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

 描述(申请人提供)：肺癌是美国和全世界与癌症相关的死亡的主要原因(Herbst等人)。2008年)。非小细胞肺癌(NSCLC)，约占所有肺癌病例的85%。20-30%的人类NSCLC肿瘤获得了抗氧化剂转录因子Nrf2(功能增益(GOF))或其负调控因子Keap1(LOF)的突变，这表明在肺癌发生过程中，氧化应激稳态在维持癌细胞存活方面发挥了重要作用。尽管在这一途径中观察到了高频率的突变，但人们对其在肺癌发生和发展中的作用知之甚少。为了研究非小细胞肺癌，我们的实验室利用了非小细胞肺癌的基因工程小鼠模型(GEMM)，该模型忠实地概括了人类非小细胞肺癌的组织学、分子和进展特征。肿瘤在原地(天然)组织环境中发展，在完整的肿瘤微环境中发展，在没有混淆的烟草诱变剂的情况下发展。我最近开发了一种快速而精确的体内方法，它省去了为设计感兴趣的新等位基因而对小鼠胚系进行耗时操作的需要。我将复杂的基于CRE/loxP的GEMM的能力与高精度的基因组编辑CRISPR/Cas9系统结合在一起。使用这一强大的新方法，我将使用NSCLC GEMM从功能上研究Nrf2和Keap1在肺腺癌发生和发展中的肿瘤细胞自主作用。这项申请K22 NIH职业过渡奖的研究重点是通过结合遗传和生化方法的严格体内实验方法来阐明Nrf2抗氧化途径在启动和进展中的作用。这些定义明确且可实现的实验的目标和时间表是：*剖析Nrf2在非小细胞肺癌肿瘤发生中的功能重要性*确定Nrf2在肺癌中介导的代谢变化*确定对调节其体内抗氧化功能至关重要的Nrf2靶点*为在NSCLC GEMM和人类细胞系中观察到的实验串扰提供平台，这对未来的研究将被证明是有价值的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以及更重要的是我在泰勒·贾克斯博士的实验室中正在进行的努力所塑造的，我正在努力揭示这一本地非小细胞肺癌小鼠模型的启动和进展所涉及的基本机制。我受益于麻省理工学院贾克斯实验室和周边地区的研究环境，那里提供了无与伦比的科学讨论、合作和培训机会。麻省理工学院、博德研究所和哈佛医学院的科学界提供了无数的研讨会和工作坊，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。这是一次令人难以置信的经历，它将赋予我在该奖项的独立阶段及以后管理实验室所需的许多技能。这项提议代表了我致力于通过联合合作者的分析和使用复杂的基因工程小鼠来识别非小细胞肺癌肿瘤发生的根本驱动因素的科学投资。我打算启动一个独立的研究项目，利用我从活体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提供坚实的基础。我期待着教育和招收和我一样热爱癌症研究的学生和博士后。

 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.