Akimova Tatiana, Zhang Tianyi, Negorev Dmitri, Singhal Sunil, Stadanlick Jason, Rao Abhishek, Annunziata Michael, Levine Matthew H, Beier Ulf H, Diamond Joshua M, Christie Jason D, Albelda Steven M, Eruslanov Evgeniy B, Hancock Wayne W
Division of Transplant Immunology, Department of Pathology and Laboratory Medicine, and Biesecker Center for Pediatric Liver Diseases, Children's Hospital of Philadelphia and Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Division of Thoracic Surgery, Department of Surgery, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
JCI Insight. 2017 Aug 17;2(16). doi: 10.1172/jci.insight.94075.
Experimental data indicate that FOXP3+ Tregs can markedly curtail host antitumor immune responses, but the properties of human intratumoral Tregs are still largely unknown, in part due to significant methodologic problems. We studied the phenotypic, functional, epigenetic, and transcriptional features of Tregs in 92 patients with non-small-cell lung cancer, comparing the features of Tregs within tumors versus corresponding blood, lung, and lymph node samples. Intratumoral Treg numbers and suppressive function were significantly increased compared with all other sites but did not display a distinctive phenotype by flow cytometry. However, by undertaking simultaneous evaluation of mRNA and protein expression at the single-cell level, we demonstrated that tumor Tregs have a phenotype characterized by upregulated expression of FOXP3 mRNA and protein as well as significantly increased expression of EOS, IRF4, SATB1, and GATA1 transcription factor mRNAs. Expression of these "Treg-locking" transcription factors was positively correlated with levels of FOXP3 mRNA, with highest correlations for EOS and SATB1. EOS had an additional, FOXP3 mRNA-independent, positive correlation with FOXP3 protein in tumor Tregs. Our study identifies distinctive features of intratumoral Tregs and suggests that targeting Treg-locking transcription factors, especially EOS, may be of clinical importance for antitumor Treg-based therapy.
实验数据表明，FOXP3+调节性T细胞（Tregs）可显著抑制宿主抗肿瘤免疫反应，但人类肿瘤内Tregs的特性仍很大程度上未知，部分原因是存在重大的方法学问题。我们研究了92例非小细胞肺癌患者中Tregs的表型、功能、表观遗传和转录特征，比较了肿瘤内Tregs与相应血液、肺和淋巴结样本中Tregs的特征。与所有其他部位相比，肿瘤内Treg数量和抑制功能显著增加，但通过流式细胞术未显示出独特的表型。然而，通过在单细胞水平同时评估mRNA和蛋白质表达，我们证明肿瘤Tregs具有以FOXP3 mRNA和蛋白质表达上调以及EOS、IRF4、SATB1和GATA1转录因子mRNA表达显著增加为特征的表型。这些“Treg锁定”转录因子的表达与FOXP3 mRNA水平呈正相关，其中EOS和SATB1的相关性最高。在肿瘤Tregs中，EOS与FOXP3蛋白存在另外一种不依赖于FOXP3 mRNA的正相关。我们的研究确定了肿瘤内Tregs的独特特征，并表明靶向Treg锁定转录因子，尤其是EOS，可能在基于Treg的抗肿瘤治疗中具有临床重要性。
