Muraro E, Vaccher E, Furlan C, Fratta E, Fanetti G, Fae' D A, Martorelli D, Cangemi M, Polesel J, Navarria F, Gobitti C, Comaro E, Scaini C, Pratesi C, Zanussi S, Lupato V, Grando G, Giacomarra V, Sulfaro S, Barzan L, Dolcetti R, Steffan A, Canzonieri V, Franchin G
Immunopathology and Cancer Biomarkers Unit, Department of Translational Research, Centro di Riferimento Oncologico di Aviano (CRO) IRCCS, Aviano, PN, Italy.
Department of Medical Oncology, Centro di Riferimento Oncologico di Aviano (CRO) IRCCS, Aviano, PN, Italy.
Pathol Oncol Res. 2020 Oct;26(4):2459-2467. doi: 10.1007/s12253-020-00859-3. Epub 2020 Jun 21.
Undifferentiated Nasopharyngeal Carcinoma (UNPC) is associated with Epstein-Barr Virus (EBV) and characterized by an abundant immune infiltrate potentially influencing the prognosis. Thus, we retrospectively assessed the significance of immunosuppression in the UNPC microenvironment as prognostic biomarker of treatment failure in a non-endemic area, and monitored the variation of systemic EBV-specific immunity before and after chemoradiotherapy (CRT). DNA and RNA were extracted from diagnostic biopsies obtained by tumor and adjacent mucosa from 63 consecutive EBV+ UNPC patients who underwent radical CRT. Among these patients 11 relapsed within 2 years. The expression of the EBV-derived UNPC-specific BARF1 gene and several immune-related genes was monitored through quantitative RT-PCR and methylation-specific PCR analyses. Peripheral T cell responses against EBV and BARF1 were measured in 14 patients (7 relapses) through IFN-γ ELISPOT assay. We found significantly higher expression levels of BARF1, CD8, IFN-γ, IDO, PD-L1, and PD-1 in UNPC samples compared to healthy tissues. CD8 expression was significantly reduced in both tumor and healthy tissues in UNPC patients who relapsed within two years. We observed a hypomethylated FOXP3 intron 1 exclusively in relapsed UNPC patients. Finally, we noticed a significant decrease in EBV- and BARF1-specific T-cells after CRT only in relapsing patients. Our data suggest that a high level of immunosuppression (low CD8, hypomethylated FoxP3) in UNPC microenvironment may predict treatment failure and may allow an early identification of patients who could benefit from the addition of immune modulating strategies to improve first line CRT.
未分化鼻咽癌（UNPC）与爱泼斯坦-巴尔病毒（EBV）相关，其特征是存在丰富的免疫浸润，这可能会影响预后。因此，我们回顾性评估了在非流行地区，免疫抑制在UNPC微环境中作为治疗失败预后生物标志物的意义，并监测了放化疗（CRT）前后全身EBV特异性免疫的变化。从63例接受根治性CRT的连续EBV阳性UNPC患者的肿瘤及相邻黏膜的诊断活检组织中提取DNA和RNA。这些患者中有11例在2年内复发。通过定量RT-PCR和甲基化特异性PCR分析监测EBV衍生的UNPC特异性BARF1基因和几个免疫相关基因的表达。通过IFN-γ ELISPOT试验检测了14例患者（7例复发）外周血T细胞对EBV和BARF1的反应。我们发现，与健康组织相比，UNPC样本中BARF1、CD8、IFN-γ、吲哚胺2,3-双加氧酶（IDO）、程序性死亡受体配体1（PD-L1）和程序性死亡蛋白1（PD-1）的表达水平显著更高。在两年内复发的UNPC患者的肿瘤组织和健康组织中，CD8表达均显著降低。我们仅在复发的UNPC患者中观察到叉头框蛋白P3（FOXP3）内含子1低甲基化。最后，我们注意到仅在复发患者中，CRT后EBV和BARF1特异性T细胞显著减少。我们的数据表明，UNPC微环境中高水平的免疫抑制（低CD8、FOXP3低甲基化）可能预示治疗失败，并可能有助于早期识别那些可能从添加免疫调节策略中获益以改善一线CRT的患者。
