Tumor-barcoding coupled with high-throughput sequencing of a novel chemoradiation resistant SCLC mouse model

肿瘤条形码与新型放化疗耐药 SCLC 小鼠模型的高通量测序相结合

• 批准号: 10201805
• 负责人: CHRISTINE L. HANN
• 金额: $ 15.35万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-03 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201805
• 关键词: ASCL1 gene; Animal Model; Animals; Bar Codes; Biological Models; Cancer Model; Cancer Therapy Evaluation Program; Candidate Disease Gene; Cell Line; Cells; Chemoresistance; Chest; Cisplatin; Collaborations; Communities; Coupled; Data; Development; Epithelial; Etiology; Etoposide; Experimental Models; Funding; Gene Expression; Genetic; Genetically Engineered Mouse; Genotype; Goals; Harmine; High-Throughput Nucleotide Sequencing; Human; Imaging Device; In Vitro; Letters; Limited Stage; Link; Logistics; Lung Neoplasms; Malignant Neoplasms; Maps; Measures; Medical; Medical Oncologist; Mesenchymal; Modeling; Molecular; Mutate; Natural History; Non-Small-Cell Lung Carcinoma; Oncology; Organoids; Outcome; Parents; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacogenetics; Pharmacogenomics; Pharmacology; Radiation; Radiation Oncologist; Radiation therapy; Recurrence; Recurrent disease; Regulator Genes; Research Design; Resistance; Role; Sampling; Subgroup; Target Populations; Techniques; Testing; Time; Tissues; Transitional Epithelium; Tumor Suppressor Proteins; Veterinarians; Work; base; candidate validation; chemoradiation; chemotherapy; deep sequencing; genetic approach; image guided; improved; in vivo; innovation; insight; lung small cell carcinoma; molecular subtypes; mouse model; novel; patient derived xenograft model; patient population; patient stratification; personalized medicine; pre-clinical; programs; research and development; resistance mechanism; response; therapy resistant; tool; transcription factor; treatment effect; treatment response; tumor; tumor barcoding and sequencing; tumor growth; tumorigenesis; vector

PROJECT ABSTRACT Limited stage small cell lung cancer (LS SCLC), which is the only curable form of SCLC, is remarkably sensitive to etoposide plus cisplatin combined with radiation (XRT) with response rates of 90%, but recurrence is nearly universal. LS SCLC has only a 20-25% 5-year overall survival and ultimately a strategy for improving long-term outcome in SCLC needs to successfully target the population of cells within tumors destined to survive through standard therapy to inevitably give rise to recurrent disease. However, there is a considerable gap in understanding the specific mechanisms responsible for chemoradiation (chemoXRT) resistance in SCLC. Our project is unique among the current portfolio of SCLC funded programs in that we have focused on chemoXRT resistance to increase cure rates in LS SCLC. Recently, our work has suggested using patient- derived xenograft (PDX) models of SCLC may be an important tool to elucidate these mechanisms of chemoresistance. This approach identified a strongly synergistic anti-SCLC interaction that led to an approved CTEP trial based on our preclinical data - (NCI #10070: PI Hann). Our parent U01 had three Specific Aims: (1) Characterize natural history of response of experimental models of SCLC to chemoXRT in vivo. – where we proposed to determine response rates and recurrence natural histories from a panel of PDXs and available SCLC genetically engineered mouse models (GEMMs); (2) Characterization of molecular underpinnings of SCLC chemoXRT resistance – where we proposed constructing gene regulatory networks and gene expression profiles associated with chemoXRT resistance; and , (3) Pharmacologic and genetic validation of candidate genes for SCLC chemoXRT resistance in vitro and in vivo – where we proposed to validate novel gene candidates for conferring chemoXRT resistance using pharmacologic and genetic approach with SCLC PDX-derived organoids and available SCLC GEMMs. In this revision proposal, we propose additional Specific Aims that are highly complementary to the parent U01 objectives but are completely novel. SCLC has recently been stratified into four molecular subgroups. Currently, there are no GEMMs that reliably represent the SCLC-Y (or -YAP1) subtype and using our novel epithelial-mesenchymal transition (EMT) GEMMs we propose to create a novel SCLC GEMM for the community. Using novel inducible Twist1 and SNAI1 mouse models we will examine whether Twist1 (or SNAI1) can accelerate autochthonous SCLC tumor development and are casual for chemoXRT resistance. Lastly, our revision proposal leverages a new technique, tumor barcoding with barcode deep-sequencing (Tuba-seq), to overcome all of the barriers that limited the feasibility of using SCLC GEMMs for quantitative pharmacogenomic analysis. We will use Tuba-seq, an innovative technique enabling lineage tracing and direct quantification of chemoXRT treatment effects on different genetic backgrounds in vivo to profile the pharmcogenetic landscape of SCLC.

项目摘要 局限期小细胞肺癌(LS SCLC)是目前唯一可治愈的小细胞肺癌。 对依托泊苷和顺铂联合放射治疗(XRT)敏感，有效率为90%，但复发 几乎是普遍存在的。LS SCLC的5年总存活率只有20%-25%，并最终制定了改善战略 小细胞肺癌的长期结果需要成功地靶向肿瘤内的细胞群体 通过标准治疗存活下来，不可避免地会引发复发的疾病。然而，有相当大的 在对化疗耐药的具体机制的理解上存在差距 SCLC。我们的项目在目前SCLC资助的项目组合中是独一无二的，因为我们专注于 化疗XRT耐药可提高LS SCLC的治愈率。最近，我们的工作建议使用患者- 小细胞肺癌衍生异种移植(PDX)模型可能是阐明这些机制的重要工具。 化疗耐药。这种方法发现了一种强烈的协同抗小细胞肺癌相互作用，导致了批准的 基于我们临床前数据的CTEP试验-(NCI#10070：Pi Hann)。我们的母公司U01有三个具体目标：(1) 描述实验性小细胞肺癌模型对体内化疗XRT反应的自然历史。-我们在哪里 建议从一组PDX中确定应答率和复发自然病史，并提供 小细胞肺癌基因工程小鼠模型(GEMM)；(2)小细胞肺癌的分子基础 小细胞肺癌化学XRT耐药--我们建议构建基因调控网络和基因 与化疗XRT耐药性相关的表达谱；以及，(3)药理学和遗传学验证 小细胞肺癌化疗XRT耐药候选基因的体内外研究 用药理学和遗传学方法研究小细胞肺癌化疗XRT耐药候选基因 PDX衍生的有机化合物和可用的SCLC GEMM。在这项修订建议中，我们提出了额外的具体建议 目标与母公司的U01目标高度互补，但完全是新奇的。SCLC最近 被分成四个分子亚群。目前，还没有可靠地代表 SCLC-Y(或-YAP1)亚型和使用我们的新型上皮-间充质转化(EMT)GEMM 为社区打造一个新颖的SCLC GEMM。利用新型诱导型Twist1和SNAI1小鼠模型 将研究Twist1(或SNAI1)是否可以加速固有的小细胞肺癌肿瘤的发展 对化疗XRT耐药的随意。最后，我们的修订方案利用了一种新技术--肿瘤条形码 使用条形码深度测序(Tuba-Seq)，以克服限制使用的可行性的所有障碍 用于定量药物基因组分析的SCLC GEMMS。我们将使用Tuba-seq，一种创新的技术 实现谱系追踪和直接量化化疗XRT对不同基因的影响 活体研究背景描述小细胞肺癌的药物遗传学图景。