Investigating the role of KLF5 genomic alterations in tumorigenesis

项目摘要 博士Xiaoyang Zhang是Dana的Matthew Meyerson博士实验室的博士后研究员， 法伯癌症研究所和布罗德研究所。他的长期职业目标，符合使命的 NCI通过确定癌症发展的机制来降低癌症相关的死亡率和痛苦 并确定有吸引力的治疗靶点。为了实现这一目标，张博士独特地利用了 基因组学和细胞和分子生物学方法来回答与癌症有关的基本问题 生物学 人类癌症的大规模基因组调查最近发现了两种类型的基因组改变， KLF 5基因。张博士发现，KLF 5基因附近的一个非编码区充满了超级- 增强子，在几种癌症类型中局部扩增，并且扩增与KLF 5过度相关。 表情此外，Meyerson博士的实验室发现了DNA结合中的重要热点突变， 非小细胞肺癌中KLF 5基因的结构域。所有这些都有力地表明，KLF 5是一个新颖的 癌基因该建议旨在描述这些基因组改变在调节KLF 5活性中的作用 和促进肿瘤发生。目的1研究KLF 5基因的超级增强子扩增与KLF 5基因的关系 过度表达，以及KLF 5过度表达在不同癌症类型中的致癌后果。然后， 目的2：利用KLF 5野生型同基因突变体，研究KLF 5热点突变的致癌功能， 通过基因组编辑工具产生的突变细胞系。最后，在目标3中，CRISPR介导的启动子/增强子 将进行筛选以鉴定KLF 5突变体的功能结合位点及其相关靶点 基因和途径。这项拟议中的研究将提高我们对致癌转录如何 这些因素促进癌症发展，并可能产生新的治疗靶点。 博士张将在生物统计学，计算生物学和基因组编辑方面获得更多的研究培训， 同时通过赠款撰写和领导能力方面的培训加强其职业发展。的 优秀的研究环境和设施提供给张博士，包括实验室空间和充分的 丹娜-法伯癌症研究所和布罗德研究所的机构准入。在K99阶段，博士。 张的研究和培训将在Matthew Meyerson博士的主要指导下进行， 癌症基因组学的领导者，并将通过与计算专家的合作进一步加强 生物学和高通量遗传筛选，以及由咨询委员会指导， 迈尔斯·布朗、布拉德利、伯恩斯坦、张峰和李维·加罗威博士。

PROJECT SUMMARY Dr. Xiaoyang Zhang is a postdoctoral research fellow in the laboratory of Dr. Matthew Meyerson at Dana- Farber Cancer Institute and the Broad Institute. His long-term career goal, in alignment with the mission of the NCI, is to reduce cancer-associated mortality and suffering by determining mechanisms of cancer development and identifying attractive therapeutic targets. To accomplish this goal, Dr. Zhang uniquely leverages both genomics and cellular and molecular biology methods to answer fundamental questions relating to cancer biology. Large-scale genomic surveys of human cancer have recently found two types of genomic alterations of the KLF5 gene. Dr. Zhang identified that a non-coding region, nearby the KLF5 gene and filled with super- enhancers, is focally amplified in several cancer types and the amplification correlates with KLF5 over- expression. In addition, Dr. Meyerson's laboratory identified significant hot-spot mutations in the DNA binding domain of the KLF5 gene in non-small cell lung cancers. All these strongly suggest that KLF5 is a novel oncogene. This proposal aims to characterize the role of these genomic alterations in regulating KLF5 activity and promoting tumorigenesis. Aim 1 will study the link between the super-enhancer amplification and KLF5 over-expression, and the oncogenic consequences of KLF5 over-expression in diverse cancer types. Then, Aim 2 will study the oncogenic function of KLF5 hot-spot mutations by using isogenic KLF5 wild-type and mutant cell lines generated by genome-editing tools. Finally, in Aim 3, a CRISPR-mediated promoter/enhancer screening will be developed to identify the functional binding sites of KLF5 mutants and their associated target genes and pathways. The proposed research will improve our understanding of how oncogenic transcription factors promote cancer development and may yield novel therapeutic targets. Dr. Zhang will gain more research training in biostatistics, computational biology and genome-editing, and simultaneously enhance his career development through training in grant-writing and leadership. The outstanding research environment and facilities available to Dr. Zhang include laboratory space and full institutional access at both Dana-Farber Cancer Institute and the Broad Institute. During the K99 phase, Dr. Zhang's research and training will be carried out under the primary mentorship of Dr. Matthew Meyerson, a leader in cancer genomics, and will be additionally enhanced by collaborations with experts in computational biology and high-throughput genetic screening, as well as by mentoring from an advisory committee consisting of Drs. Myles Brown, Bradley Bernstein, Feng Zhang, and Levi Garraway.