Elucidating the role of the Nrf2 anti-oxidant pathway in lung adenocarcinoma

 描述（由申请人提供）：肺癌是美国和全球癌症相关死亡的主要原因（Herbst et al. 2008）。非小细胞肺癌（NSCLC）占所有肺癌病例的约85%。20-30%的人NSCLC肿瘤获得抗氧化转录因子Nrf 2（功能获得性（GOF））或其负调节因子Keap 1（LOF）的突变，表明氧化应激稳态在肺癌发生过程中维持癌细胞存活的重要作用。尽管在该途径中观察到的突变频率很高，但对其在肺肿瘤发生和进展中的作用知之甚少。为了研究NSCLC，我们的实验室利用NSCLC的基因工程小鼠模型（GEMM），该模型忠实地再现了人类NSCLC的组织学、分子学和进展特征。肿瘤在原位（天然）组织背景下，在存在完整肿瘤微环境且不存在混杂烟草诱变剂的情况下发生。我最近开发了一种快速而精确的体内方法，该方法绕过了为了工程化感兴趣的新等位基因而对鼠生殖系进行耗时操作的需要。我将复杂的基于Cre/loxP的GEMM与高度精确的基因组编辑CRISPR/Cas9系统相结合。使用这种强大的新方法，我将使用NSCLC GEMM功能研究Nrf 2和Keap 1在肺腺癌发生和进展中的肿瘤细胞自主作用。在K22 NIH职业过渡奖的申请中提出的研究重点是通过结合遗传和生物化学方法的严格体内实验方法阐明Nrf 2抗氧化途径在启动和进展中的作用。其中概述的这些明确定义且可实现的实验的目标和时间轴是：* 剖析Nrf 2在NSCLC肿瘤发生中的功能重要性 * 鉴定Nrf 2在肺癌中介导的代谢变化 * 鉴定对于介导其体内抗氧化功能重要的Nrf 2靶点 * 为NSCLC GEMM和人类细胞系中的观察结果之间的实验串扰提供平台，这将被证明对未来的研究有价值。本申请中描述的项目是由我对癌症生物学领域的长期奉献形成的，更重要的是，我在泰勒杰克斯博士的实验室中不断努力，以揭示在NSCLC的这种本地小鼠模型中涉及起始和进展的基本机制。我从麻省理工学院杰克实验室的研究环境中受益匪浅，周围地区为科学讨论，合作和培训提供了无与伦比的机会。麻省理工学院、布罗德研究所和哈佛医学院的科学界提供了无数的研讨会和讲习班，继续促进我的科学发展。此外，我还指导了三名麻省理工学院的本科生，三名暑期学生和两名技术助理，他们直接与我一起进行与拟议研究项目有关的实验。这是一个令人难以置信的经验，将赋予我许多必要的技能，在这个奖项的独立阶段和超越管理实验室。该提案代表了我对确定NSCLC肿瘤发生的基本驱动因素的专门科学投资，通过联合合作者的分析和使用复杂的基因工程小鼠。我打算启动一个独立的研究项目，利用我从体内动物模型中获得的洞察力，目标是将我的发现转化为人类系统。从长远来看，我相信这些实验将为我的研究计划奠定坚实的基础。我期待着教育和招募学生和博士后分享我对癌症研究的热情。

 DESCRIPTION (provided by applicant): Lung cancer is the leading cause of cancer-related deaths in the United States and worldwide (Herbst et al. 2008). Non-small-cell lung cancer (NSCLC), accounts for ~85% of all lung cancer cases. 20-30% of human NSCLC tumors acquire mutations in anti-oxidant transcription factor Nrf2 (gain-of-function (GOF)) or its negativ regulator Keap1 (LOF) suggesting an important role for oxidative stress homeostasis to maintain cancer cell survival during lung carcinogenesis. Despite the high frequency of mutations observed in this pathway, little is know about its role in lung tumor initiation and progression. To study NSCLC our laboratory utilizes a genetically engineered mouse model (GEMM) of NSCLC that faithfully recapitulates the histologic, molecular, progression features of human NSCLC. Tumors develop in the autochthonous (native) tissue context, in the presence of an intact tumor microenvironment and in the absence of confounding tobacco mutagens. I have recently developed a rapid and precise in vivo method that bypasses the need for time- consuming manipulation of the murine germline in order to engineer novel alleles of interest. I have combined the power of sophisticated Cre/loxP-based GEMMs with the highly precise genome editing CRISPR/Cas9 system. Using this powerful new approach I will functionally investigate the tumor cell- autonomous role of Nrf2 and Keap1 in the lung adenocarcinoma initiation and progression using the NSCLC GEMM. The research proposed within this application for a K22 NIH career transition award focuses on elucidating the role of the Nrf2 anti-oxidant pathway in initiation and progression by rigorous in vivo experimental approaches combining both genetic and biochemical approaches. The goals and timeline of these well-defined and achievable experiments outlined within are to: * Dissect the functional importance of Nrf2 in NSCLC tumorigenesis * Identify the metabolic changes mediated by Nrf2 in lung cancer * Identify the Nrf2 targets that are important for mediating its anti-oxidant functions in vivo * Provide a platform for experimental cross talk between observations in NSCLC GEMMs and human cell lines, which will prove valuable for future research The project described within this application has been shaped by my long-standing dedication to the field of cancer biology and more importantly my ongoing efforts in Dr. Tyler Jacks' laboratory to uncover the fundamental mechanisms involved in initiation and progression in this autochthonous mouse model of NSCLC. I have benefited from the research environment in the Jacks Laboratory, MIT, and the surrounding area that offers unmatched opportunities for scientific discussion, collaboration, and training. The scientific community at MIT, the Broad Institute, and Harvard Medical School offers countless seminars and workshops that continue to foster my scientific development. Additionally, I supervise have supervised three undergraduate MIT student, three summer students and two technical assistant that work directly with me on experiments pertaining to the proposed research project. This is an incredible experience that will endow me with many of the necessary skills to manage a laboratory during the independent phase of this award and beyond. This proposal represents my dedicated scientific investment towards identifying the fundamental drivers of NSCLC tumorigenesis, by uniting collaborators' analyses and the use of a sophisticated genetically engineered mouse. It is my intention to start an independent research program that will capitalize on the insight I obtain from the in vivo animal model with the goal to translate my findings in human systems. For the long-term, I am confident that these experiments will provide a solid foundation on which my research program can be built upon. I look forward to educating and recruiting students and postdocs that share my passion for cancer research.