中性粒细胞胞外陷阱（NET）被归类为中性粒细胞炎症细胞死亡模式（NETosis）的一种形式，参与自然免疫和适应性免疫的调节。越来越多的研究揭示了NETs通过限制抗肿瘤效应细胞来重塑肿瘤免疫微环境（TIME）的能力，这可能会损害免疫治疗的疗效。为了探讨 NETs 相关基因是否对结肠癌 (COAD) 产生重要影响，我们进行了系统分析，并在本工作中展示了一些发现。首先，我们从癌症基因组图谱 (TCGA) 和基因表达综合 (GEO) 数据集中获取患者数据，旨在通过共识聚类检测两种与 NET 相关的亚型。为了注释NETs相关通路的作用，采用了基因本体富集分析。接下来，我们使用最小绝对收缩和选择算子 (LASSO) Cox 回归模型构建了 6 个新颖的 NETs 相关基因评分。我们发现 COAD 患者样本中 NETs 风险评分显着上调，且其水平与肿瘤临床病理和免疫特征显着相关。然后，根据NETs相关的分子亚型和风险特征，本研究比较了通过估计、CIBERSORT、TIMER、ssGSEA算法、肿瘤免疫功能障碍以及排除（TIDE）计算出的免疫细胞浸润。此外，我们证实 COAD 患者样本中 MPO（髓过氧化物酶）显着上调，其水平与肿瘤恶性程度和临床结果显着相关。此外，多重免疫组织化学(mIHC)空间分析证实MPO在空间位置上与Treg和PD-1 Treg密切相关，这表明MPO可能在TIME形成中发挥重要作用。总而言之，获得的结果表明六个 NETs 相关基因的预后特征有利于评估 COAD 患者的预后和化疗/免疫治疗的反应。© 2023。作者。

Neutrophil extracellular traps (NETs) have been categorized as a form of inflammatory cell death mode of neutrophils (NETosis) involved in natural immunity and the regulation of adaptive immunity. More and more studies revealed the ability of NETs to reshape the tumor immune microenvironment (TIME) by limiting antitumor effector cells, which may impair the efficacy of immunotherapy. To explore whether NETs-related genes make vital impacts on Colon carcinoma (COAD), we have carried out a systematic analysis and showed several findings in the present work. First, we obtained the patient's data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) dataset, aiming to detect two NETs-associated subtypes by consensus clustering. For the purpose of annotating the roles of NETs-related pathways, gene ontology enrichment analyses were adopted. Next, we constructed a 6 novel NETs-related genes score using the Least Absolute Shrinkage and Selection Operator (LASSO) Cox regression model. We found that the NETs risk score was notably upregulated in COAD patient samples, and its levels were notably correlated with tumor clinicopathological and immune traits. Then, according to NETs-associated molecular subtypes and the risk signature, this study compared immune cell infiltration calculated through the estimate, CIBERSORT, TIMER, ssGSEA algorithms, tumor immune dysfunction, as well as exclusion (TIDE). Furthermore, we confirm that MPO(myeloperoxidase) was significantly upregulated in COAD patient samples, and its levels were significantly linked to tumor malignancy and clinic outcome. Moreover, multiplex immunohistochemistry (mIHC) spatial analysis confirmed that MPO was closely related to Treg and PD-1 + Treg in spatial location which suggested MPO may paly an important role in TIME formation. Altogether, the obtained results indicated that a six NETs-related genes prognostic signature was conducive to estimating the prognosis and response of chemo-/immuno-therapy of COAD patients.© 2023. The Author(s).