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 – 项目概要 尽管我们对癌症发病机制的理解取得了巨大进步，但个体治疗 接受传统化疗或靶向药物的患者仍具有高度经验性。迄今为止，精度 肿瘤学方面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 瘤。然而，并非所有基于标记的预测都能保证患者的反应，因为许多预测都是基于标记的预测的结果 基于人群的研究的相关性。利用个体患者肿瘤标本进行治疗的方法 体内药物敏感性测试同样受到细胞系生成过程和后续过程的限制。 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)，需要扩展培养和大量肿瘤细胞。这排除了它们用于治疗疾病的可能性 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 亚人群内的治疗敏感性。因此，迫切需要快速且简便的方法 表征个体肿瘤样本中的治疗敏感性，捕获异质性并且可以 适用于非常小的标本，包括微小残留病。项目 1 利用一套独特的工具 分析影响个体癌细胞对治疗干预反应的内在因素。 我们将询问配对表型和转录组测量在多大程度上可以识别以下途径： 介导细胞自主抵抗并强调克服该抵抗的治疗方法。癌症 细胞将从原发性肿瘤或两种白血病的患者来源的细胞系/异种移植物中分离出来（作为 液体肿瘤模型）和结肠癌/胰腺癌（作为实体瘤模型）。与项目 2 相比，单元格将 单独测量而不模仿微环境的各个方面。在几个小时的时间内，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 单细胞水平的转录组。然后我们将通过以下方式确定细胞内在的耐药机制： 分析有反应和无反应肿瘤细胞的转录组特征。