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

开发针对小细胞肺癌的 ASCL1 和 NeuroD1 谱系癌基因靶向治疗

基本信息

批准号：
10240702
负责人：
JOHN D. MINNA
金额：
$ 60.51万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-25 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10240702
关键词：
3-Dimensional ASCL1 gene Ablation Advocate BETA2 protein Behavior Biological Assay Biological Markers CRISPR/Cas technology Cancer Model Cancer Patient Cancer cell line Cell Line Cells ChIP-seq Chromatin Clinical Collection Congresses Data Data Set Databases Dependence Drops Drug usage Enrollment Family member Frequencies Future Genes Genetic Genetically Engineered Mouse Growth Health Human In Vitro Individual Induction of Apoptosis Knock-out Knowledge Libraries Malignant - descriptor Malignant Neoplasms Methodology Methods Modeling Molecular Molecular and Cellular Biology Mus Mutate NFIB gene Neoplasm Circulating Cells Neurosecretory Systems Oncogenes Pathogenesis Patients Pattern Pharmaceutical Preparations Pharmacology Phenotype Pre-Clinical Model Prevention approach Prevention therapy Proteins Publishing Regulator Genes Research Personnel Resistance Role Specimen Technology Testing Therapeutic Therapeutic Effect Translations Work Xenograft Model Xenograft procedure base behavior in vitro cancer survival cancer therapy cell killing clinical translation clinically translatable druggable target in vitro Assay in vivo knock-down lung small cell carcinoma mRNA Expression molecular marker neoplastic cell new therapeutic target novel therapeutics patient derived xenograft model pre-clinical precision medicine predicting response predictive marker protein expression resistance frequency resistance mechanism response skills small hairpin RNA success targeted treatment therapeutic development therapy development transcription factor transcriptome sequencing tumor

项目摘要

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.

开发针对小细胞肺癌 (SCLC) 的 ASCL1 和 NEUROD1 谱系癌基因靶向治疗该申请重点开发针对 SCLC 的新靶向疗法，重点关注两个关键谱系癌基因 ASCL1 和 NEUROD1 参与 SCLC 发病机制和恶性行为，近 90% 的 SCLC 均参与其中。在临床前模型中表达 ASCL1、NEUROD1 或两者，包括人类 SCLC 系和异种移植物。 SCLC 基因工程小鼠模型 (GEMM)，SCLC 是表达 ASCL1 或 NEUROD1 的肿瘤似乎对其表达和功能“上瘾”，ASCL1 或 NEUROD1 的存在也与之相关。如果 ASCL1/NEUROD1 被去除，则具有重要的下游癌基因和调节基因的表达。（通过基因敲除）小细胞肺癌许多都使用最先进的技术进行肿瘤细胞杀伤记录。人类临床前模型，我们建议系统地研究大量 SCLC 的依赖性细胞系和异种移植物（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）通过基因敲低对ASCL1和NEUROD1进行阻断，并系统测试阻断能力这两个转录因子的遗传和药理学下游潜在“可成药”靶标我们有三个具体目标：目标 1. 确定 ASCL1 和 NEUROD1 表达模式以及临床前 SCLC 模型和肿瘤标本的临床和分子相关性目标 2. 确定 ASCL1 和 NEUROD1 遗传依赖性表型，预测反应的潜在分子生物标志物， SCLC 临床前模型中耐药的频率和机制；目标 3. 确定 ASCL1 的作用； NEUROD1 直接调节“下游”目标作为可用于治疗的漏洞使用体内异种移植 shRNA 迷你文库“退出”筛选和选定的抑制下游药物的效果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。使用 CRISPR-CAS9 技术开发 SCLC 疗法，包括其潜在机制耐药性，我们将探索 ASCL1 和 NEUROD1 表达作为 SCLC 入组的可能性为“精准医学”开发的生物标记物可预测此类靶向治疗在个体中的反应 SCLC 已开发出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人类 SCLC 细胞系集合，异种移植物，以及 SCLC 的重要 GEMM，2) 生成直接监管的完整列表通过 ChipSeq/RNASeq 和染色质景观研究了解 ASCL1 和 NEUROD1 的下游靶标， 3) 系统研究 SCLC 对 ASCL1 依赖性的实验开发方法我们组建了一支世界一流的研究团队，其中包括一名患者。倡导者，具有互补的技能，以确保该项目的成功完成。将作为 SCLC 新 ASCL1 和 NEUROD1 靶向治疗的基础。