Single cell functional dissection of tumor microenviornment-driven drug resistance

项目摘要 实体瘤不仅含有癌细胞，还含有非恶性细胞，如成纤维细胞、内皮细胞、成纤维细胞和成纤维细胞。 细胞和巨噬细胞。近年来，多方面的证据表明，这些非 恶性细胞共同构成了“肿瘤微环境”，在疾病中起着至关重要的作用 进展和治疗反应。具体来说，非恶性肿瘤中关键信号通路的激活 微环境的细胞可以使附近的肿瘤细胞对抗癌药物产生抗性。但我们的 对控制微环境驱动的抗性的途径的了解非常有限 事实上，不存在方法来进行大规模的药理学或遗传学筛选， 微环境为了应对这一挑战，我们最近开发了一种基于微流体的平台， 使非恶性和恶性细胞能够在数千个微型芯片上并行共培养， 公寓在这个提议中，我们描述了一种系统地激活关键信号通路的策略， 非恶性细胞使用混合的慢病毒cDNA文库，然后评估这些扰动的影响 对药物敏感性的影响。具体来说，我们将使用此策略来映射 微环境信号通路，控制乳腺癌细胞对批准和 新兴的靶向治疗，然后使用集成的 实验和计算流水线。因此，该项目将产生第一个大规模的筛选平台， 细胞-细胞相互作用的功能解剖，并定义关键的，临床相关的微环境途径 形成抗癌药物反应的机制。

Project Summary Solid tumors contain not only cancer cells, but also non-malignant cells such as fibroblasts, endothelial cells, and macrophages. In recently years, multiple lines of evidence have made it clear that these non- malignant cells, which together comprise the “tumor microenvironment”, play crucial roles in disease progression and therapeutic response. Specifically, the activation of key signaling pathways in non-malignant cells of the microenvironment can render nearby tumor cells resistant to anticancer drugs. However, our understanding of the landscape of pathways that govern microenvironment-driven resistance is highly limited by the fact that no methods exist to perform large-scale pharmacological or genetic screening of the microenvironment. To address this challenge, we recently developed a microfluidics-based platform that enables the parallel co-culture of non-malignant and malignant cells in thousands of miniature, on-chip “apartments”. In this proposal, we describe a strategy of systematically activating key signaling pathways in non-malignant cells using pooled lentiviral cDNA libraries, then assessing the impacts of these perturbations on drug sensitivity in nearby breast cancer cells. Specifically, we will use this strategy to map the microenvironmental signaling pathways that control the sensitivity of breast cancer cells to approved and emerging targeted therapies, then credential and validate hits from these assays using an integrated experimental and computational pipeline. Thus, this project will yield the first screening platform for large-scale functional dissection of cell-cell interactions and define key, clinically relevant microenvironmental pathways that shape anticancer drug responses.