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

项目1--项目摘要 尽管我们对癌症发病机制的理解取得了巨大的进步，但个体的治疗 接受常规化疗或靶向药物治疗的患者仍然具有高度的经验性。到目前为止，精度 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，并不是所有基于标记物的预测都能保证患者的反应，因为许多都是 从基于人口的研究中得出的相关性。利用个别患者肿瘤标本进行EX的方法 体内药敏试验同样受到产生细胞系的过程和随后的 对药物敏感性的影响。现有的检测癌细胞生长的方法，如基于三磷酸腺苷的生长分析 (CellTiter-Glo)，需要延长培养时间和大量肿瘤细胞。这就排除了它们用于疾病的可能性。 对大多数癌症患者进行监测。此外，这些成批的方法不适合描述 亚群内的治疗易感性。因此，迫切需要快速和便捷的方法。 为了表征单个肿瘤标本的治疗敏感性，这些标本捕获了异质性，并可以 适用于非常小的标本，包括最小的残留疾病。项目1利用了一套独特的工具 概述影响单个癌细胞对治疗干预反应的内在因素。 我们将询问配对的表型和转录测量在多大程度上可以识别出 调节细胞自主抵抗，并强调克服这种抵抗的治疗方法。癌 细胞将从两种白血病的原发肿瘤或患者来源的细胞系/异种移植中分离出来(作为 液体肿瘤模型)和结肠癌/胰腺癌(实体肿瘤模型)。与项目2不同，单元格将 与世隔绝地测量，不模仿微环境的各个方面。在几个小时的时间里，我们将 检查细胞的不同表型属性(质量和质量累积率)，并将这些属性联系起来 在单细胞水平上的转录组。然后，我们将通过以下方式确定细胞内在的抗性机制 分析有反应和无反应的肿瘤细胞的转录特征。

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.