PDF(Pubmed)
Abstract：
BACKGROUND: The advent of immunotherapy targeting immune checkpoints has conferred significant clinical advantages to patients with lung adenocarcinoma (LUAD); However, only a limited subset of patients exhibit responsiveness to this treatment. Consequently, there is an imperative need to stratify LUAD patients based on their response to immunotherapy and enhance the therapeutic efficacy of these treatments.
METHODS: The differentially co-expressed genes associated with CD8 + T cells were identified through weighted gene co-expression network analysis (WGCNA) and the Search Tool for the Retrieval of Interacting Genes (STRING) database. These gene signatures facilitated consensus clustering for TCGA-LUAD and GEO cohorts, categorizing them into distinct immune subtypes (C1, C2, C3, and C4). The Tumor Immune Dysfunction and Exclusion (TIDE) model and Immunophenoscore (IPS) analysis were employed to assess the immunotherapy response of these subtypes. Additionally, the impact of inhibitors targeting five hub genes on the interaction between CD8 + T cells and LUAD cells was evaluated using CCK8 and EDU assays. To ascertain the effects of these inhibitors on immune checkpoint genes and the cytotoxicity mediated by CD8 + T cells, flow cytometry, qPCR, and ELISA methods were utilized.
RESULTS: Among the identified immune subtypes, subtypes C1 and C3 were characterized by an abundance of immune components and enhanced immunogenicity. Notably, both C1 and C3 exhibited higher T cell dysfunction scores and elevated expression of immune checkpoint genes. Multi-cohort analysis of Lung Adenocarcinoma (LUAD) suggested that these subtypes might elicit superior responses to immunotherapy and chemotherapy. In vitro experiments involved co-culturing LUAD cells with CD8 + T cells and implementing the inhibition of five pivotal genes to assess their function. The inhibition of these genes mitigated the immunosuppression on CD8 + T cells, reduced the levels of PD1 and PD-L1, and promoted the secretion of IFN-γ and IL-2.
CONCLUSIONS: Collectively, this study delineated LUAD into four distinct subtypes and identified five hub genes correlated with CD8 + T cell activity. It lays the groundwork for refining personalized therapy and immunotherapy strategies for patients with LUAD.
METHODS: The differentially co-expressed genes associated with CD8 + T cells were identified through weighted gene co-expression network analysis (WGCNA) and the Search Tool for the Retrieval of Interacting Genes (STRING) database. These gene signatures facilitated consensus clustering for TCGA-LUAD and GEO cohorts, categorizing them into distinct immune subtypes (C1, C2, C3, and C4). The Tumor Immune Dysfunction and Exclusion (TIDE) model and Immunophenoscore (IPS) analysis were employed to assess the immunotherapy response of these subtypes. Additionally, the impact of inhibitors targeting five hub genes on the interaction between CD8 + T cells and LUAD cells was evaluated using CCK8 and EDU assays. To ascertain the effects of these inhibitors on immune checkpoint genes and the cytotoxicity mediated by CD8 + T cells, flow cytometry, qPCR, and ELISA methods were utilized.
RESULTS: Among the identified immune subtypes, subtypes C1 and C3 were characterized by an abundance of immune components and enhanced immunogenicity. Notably, both C1 and C3 exhibited higher T cell dysfunction scores and elevated expression of immune checkpoint genes. Multi-cohort analysis of Lung Adenocarcinoma (LUAD) suggested that these subtypes might elicit superior responses to immunotherapy and chemotherapy. In vitro experiments involved co-culturing LUAD cells with CD8 + T cells and implementing the inhibition of five pivotal genes to assess their function. The inhibition of these genes mitigated the immunosuppression on CD8 + T cells, reduced the levels of PD1 and PD-L1, and promoted the secretion of IFN-γ and IL-2.
CONCLUSIONS: Collectively, this study delineated LUAD into four distinct subtypes and identified five hub genes correlated with CD8 + T cell activity. It lays the groundwork for refining personalized therapy and immunotherapy strategies for patients with LUAD.
摘要：
背景：针对免疫检查点的免疫疗法的出现为肺腺癌（LUAD）患者赋予了显着的临床优势；然而，只有有限的一部分患者对这种治疗有反应.因此,迫切需要根据LUAD患者对免疫治疗的反应对其进行分层,并提高这些治疗的疗效.
方法：通过加权基因共表达网络分析（WGCNA）和检索相互作用基因的搜索工具（STRING）数据库鉴定与CD8+T细胞相关的差异共表达基因。这些基因特征促进了TCGA-LUAD和GEO队列的共识聚类，将它们分为不同的免疫亚型(C1、C2、C3和C4)。采用肿瘤免疫功能障碍和排斥（TIDE）模型和免疫表型（IPS）分析来评估这些亚型的免疫疗法反应。此外,使用CCK8和EDU分析评估了靶向5种hub基因的抑制剂对CD8+T细胞和LUAD细胞之间相互作用的影响.为了确定这些抑制剂对免疫检查点基因和CD8+T细胞介导的细胞毒性的影响，流式细胞术,qPCR,采用ELISA方法。
结果：在确定的免疫亚型中，C1和C3亚型的特征在于丰富的免疫成分和增强的免疫原性.值得注意的是,C1和C3均表现出更高的T细胞功能障碍评分和免疫检查点基因表达升高.肺腺癌（LUAD）的多队列分析表明，这些亚型可能引起免疫疗法和化学疗法的良好反应。体外实验涉及将LUAD细胞与CD8+T细胞共培养并实施5个关键基因的抑制以评估其功能。这些基因的抑制减轻了对CD8+T细胞的免疫抑制,降低PD1和PD-L1的水平,促进IFN-γ和IL-2的分泌。
结论：总的来说，本研究将LUAD分为4种不同的亚型,并鉴定出5种与CD8+T细胞活性相关的hub基因.这为完善LUAD患者的个性化治疗和免疫治疗策略奠定了基础。
方法：通过加权基因共表达网络分析（WGCNA）和检索相互作用基因的搜索工具（STRING）数据库鉴定与CD8+T细胞相关的差异共表达基因。这些基因特征促进了TCGA-LUAD和GEO队列的共识聚类，将它们分为不同的免疫亚型(C1、C2、C3和C4)。采用肿瘤免疫功能障碍和排斥（TIDE）模型和免疫表型（IPS）分析来评估这些亚型的免疫疗法反应。此外,使用CCK8和EDU分析评估了靶向5种hub基因的抑制剂对CD8+T细胞和LUAD细胞之间相互作用的影响.为了确定这些抑制剂对免疫检查点基因和CD8+T细胞介导的细胞毒性的影响，流式细胞术,qPCR,采用ELISA方法。
结果：在确定的免疫亚型中，C1和C3亚型的特征在于丰富的免疫成分和增强的免疫原性.值得注意的是,C1和C3均表现出更高的T细胞功能障碍评分和免疫检查点基因表达升高.肺腺癌（LUAD）的多队列分析表明，这些亚型可能引起免疫疗法和化学疗法的良好反应。体外实验涉及将LUAD细胞与CD8+T细胞共培养并实施5个关键基因的抑制以评估其功能。这些基因的抑制减轻了对CD8+T细胞的免疫抑制,降低PD1和PD-L1的水平,促进IFN-γ和IL-2的分泌。
结论：总的来说，本研究将LUAD分为4种不同的亚型,并鉴定出5种与CD8+T细胞活性相关的hub基因.这为完善LUAD患者的个性化治疗和免疫治疗策略奠定了基础。
