New Therapeutic Targets in Small Cell Lung Cancer that are Epistatic or Synthetic Lethal with pRB Loss

项目摘要/摘要 小细胞肺癌(SCLC)是一种高度恶性的神经内分泌癌，缺乏有效的靶向治疗。 识别小细胞肺癌的治疗靶点一直是具有挑战性的，部分原因是小细胞肺癌的驱动突变 主要是功能丧失，涉及肿瘤抑制基因RB1和TP53。而TP53基因高度突变 在许多成人实体肿瘤中，RB1(蛋白pRb)几乎普遍缺失是小细胞肺癌所特有的。我提议 通过研究与pRb的上位性和合成致死性相互作用来确定小细胞肺癌的新治疗靶点 损失，前者是作用于PRB下游的目标，后者代表创建的依赖关系 具体地说就是公关部门的损失。我使用了这两种策略来确定两个新的目标，KDM5A和 AURKB是小细胞肺癌增殖所必需的，其特定功能是调节神经内分泌 分化和染色体分离。在目标1中，我将使用生化和遗传 确定KDM5A调节ASCL1的具体机制的方法 小细胞肺癌肿瘤形成所需的神经内分泌谱系转录激活剂。在目标2中，基于 我从CRISPR筛查得到的初步数据，我假设染色体分离的其他调节者 是具有RB1缺失的合成致命性的。我将使用基于直接荧光的竞争分析来验证真的 人工合成的致命相互作用。然后我将使用时间推移显微镜来确定潜在的机制，通过 其中pRb缺失加剧了由其他染色体分离基因缺失引起的有丝分裂表型。在……里面 目的3，我将询问KDM5A是否是肿瘤启动和/或体内肿瘤维持所必需的 我开发的新的基于CRISPR的小鼠小细胞肺癌基因工程模型。 我是一名具有癌症生物学研究背景的内科肿瘤学家，正在申请K08奖项 长期目标是成为一名终身教职的独立实验室研究员。我设想开发一种 调查小细胞肺癌发病机制的独立研究计划 最终目标是为小细胞肺癌患者寻找新的治疗方法。在我提议的K08研究培训期间，我将 在达纳-法伯癌症研究所(DFCI)威廉·凯林博士的实验室进行指导研究。我 我计划把90%的时间花在研究上，10%的时间花在病人护理上，去看胸部肿瘤病人。我有过 组织了一个杰出的咨询委员会，帮助指导我与教职员工的研究和职业发展 DFCI、哈佛医学院和麻省理工学院的成员，他们是我建议的特定领域的专家 研究包括：斯图尔特·奥尔金博士、大卫·佩尔曼博士、泰勒·贾克斯博士和斯蒂芬·布莱克洛博士。布鲁斯博士 约翰逊是世界著名的肺癌临床试验专家，他将担任我的临床顾问。这位杰出的 DFCI的环境加上课程和会议将帮助我实现我的长期目标 职业抱负。

Project Summary/Abstract Small cell lung cancer (SCLC) is a high-grade neuroendocrine cancer with no effective targeted therapies. Identifying therapeutic targets in SCLC has been challenging, partly because driver mutations in SCLC are primarily loss of function, involving the tumor suppressor genes RB1 and TP53. While TP53 is highly mutated in many adult solid tumors, the near-universal loss of RB1 (protein pRB) is unique to SCLC. I propose to identify new therapeutic targets in SCLC by investigating epistatic and synthetic lethal interactions with pRB loss, with the former being targets acting downstream of pRB and the latter representing dependencies created specifically by pRB loss. I have employed both of these strategies to identify two novel targets, KDM5A and AURKB, that are required for SCLC proliferation and whose specific function is to regulate neuroendocrine differentiation and chromosomal segregation, respectively. In Aim 1, I will use biochemical and genetic approaches to determine the specific mechanism by which KDM5A regulates ASCL1, a pulmonary neuroendocrine lineage transcriptional activator that is required for SCLC tumor formation. In Aim 2, based on my preliminary data from a CRISPR screen, I hypothesize that other regulators of chromosomal segregation are synthetic lethal with RB1 loss. I will use direct fluorescence-based competition assays to validate true synthetic lethal interactions. I will then use time-lapse microscopy to determine the underlying mechanism by which pRB loss exacerbates the mitotic phenotype caused by loss of other chromosomal segregation genes. In Aim 3, I will ask whether KDM5A is necessary for tumor initiation and/or tumor maintenance in vivo using a novel CRISPR-based genetically-engineered mouse model of SCLC that I developed. I'm a medical oncologist with a research background in cancer biology applying for a K08 award with a long-term goal of becoming a tenure-track independent laboratory investigator. I envision developing an independent research program investigating the mechanisms responsible for SCLC pathogenesis with the ultimate goal of identifying new therapies for SCLC patients. During my proposed K08 research training, I will perform mentored research in the laboratory of Dr. William Kaelin at the Dana-Farber Cancer Institute (DFCI). I plan to spend 90% of my time on research and 10% on patient care seeing thoracic oncology patients. I have organized an outstanding advisory committee to help guide my research and career development with faculty members at DFCI, Harvard Medical School, and MIT that are experts in specific areas of my proposed research including: Dr. Stuart Orkin, Dr. David Pellman, Dr. Tyler Jacks, and Dr. Stephen Blacklow. Dr. Bruce Johnson, a world-renowned lung cancer clinical trialist, will serve as my clinical advisor. This outstanding environment at DFCI supplemented with coursework and conferences will help me achieve my long-term career aspirations.