DataSheet_1_Intratumoral neutrophil extracellular traps are associated with unfavorable clinical outcomes and immunogenic context in pancreatic ductal adenocarcinoma.pdf

Extracellular traps (ETs) and tumor-infiltrating immune cells play crucial roles in tumor progression. However, little is known about the clinical significance of tumor-infiltrating neutrophils and macrophages and the related ETs in pancreatic ductal adenocarcinoma (PDAC). This study investigates the associations between neutrophil or macrophage infiltration or ET formation and the clinicopathological features, molecular characteristics, immune checkpoint molecules, clinical outcomes, and response to adjuvant chemotherapy (ACT) in PDAC. We performed multiplex immunofluorescence staining to detect ET formation by neutrophils or macrophages using tissue microarrays obtained from 205 patients, and analyzed the immunohistochemistry data for PD-L1, PD-L2, B7-H3, and B7-H4. The ET expression rates in macrophages and neutrophils were 23.9% and 45.4%, respectively. Patients with a high density of neutrophils or positive expression of neutrophil ETs exhibited poorer progression-free survival (PFS) and disease-specific survival (DSS), whereas macrophage ETs were not related to PFS and DSS. Neutrophil infiltration and ET formation were identified as independent prognostic predictors of DSS using univariate and multivariate Cox analyses. Patients with PDAC with lower neutrophil infiltration or negative staining for neutrophil ETs are more likely to benefit from ACT. Patients with PDAC were more accurately stratified based on the infiltration of neutrophils and presence of neutrophil ETs, and patients with low neutrophil infiltration and negative staining for neutrophil ETs showed the best survival. Patients with positive neutrophil ETs demonstrated inferior DSS compared to those with negative neutrophil ETs in the PD-L1 tumor proportion score (TPS) < 1% and PD-L1 IC < 1% subgroups. However, the positive expression of neutrophil ETs was not related to DSS in the PD-L1 TPS ≥ 1% or PD-L1 IC ≥ 1% subgroup. Our findings emphasize the potential of neutrophil infiltration and ETs as prognostic markers that could guide the formulation of more effective personalized treatments for PDAC.

细胞外陷阱（Extracellular traps, ETs）与肿瘤浸润免疫细胞在肿瘤进展中发挥关键作用。然而，目前对胰腺导管腺癌（pancreatic ductal adenocarcinoma, PDAC）中肿瘤浸润中性粒细胞、巨噬细胞及其相关ETs的临床意义尚不清楚。本研究探讨了PDAC中中性粒细胞或巨噬细胞浸润、ET形成与临床病理特征、分子特征、免疫检查点分子、临床结局及辅助化疗（adjuvant chemotherapy, ACT）应答之间的关联。我们采用来自205例患者的组织微阵列，通过多重免疫荧光染色检测中性粒细胞或巨噬细胞的ET形成情况，并对PD-L1、PD-L2、B7-H3及B7-H4的免疫组织化学数据进行分析。巨噬细胞与中性粒细胞的ET表达率分别为23.9%与45.4%。中性粒细胞密度较高或中性粒细胞ETs呈阳性表达的患者，其无进展生存期（progression-free survival, PFS）与疾病特异性生存期（disease-specific survival, DSS）更差；而巨噬细胞ETs与PFS、DSS无显著关联。通过单因素及多因素Cox分析，中性粒细胞浸润与ET形成被确定为DSS的独立预后预测因子。中性粒细胞浸润较低或中性粒细胞ETs染色呈阴性的PDAC患者，更有可能从辅助化疗中获益。结合中性粒细胞浸润情况与中性粒细胞ETs的存在情况，可对PDAC患者进行更精准的分层；其中，中性粒细胞浸润较低且中性粒细胞ETs染色呈阴性的患者生存情况最佳。在PD-L1肿瘤比例评分（tumor proportion score, TPS）<1%及PD-L1免疫细胞评分（immune cell score, IC）<1%的亚组中，中性粒细胞ETs呈阳性的患者DSS劣于中性粒细胞ETs呈阴性的患者。但在PD-L1 TPS≥1%或PD-L1 IC≥1%的亚组中，中性粒细胞ETs的阳性表达与DSS无显著关联。本研究结果凸显了中性粒细胞浸润与ETs作为预后标志物的潜力，可为PDAC制定更有效的个体化治疗方案提供指导。