Project 1: Systematic discovery of cell-intrinsic mechanisms of cancer drug resistance

项目1：系统发现癌症耐药的细胞内在机制

• 批准号: 10162308
• 负责人: SCOTT R MANALIS
• 金额: $ 65.24万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-07 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10162308
• 关键词: Acute Myelocytic Leukemia; Biological Assay; Biopsy; Bone Marrow; Cancer Cell Growth; Cell Line; Cell division; Cells; Colon; Colon Carcinoma; Colonic Neoplasms; Disease; Drug Exposure; Exposure to; Expression Profiling; Genes; Genetic Markers; Growth; Heterogeneity; Hour; Human; In Situ; Individual; Intrinsic factor; Link; Liquid substance; Malignant Neoplasms; Malignant neoplasm of pancreas; Measurement; Measures; Mediating; Medical; Modeling; Molecular; Monitor; Oncology; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Population Study; Predisposition; Primary Neoplasm; Process; Proteins; RNA; Residual Neoplasm; Resistance; Solid Neoplasm; Specimen; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Tissue Sample; Tumor Volume; Validation; Xenograft procedure; base; cancer cell; cancer drug resistance; cancer type; cell growth; cell type; chemotherapy; drug sensitivity; effective therapy; epigenetic marker; genome-wide; genome-wide analysis; in vivo; individual patient; individual response; leukemia; mRNA Expression; neoplastic cell; new therapeutic target; novel; pancreatic neoplasm; patient response; physical property; precision medicine; resistance mechanism; response; single-cell RNA sequencing; targeted agent; therapy resistant; tool; transcriptome; transcriptomics; treatment response; tumor; tumor microenvironment

Project 1 – Project Summary Despite tremendous advances in our understanding of cancer pathogenesis, the treatment of individual patients with either conventional chemotherapy or targeted agents remains highly empiric. To date, precision medicine efforts in oncology have focused primarily on genetic or epigenetic biomarkers within an individual tumor. However, not all marker-based predictions guarantee patient response, as many are the result of correlations from population-based studies. Approaches that utilize individual patient tumors specimens for ex vivo drug susceptibility testing are similarly limited by the process of generating a cell line and subsequent effects on drug sensitivity. Existing assays that measure cancer cell growth, such as ATP-based growth assays (CellTiter-Glo), require extended culture and a large volume of tumor cells. This precludes their use for disease monitoring in most patients with cancer. Furthermore, these bulk approaches are ill-suited for characterizing therapeutic susceptibility within subpopulations. Thus, there is a pressing need for rapid and facile approaches to characterize therapeutic sensitivity within individual tumor specimens that capture heterogeneity and can be applied to very small specimens, including minimal residual disease. Project 1 leverages a unique suite of tools to profile the intrinsic factors that inform the responses of individual cancer cells to therapeutic interventions. We will ask to what extent paired phenotypic and transcriptomic measurements can identify pathways that mediate cell autonomous resistance and highlight therapeutic approaches to overcome that resistance. Cancer cells will be isolated from primary tumors or from patient-derived cell lines/xenografts of both leukemias (as a liquid tumor model) and colon/pancreatic cancers (as a solid tumor model). In contrast to Project 2, cells will be measured in isolation without mimicking aspects of the microenvironment. Over a period of many hours, we will examine distinct phenotypic attributes of the cells (mass and mass accumulation rate) and link these attributes to the transcriptome at the single-cell level. We will then determine cell intrinsic mechanisms for resistance by analyzing transcriptomic features of responding and non-responding tumor cells.

项目1 -项目摘要 尽管我们对癌症发病机制的理解有了巨大的进步，但个体化治疗仍然是一个难题。 接受常规化疗或靶向药物治疗的患者仍然高度依赖经验。迄今为止， 肿瘤学中的医学努力主要集中在个体内的遗传或表观遗传生物标志物上 肿瘤然而，并非所有基于标记物的预测都能保证患者的反应，因为许多预测都是基于 基于人口的研究的相关性。利用个体患者肿瘤标本进行 体内药物敏感性试验同样受到产生细胞系和随后的 对药物敏感性的影响。测量癌细胞生长的现有测定，例如基于ATP的生长测定 （CellTiter-Glo）需要延长的培养和大量的肿瘤细胞。这就排除了它们用于疾病 对大多数癌症患者进行监测。此外，这些批量方法不适合表征 亚群内的治疗敏感性。因此，迫切需要快速和简便的方法， 表征单个肿瘤标本内的治疗敏感性，这些肿瘤标本捕获异质性， 适用于非常小的标本，包括微小残留病变。项目1利用了一套独特的工具 分析告知个体癌细胞对治疗干预的反应的内在因素。 我们将询问配对的表型和转录组学测量在多大程度上可以识别 介导细胞自主抗性，并强调克服该抗性的治疗方法。癌 细胞将从两种白血病的原发性肿瘤或患者来源的细胞系/异种移植物（作为 液体肿瘤模型）和结肠/胰腺癌（作为实体肿瘤模型）。与项目2相比， 在不模仿微环境的情况下单独测量。在几个小时内，我们将 检查细胞的不同表型属性（质量和质量积累率），并将这些属性联系起来 在单细胞水平上与转录组相关联。然后，我们将通过以下方式确定细胞内在的耐药性机制： 分析应答和非应答肿瘤细胞的转录组学特征。