一项与肺腺癌中二硫键过氧化相关的预后标志物的构建和验证，结合单细胞 RNA 测序和批量 RNA 测序技术。

BACKGROUND: Lung adenocarcinoma (LUAD), the predominant subtype of non-small cell lung cancer (NSCLC), remains a pervasive global public health concern. Disulfidoptosis, a nascent form of regulated cell death (RCD), presents an emerging field of inquiry. Currently, investigations into disulfidoptosis are in their initial stages. Our undertaking sought to integrate single-cell RNA sequencing (scRNA-seq) in conjunction with traditional bulk RNA sequencing (bulk RNA-seq) methodologies, with the objective of delineating genes associated with disulfidoptosis and subsequently prognosticating the clinical outcomes of LUAD patients. METHODS: Initially, we conducted an in-depth examination of the cellular composition disparities existing between LUAD and normal samples using scRNA-seq data sourced from GSE149655. Simultaneously, we scrutinized the expression patterns of disulfidoptosis-associated gene sets across diverse cell types. Subsequently, leveraging the bulk RNA-seq data, we formulated disulfidoptosis-related prognostic risk signatures (DRPS) employing LASSO-Cox regression. This was accomplished by focusing on genes implicated in disulfidoptosis that exhibited differential expression within endothelial cells (ECs). Sequentially, the robustness and precision of the DRPS model were rigorously verified through both internal and external validation datasets. In parallel, we executed single-cell trajectory analysis to delve into the differentiation dynamics of ECs. Concluding our study, we undertook a comprehensive investigation encompassing various facets. These included comparative assessments of enrichment pathways, clinicopathological parameters, immune cell abundance, immune response-associated genes, impacts of immunotherapy, and drug predictions among distinct risk cohorts. RESULTS: The scrutiny of scRNA-seq data underscored discernible disparities in cellular composition between LUAD and normal samples. Furthermore, disulfidoptosis-associated genes exhibited marked discrepancies within endothelial cells (ECs). Consequently, we formulated the Disulfidoptosis-Related Prognostic Signature (DRPS) to facilitate prognostic prediction. The prognostic nomogram based on the risk score effectively demonstrated DRPS's robust capacity to prognosticate survival outcomes. This assertion was corroborated by rigorous assessments utilizing both internal and external validation sets, thus affirming the commendable predictive accuracy and enduring stability of DRPS. Functional enrichment analysis shed light on the significant correlation of DRPS with pathways intrinsic to the cell cycle. Subsequent analysis unveiled correlations between DRPS and gene mutations characteristic of LUAD, as well as indications of an immunosuppressive status. Through drug prediction, we explored potential therapeutic agents for low-risk patients. Concluding our investigation, qRT-PCR experiments confirmed the heightened expression levels of EPHX1, LDHA, SHC1, MYO6, and TLE1 in lung cancer cell lines.

背景：肺腺癌（LUAD）是非小细胞肺癌（NSCLC）的主要亚型，仍然是一个普遍存在的全球公共卫生问题。二硫键过氧化（disulfidoptosis）是一种新兴的调控细胞死亡（RCD）形式，是一个新兴的研究领域。目前，对二硫键过氧化的研究还处于起步阶段。我们的研究旨在将单细胞 RNA 测序（scRNA-seq）与传统的批量 RNA 测序（bulk RNA-seq）方法相结合，以确定与二硫键过氧化相关的基因，并随后预测 LUAD 患者的临床结局。

方法：我们首先使用 GSE149655 中的 scRNA-seq 数据深入研究 LUAD 和正常样本之间的细胞组成差异。同时，我们研究了不同细胞类型中二硫键过氧化相关基因集的表达模式。随后，我们利用批量 RNA-seq 数据，通过 LASSO-Cox 回归构建了二硫键过氧化相关预后风险特征（DRPS）。这是通过关注在血管内皮细胞（ECs）中表现出差异表达的与二硫键过氧化相关的基因来实现的。随后，我们通过内部和外部验证数据集严格验证了 DRPS 模型的稳健性和准确性。同时，我们进行了单细胞轨迹分析，深入研究了 ECs 的分化动态。在研究结束时，我们进行了全面的研究，包括比较富集途径、临床病理参数、免疫细胞丰度、免疫反应相关基因、不同风险队列的免疫治疗影响和药物预测等方面。

结果：scRNA-seq 数据的分析表明，LUAD 和正常样本之间的细胞组成存在明显差异。此外，二硫键过氧化相关基因在血管内皮细胞（ECs）中表现出明显差异。因此，我们构建了二硫键过氧化相关预后签名（DRPS）以进行预后预测。基于风险评分的预后列线图有效地证明了 DRPS 对生存结局的预测能力。这一结论得到了内部和外部验证集的严格评估的支持，从而证实了 DRPS 具有良好的预测准确性和持久的稳定性。功能富集分析表明，DRPS 与细胞周期相关的途径有显著相关性。进一步的分析表明，DRPS 与 LUAD 特征性的基因突变以及免疫抑制状态相关。通过药物预测，我们探索了低风险患者的潜在治疗药物。在研究结束时，qRT-PCR 实验证实了 EPHX1、LDHA、SHC1、MYO6 和 TLE1 在肺癌细胞系中的高表达水平。