Center for Cancer Research and Comprehensive Cancer Center, Medical University of Vienna, Borschkegasse 8a, 1090, Vienna, Austria.
Department of Analytical Chemistry, University of Vienna, Waehringer Straße 38, 1090, Vienna, Austria.
J Exp Clin Cancer Res. 2023 Jan 23;42(1):27. doi: 10.1186/s13046-022-02582-0.
Pleural mesothelioma (PM) is an aggressive malignancy with poor prognosis. Unlike many other cancers, PM is mostly characterized by inactivation of tumor suppressor genes. Its highly malignant nature in absence of tumor driving oncogene mutations indicates an extrinsic supply of stimulating signals by cells of the tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs) are an abundant cell type of the TME and have been shown to drive the progression of several malignancies. The aim of the current study was to isolate and characterize patient-derived mesothelioma-associated fibroblasts (Meso-CAFs), and evaluate their impact on PM cells.
Meso-CAFs were isolated from surgical specimens of PM patients and analyzed by array comparative genomic hybridization, next generation sequencing, transcriptomics and proteomics. Human PM cell lines were retrovirally transduced with GFP. The impact of Meso-CAFs on tumor cell growth, migration, as well as the response to small molecule inhibitors, cisplatin and pemetrexed treatment was investigated in 2D and 3D co-culture models by videomicroscopy and automated image analysis.
Meso-CAFs show a normal diploid genotype without gene copy number aberrations typical for PM cells. They express CAF markers and lack PM marker expression. Their proteome and secretome profiles clearly differ from normal lung fibroblasts with particularly strong differences in actively secreted proteins. The presence of Meso-CAFs in co-culture resulted in significantly increased proliferation and migration of PM cells. A similar effect on PM cell growth and migration was induced by Meso-CAF-conditioned medium. Inhibition of c-Met with crizotinib, PI3K with LY-2940002 or WNT signaling with WNT-C59 significantly impaired the Meso-CAF-mediated growth stimulation of PM cells in co-culture at concentrations not affecting the PM cells alone. Meso-CAFs did not provide protection of PM cells against cisplatin but showed significant protection against the EGFR inhibitor erlotinib.
Our study provides the first characterization of human patient-derived Meso-CAFs and demonstrates a strong impact of Meso-CAFs on PM cell growth and migration, two key characteristics of PM aggressiveness, indicating a major role of Meso-CAFs in driving PM progression. Moreover, we identify signaling pathways required for Meso-CAF-mediated growth stimulation. These data could be relevant for novel therapeutic strategies against PM.
胸膜间皮瘤（PM）是一种预后不良的侵袭性恶性肿瘤。与许多其他癌症不同，PM 主要表现为肿瘤抑制基因失活。在没有肿瘤驱动致癌基因突变的情况下，其高度恶性表明肿瘤微环境（TME）中的细胞提供了刺激信号。癌症相关成纤维细胞（CAFs）是 TME 中丰富的细胞类型，并已被证明可促进多种恶性肿瘤的进展。本研究的目的是分离和鉴定患者来源的间皮瘤相关成纤维细胞（Meso-CAFs），并评估其对 PM 细胞的影响。
从 PM 患者的手术标本中分离出 Meso-CAFs，并通过 array 比较基因组杂交、下一代测序、转录组学和蛋白质组学进行分析。将 GFP 逆转录病毒转导到人的 PM 细胞系中。通过视频显微镜和自动图像分析，在 2D 和 3D 共培养模型中研究 Meso-CAF 对肿瘤细胞生长、迁移以及对小分子抑制剂顺铂和培美曲塞治疗的反应的影响。
Meso-CAFs 显示正常的二倍体基因型，没有 PM 细胞中典型的基因拷贝数异常。它们表达 CAF 标志物，缺乏 PM 标志物表达。它们的蛋白质组和分泌组谱与正常肺成纤维细胞明显不同，特别是在主动分泌蛋白方面有很大差异。在共培养中存在 Meso-CAF 会导致 PM 细胞的增殖和迁移显著增加。Meso-CAF 条件培养基也会对 PM 细胞的生长和迁移产生类似的影响。在不影响 PM 细胞的浓度下，用克唑替尼抑制 c-Met、LY-2940002 抑制 PI3K 或 WNT-C59 抑制 WNT 信号传导，可显著抑制共培养中 Meso-CAF 介导的 PM 细胞生长刺激。Meso-CAF 不能保护 PM 细胞免受顺铂的影响，但对 EGFR 抑制剂厄洛替尼有明显的保护作用。
本研究首次对人源患者来源的 Meso-CAFs 进行了特征描述，并证明了 Meso-CAFs 对 PM 细胞生长和迁移的强烈影响，这是 PM 侵袭性的两个关键特征，表明 Meso-CAFs 在推动 PM 进展中起着重要作用。此外，我们确定了 Meso-CAF 介导的生长刺激所需的信号通路。这些数据可能对针对 PM 的新型治疗策略具有重要意义。
