Developing ASCL1 and NeuroD1 lineage oncogene targeted therapy for small cell lung cancer

ASCL1和NeUROD1系癌基因靶向治疗小细胞肺癌的研究 该应用专注于开发新的针对小细胞肺癌的靶向治疗，重点关注两个关键的家系癌基因 ASCL1和NEUROD1参与了小细胞肺癌的发病机制和恶性行为。近90%的SCLC 快递ASCL1和/或NEUROD1。在临床前模型中，包括人小细胞肺癌细胞系和异种移植以及 小细胞肺癌的基因工程小鼠模型(GEM)，表达ASCL1或NEUROD1的肿瘤 表现出对它们的表达和功能“上瘾”。ASCL1或NEUROD1的存在也与 重要的下游癌基因和调控基因的表达。如果删除ASCL1/NEUROD1 (通过基因敲除)小细胞肺癌经历了许多肿瘤细胞杀伤的日志。使用最先进的技术在 人类临床前模型，我们建议系统地研究大量小细胞肺癌的依赖性 系和异种移植物(包括患者来源的异种移植物、PDX和循环肿瘤细胞来源的异种移植物， CDXs)对ASCL1和NeUROD1进行基因敲除，并系统检测其阻断能力 这两个转录因子的基因和药理下游潜在的“可用药”靶标 杀死小细胞肺癌。我们有三个具体的目标：目标1.确定ASCL1和NEUROD1的表达模式 临床前小细胞肺癌模型和肿瘤标本中的临床和分子相关性；目的2.确定 ASCL1和NeUROD1遗传依赖表型，预测反应的潜在分子生物标志物， 目的3.确定ASCL1在小细胞肺癌临床前模型中的作用 而NEUROD1直接调控的“下游”靶标是可用于治疗的漏洞 使用体内异种shRNA微库“退出”筛选和选定抑制下游药物的效果 “可下药”的目标。作为这些目标的一部分，我们还将确定是否针对ASCL1或NEUROD1的耐药性 利用CRISPR-Cas9技术开发小细胞肺癌的治疗方法，包括潜在的机制 耐药性，我们将探索ASCL1和NEUROD1表达作为SCLC登记的可能性 发展“精准医学”的生物标记物，以预测这种靶向治疗的个体疗效 SCLC。我们已经开发了本应用程序所基于的大量初步数据，包括1) 汇集了世界上最大的临床和分子注释的人类小细胞肺癌株系和 异种移植物以及小细胞肺癌的重要GEMM，2)生成直接受调控的 通过ChipSeq/RNAseq和染色质景观研究ASCL1和NeUROD1的下游靶标， 3)发展实验方法，系统研究小细胞肺癌对ASCL1和ASCL1的依赖性 NEUORD1下游目标。我们已经组建了一支世界级的调查团队，其中包括一名患者 倡导，具有互补的技能，以确保这一项目的成功完成。最终交付成果 将作为小细胞肺癌新的ASCL1和NEUROD1靶向治疗的基础。

Developing ASCL1 and NEUROD1 lineage oncogene targeted therapy for small cell lung cancer (SCLC) This application focuses on developing new targeted therapy for SCLC focusing on two key lineage oncogenes involved in SCLC pathogenesis and malignant behavior, ASCL1 and NEUROD1. Nearly 90% of SCLCs express ASCL1, NEUROD1 or both. In the preclinical models, including human SCLC lines and xenografts and genetically engineered mouse models (GEMMs) of SCLC, tumors that express either ASCL1 or NEUROD1 appear “addicted” to their expression and function. The presence of ASCL1 or NEUROD1 also are associated with expression of important downstream oncogenes and regulatory genes. If ASCL1/NEUROD1 are removed (through genetic knockdown) SCLCs undergo many logs of tumor cell kill. Using state of the art technology in human preclinical models, we propose to systematically study the dependency of a large number of SCLC lines and xenografts (including patient derived xenografts, PDXs, and circulating tumor cell derived xenografts, CDXs) on ASCL1 and NEUROD1 through genetic knockdown, and systematically test the ability of blocking genetically and pharmacologically downstream potentially “druggable” targets of these two transcription factors to kill SCLCs. We have three specific aims: Aim 1. Determine ASCL1 and NEUROD1 expression patterns and clinical and molecular correlates in preclinical SCLC models and tumor specimens; Aim 2. Determine ASCL1 and NEUROD1 genetic dependency phenotypes, potential molecular biomarkers predicting response, and frequency and mechanisms of resistance in SCLC preclinical models; Aim 3. Determine the role of ASCL1 and NEUROD1 directly regulated “downstream” targets as vulnerabilities that can be exploited for therapeutic effect using in vivo xenograft shRNA mini-library “drop out” screens and selected drugs that inhibit downstream “druggable” targets. As part of these aims we will also determine if resistance to ASCL1 or NEUROD1 targeted therapy in SCLCs develops using CRISPR-CAS9 technology including potential mechanisms of this resistance, and we will explore the possible use of ASCL1 and NEUROD1 expression as SCLC enrollment biomarkers for developing “precision medicine” to predict the response of such targeted therapy in individual SCLCs. We have developed a large amount of preliminary data on which this application is based including 1) assembling the world’s largest collection of clinically and molecularly annotated human SCLC lines and xenografts, as well as important GEMMs of SCLC, 2) generating a comprehensive list of directly regulated downstream targets of ASCL1 and NEUROD1 through ChipSeq/RNASeq and chromatin landscape studies, and 3) developing experimental approaches to systematically study the dependency of SCLCs on ASCL1 and NEUORD1 downstream targets. We have assembled a world class team of investigators, including a patient advocate, with complementary skills to assure the successful completion of this project. The final deliverables will serve as the basis for new ASCL1 and NEUROD1 targeted therapeutics for SCLC.