DataSheet1_Characterization of Necroptosis-Related Molecular Subtypes and Therapeutic Response in Lung Adenocarcinoma.xlsx

Lung adenocarcinoma (LUAD) is one of the most common malignant tumors with high morbidity and mortality and is usually associated with therapeutic resistance and poor prognosis because of individual biological heterogeneity. There is an unmet need to screen for reliable parameters, especially immunotherapy-related biomarkers to predict the patient’s outcomes. Necroptosis is a special caspase-independent form of necrotic cell death associated with the pathogenesis, progression, and prognosis of multiple tumors but the potential connection between necroptosis-related genes (NRGs) and LUAD still remains unclear. In this study, we expounded mutational and transcriptional alterations of 67 NRGs in 522 LUAD samples and proposed a consensus-clustering subtype of these patients into two cohorts with distinct immunological and clinical prognosis characteristics. Cluster B patients were associated with a better prognosis and characterized by relatively lower expression of NRGs, higher immune scores in the tumor microenvironment (TME), more mild clinical stages, and downregulated expression of immunotherapy checkpoints. Subsequently, the NRG score was further established to predict the overall survival (OS) of LUAD patients using univariate Cox, LASSO, and multivariate Cox regression analyses. The immunological characteristics and potential predictive capability of NRG scores were further validated by 583 LUAD patients in external datasets. In addition to better survival and immune-activated conditions, low-NRG-score cohorts exhibited a significant positive correlation with the mRNA stem index (mRNAsi) and tumor mutation burden (TMB) levels. Combined with classical clinical characteristics and NRG scores, we successfully defined a novel necroptosis-related nomogram to accurately predict the 1/3/5-year survival rate of individual LUAD patients, and the potential predictive capability was further estimated and validated in multiple test datasets with high AUC values. Integrated transcriptomic analysis helps us seek vital NRGs and supplements a novel clinical application of NRG scores in predicting the overall survival and therapeutic benefits for LUAD patients.

肺腺癌（LUAD）是最常见的恶性肿瘤之一，兼具高发病率与高死亡率，且因个体生物学异质性，常伴随治疗耐药与不良预后。目前临床仍存在未被满足的需求：亟需筛选可靠的预测参数，尤其是免疫治疗相关生物标志物，以预判患者转归。坏死性凋亡是一种特殊的不依赖半胱氨酸天冬氨酸蛋白酶（caspase）的坏死性细胞死亡形式，其与多种肿瘤的发病机制、疾病进展及预后密切相关，但坏死性凋亡相关基因（NRGs）与肺腺癌之间的潜在关联仍不明确。
本研究对522份肺腺癌样本中67个NRGs的突变与转录组改变进行了系统剖析，并基于共识聚类将患者分为两个队列，二者具备截然不同的免疫学特征与临床预后表型。队列B患者预后更佳，其特征为NRGs表达水平相对较低、肿瘤微环境（TME）免疫评分更高、临床分期相对偏早，且免疫检查点表达下调。
随后，本研究通过单变量Cox回归、最小绝对收缩与选择算子（LASSO）回归及多变量Cox回归分析，构建了NRG评分模型以预测肺腺癌患者的总生存期（OS）。借助包含583份肺腺癌样本的外部数据集，本研究进一步验证了NRG评分的免疫学特征与潜在预测效能。除具备更优的生存结局与免疫激活状态外，低NRG评分队列还与mRNA干细胞指数（mRNAsi）及肿瘤突变负荷（TMB）水平呈显著正相关。
本研究结合经典临床特征与NRG评分，成功构建了一种新型坏死性凋亡相关列线图，可精准预测单个肺腺癌患者的1/3/5年生存率，其潜在预测效能在多个测试数据集中得到了进一步评估与验证，曲线下面积（AUC）值较高。整合转录组学分析有助于我们筛选关键NRGs，并为NRG评分在预测肺腺癌患者总生存期与治疗获益方面提供了全新的临床应用方向。