Pellinen Teijo, Paavolainen Lassi, Martín-Bernabé Alfonso, Papatella Araujo Renata, Strell Carina, Mezheyeuski Artur, Backman Max, La Fleur Linnea, Brück Oscar, Sjölund Jonas, Holmberg Erik, Välimäki Katja, Brunnström Hans, Botling Johan, Moreno-Ruiz Pablo, Kallioniemi Olli, Micke Patrick, Östman Arne
Institute for Molecular Medicine Finland, Helsinki Institute of Life Science, University of Helsinki, Helsinki, Finland.
Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden.
J Natl Cancer Inst. 2023 Jan 10;115(1):71-82. doi: 10.1093/jnci/djac178.
Cancer-associated fibroblasts (CAFs) are molecularly heterogeneous mesenchymal cells that interact with malignant cells and immune cells and confer anti- and protumorigenic functions. Prior in situ profiling studies of human CAFs have largely relied on scoring single markers, thus presenting a limited view of their molecular complexity. Our objective was to study the complex spatial tumor microenvironment of non-small cell lung cancer (NSCLC) with multiple CAF biomarkers, identify novel CAF subsets, and explore their associations with patient outcome.
Multiplex fluorescence immunohistochemistry was employed to spatially profile the CAF landscape in 2 population-based NSCLC cohorts (n = 636) using antibodies against 4 fibroblast markers: platelet-derived growth factor receptor-alpha (PDGFRA) and -beta (PDGFRB), fibroblast activation protein (FAP), and alpha-smooth muscle actin (αSMA). The CAF subsets were analyzed for their correlations with mutations, immune characteristics, and clinical variables as well as overall survival.
Two CAF subsets, CAF7 (PDGFRA-/PDGFRB+/FAP+/αSMA+) and CAF13 (PDGFRA+/PDGFRB+/FAP-/αSMA+), showed statistically significant but opposite associations with tumor histology, driver mutations (tumor protein p53 [TP53] and epidermal growth factor receptor [EGFR]), immune features (programmed death-ligand 1 and CD163), and prognosis. In patients with early stage tumors (pathological tumor-node-metastasis IA-IB), CAF7 and CAF13 acted as independent prognostic factors.
Multimarker-defined CAF subsets were identified through high-content spatial profiling. The robust associations of CAFs with driver mutations, immune features, and outcome suggest CAFs as essential factors in NSCLC progression and warrant further studies to explore their potential as biomarkers or therapeutic targets. This study also highlights multiplex fluorescence immunohistochemistry-based CAF profiling as a powerful tool for the discovery of clinically relevant CAF subsets.
癌症相关成纤维细胞（CAFs）是分子异质性的间充质细胞，与恶性细胞和免疫细胞相互作用，并具有抗肿瘤和促肿瘤功能。先前对人类CAFs的原位分析研究很大程度上依赖于对单一标志物进行评分，因此对其分子复杂性的认识有限。我们的目的是利用多种CAF生物标志物研究非小细胞肺癌（NSCLC）复杂的空间肿瘤微环境，识别新的CAF亚群，并探讨它们与患者预后的关联。
采用多重荧光免疫组织化学技术，使用针对4种成纤维细胞标志物的抗体，对2个基于人群的NSCLC队列（n = 636）中的CAF格局进行空间分析，这4种标志物分别为血小板衍生生长因子受体α（PDGFRA）和β（PDGFRB）、成纤维细胞活化蛋白（FAP）和α平滑肌肌动蛋白（αSMA）。分析CAF亚群与突变、免疫特征、临床变量以及总生存期的相关性。
两个CAF亚群，即CAF7（PDGFRA-/PDGFRB+/FAP+/αSMA+）和CAF13（PDGFRA+/PDGFRB+/FAP-/αSMA+），在肿瘤组织学、驱动基因突变（肿瘤蛋白p53 [TP53]和表皮生长因子受体[EGFR]）、免疫特征（程序性死亡配体1和CD163）以及预后方面显示出具有统计学意义的相反关联。在早期肿瘤（病理肿瘤-淋巴结-转移IA-IB期）患者中，CAF7和CAF13是独立的预后因素。
通过高内涵空间分析确定了多标志物定义的CAF亚群。CAFs与驱动基因突变、免疫特征和预后之间的密切关联表明，CAFs是NSCLC进展中的重要因素，值得进一步研究以探索它们作为生物标志物或治疗靶点的潜力。本研究还强调基于多重荧光免疫组织化学的CAF分析是发现临床相关CAF亚群的有力工具。
