Graduate Institute of Oncology, National Taiwan University College of Medicine, Taipei, 10051, Taiwan.
Graduate Institute of Toxicology, National Taiwan University College of Medicine, Taipei, 10051, Taiwan.
Theranostics. 2021 Sep 21;11(19):9667-9686. doi: 10.7150/thno.62676. eCollection 2021.
The tumorous niche may drive the plasticity of heterogeneity and cancer stemness, leading to drug resistance and metastasis, which is the main reason of treatment failure in most cancer patients. The aim of this study was to establish a tumor microenvironment (TME)-based screening to identify drugs that can specifically target cancer stem cells (CSCs) and cancer-associated fibroblasts (CAFs) in the TME. Lung cancer patient-derived cancer cell and CAFs were utilized to mimic the TME and reproduce the stemness properties of CSCs and develop a high-throughput drug screening platform with phenotypical parameters. Limiting dilution assay, sphere-forming and ALDH activity assay were utilized to measure the cancer stemness characteristics. patient-derived xenograft (PDX) models and single-cell RNA sequencing were used to evaluate the mechanisms of the compounds in CSCs and CAFs. The TME-based drug screening platform could comprehensively evaluate the response of cancer cells, CSCs and CAFs to different treatments. Among the 1,524 compounds tested, several drugs were identified to have anti-CAFs, anticancer and anti-CSCs activities. Aloe-emodin and digoxin both show anticancer and anti-CSCs activity and , which was further confirmed in the lung cancer PDX model. The combination of digoxin and chemotherapy improved therapeutic efficacy. The single-cell transcriptomics analysis revealed that digoxin could suppress the CSCs subpopulation in CAFs-cocultured cancer cells and cytokine production in CAFs. The TME-based drug screening platform provides a tool to identify and repurpose compounds targeting cancer cells, CSCs and CAFs, which may accelerate drug development and therapeutic application for lung cancer patients.
肿瘤微环境可能驱动异质性和癌症干性的可塑性，导致耐药性和转移，这是大多数癌症患者治疗失败的主要原因。本研究旨在建立一种基于肿瘤微环境（TME）的筛选方法，以鉴定能够特异性靶向 TME 中的癌症干细胞（CSC）和癌症相关成纤维细胞（CAF）的药物。利用肺癌患者来源的癌细胞和 CAF 模拟 TME，重现 CSC 的干性特征，并开发具有表型参数的高通量药物筛选平台。有限稀释分析、球体形成和 ALDH 活性分析用于测量癌症干性特征。患者来源的异种移植（PDX）模型和单细胞 RNA 测序用于评估化合物在 CSC 和 CAF 中的作用机制。基于 TME 的药物筛选平台可以全面评估癌症细胞、CSC 和 CAF 对不同治疗的反应。在测试的 1524 种化合物中，有几种药物被鉴定具有抗 CAF、抗癌和抗 CSC 的活性。大黄素和地高辛均显示出抗癌和抗 CSC 的活性，这在肺癌 PDX 模型中得到了进一步证实。地高辛与化疗联合应用可提高治疗效果。单细胞转录组学分析显示，地高辛可以抑制 CAF 共培养的癌细胞中的 CSC 亚群和 CAF 中的细胞因子产生。基于 TME 的药物筛选平台为鉴定和重新利用靶向癌细胞、CSC 和 CAF 的化合物提供了一种工具，可能加速肺癌患者的药物开发和治疗应用。
