义。另外两个新的频繁扩增子包含几个候选基因。为了解决含有许多过表达基因的扩增子的功能分析中固有的困难，我们的第二个具体目标是开发新的条形码化RNA文库干扰技术，该技术能够并行地对许多基因进行功能分析，成为用于具有大量候选驱动基因的扩增子的功能分析的强大工具。我们的研究结果将对肺癌发生的分子基础产生新的见解，并应确定这种高致死性癌症的关键治疗靶点和诊断新策略。本研究的第一个目标是发现和功能表征肺癌最致命形式之一的新致病基因。第二个目标是创建一个基于基因功能的工具，使研究人员能够快速识别包含太多基因的复发性扩增基因组区域内的致病基因，以通过逐个基因的方法进行研究。该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.