小气道上皮的失巢凋亡抵抗参与慢性阻塞性肺疾病的进展。
Anoikis resistance of small airway epithelium is involved in the progression of chronic obstructive pulmonary disease.
机构信息
Department of Respiratory and Critical Care Medicine, Peking University Third Hospital, Beijing, China.
Department of Emergency Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
出版信息
Front Immunol. 2023 Apr 5;14:1155478. doi: 10.3389/fimmu.2023.1155478. eCollection 2023.
BACKGROUND
Anoikis resistance is recognized as a crucial step in the metastasis of cancer cells. Most epithelial tumors are distinguished by the ability of epithelial cells to abscond anoikis when detached from the extracellular matrix. However, no study has investigated the involvement of anoikis in the small airway epithelium (SAE) of chronic obstructive pulmonary disease (COPD).
METHODS
Anoikis-related genes (ANRGs) exhibiting differential expression in COPD were identified using microarray datasets obtained from the Gene Expression Omnibus (GEO) database. Unsupervised clustering was performed to classify COPD patients into anoikis-related subtypes. Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, gene set enrichment analysis (GSEA), and gene set variation analysis (GSVA) were used to annotate the functions between different subtypes. Differential expression analysis and weighted gene co-expression network analysis (WGCNA) were leveraged to identify key molecules. The relative proportion of infiltrating immune cells in the SAE was quantified using the CIBERSORT and ssGSEA computational algorithms, and the correlation between key molecules and immune cell abundance was analyzed. The expression of key molecules in BEAS-2B cells exposed to cigarette smoke extract (CSE) was validated using qRT-PCR.
RESULTS
A total of 25 ANRGs exhibited differential expression in the SAE of COPD patients, based on which two subtypes of COPD patients with distinct anoikis patterns were identified. COPD patients with anoikis resistance had more advanced GOLD stages and cigarette consumption. Functional annotations revealed a different immune status between COPD patients with pro-anoikis and anoikis resistance. Tenomodulin (TNMD) and long intergenic non-protein coding RNA 656 (LINC00656) were subsequently identified as key molecules involved in this process, and a close correlation between TNMD and the infiltrating immune cells was observed, such as activated CD4 memory T cells, M1 macrophages, and activated NK cells. Further enrichment analyses clarified the relationship between TNMD and the inflammatory and apoptotic signaling pathway as the potential mechanism for regulating anoikis. experiments showed a dramatic upregulation of TNMD and LINC00656 in BEAS-2B cells when exposed to 3% CSE for 48 hours.
CONCLUSION
TNMD contributes to the progression of COPD by inducing anoikis resistance in SAE, which is intimately associated with the immune microenvironment.
背景
抗失巢凋亡被认为是癌细胞转移的关键步骤。大多数上皮肿瘤的特点是上皮细胞从细胞外基质中脱离时能够逃避失巢凋亡。然而，目前尚无研究探讨失巢凋亡在慢性阻塞性肺疾病（COPD）的小气道上皮（SAE）中的作用。
方法
从基因表达综合数据库（GEO）数据库中获取的微阵列数据集，鉴定出 COPD 中差异表达的抗失巢凋亡相关基因（ANRGs）。采用无监督聚类方法将 COPD 患者分为抗失巢凋亡相关亚型。进行基因本体论（GO）分析、京都基因与基因组百科全书（KEGG）分析、基因集富集分析（GSEA）和基因集变异分析（GSVA），以注释不同亚型之间的功能。进行差异表达分析和加权基因共表达网络分析（WGCNA），以鉴定关键分子。利用 CIBERSORT 和 ssGSEA 计算算法定量分析 SAE 中浸润免疫细胞的相对比例，并分析关键分子与免疫细胞丰度的相关性。采用 qRT-PCR 验证香烟烟雾提取物（CSE）暴露的 BEAS-2B 细胞中关键分子的表达。
结果
基于 25 个在 COPD 患者 SAE 中差异表达的 ANRGs，鉴定出两种具有不同失巢凋亡模式的 COPD 患者亚型。具有抗失巢凋亡能力的 COPD 患者的 GOLD 分期和吸烟量更高。功能注释显示，具有促失巢凋亡和抗失巢凋亡特征的 COPD 患者的免疫状态不同。随后鉴定出腱调蛋白（TNMD）和长链非编码 RNA656（LINC00656）为该过程中的关键分子，并且观察到 TNMD 与浸润免疫细胞之间存在密切相关性，例如激活的 CD4 记忆 T 细胞、M1 巨噬细胞和激活的 NK 细胞。进一步的富集分析阐明了 TNMD 与炎症和凋亡信号通路之间的关系，这可能是调节失巢凋亡的潜在机制。实验表明，暴露于 3% CSE 48 小时后，BEAS-2B 细胞中的 TNMD 和 LINC00656 显著上调。
结论
TNMD 通过诱导 SAE 中的抗失巢凋亡作用，促进 COPD 的进展，这与免疫微环境密切相关。
Zotero 插件