Department of Pulmonary and Infectious Diseases, Hannover Medical School, BREATH German Center for Lung Research (DZL), Feodor-Lynen-Str. 23, 30625, Hannover, Germany.
Department of Molecular Genetics, Institute of Health Carlos III, Institute for Rare Diseases Research, CIBER of Rare Diseases (CIBERER), Majadahonda, 28220, Madrid, Spain.
Lung. 2024 Apr;202(2):157-170. doi: 10.1007/s00408-024-00679-1. Epub 2024 Mar 17.
To investigate the transcriptome of human bronchial epithelial cells (HBEC) in response to serum from patients with different degrees of inflammation.
Serum from 19 COVID-19 patients obtained from the Hannover Unified Biobank was used. At the time of sampling, 5 patients had a WHO Clinical Progression Scale (WHO-CPS) score of 9 (severe illness). The remaining 14 patients had a WHO-CPS of below 9 (range 1-7), and lower illness. Multiplex immunoassay was used to assess serum inflammatory markers. The culture medium of HBEC was supplemented with 2% of the patient's serum, and the cells were cultured at 37 °C, 5% CO for 18 h. Subsequently, cellular RNA was used for RNA-Seq.
Patients with scores below 9 had significantly lower albumin and serum levels of E-selectin, IL-8, and MCP-1 than patients with scores of 9. Principal component analysis based on 500 "core genes" of RNA-seq segregated cells into two subsets: exposed to serum from 4 (I) and 15 (II) patients. Cells from a subset (I) treated with serum from 4 patients with a score of 9 showed 5566 differentially expressed genes of which 2793 were up- and 2773 downregulated in comparison with cells of subset II treated with serum from 14 patients with scores between 1 and 7 and one with score = 9. In subset I cells, a higher expression of TLR4 and CXCL8 but a lower CDH1, ACE2, and HMOX1, and greater effects on genes involved in metabolic regulation, cytoskeletal organization, and kinase activity pathways were observed.
This simple model could be useful to characterize patient serum and epithelial cell properties.
研究人支气管上皮细胞（HBEC）对不同炎症程度患者血清的转录组反应。
使用来自汉诺威统一生物库的 19 名 COVID-19 患者的血清。在取样时，5 名患者的世界卫生组织临床进展量表（WHO-CPS）评分为 9 分（重症）。其余 14 名患者的 WHO-CPS 评分低于 9 分（范围 1-7），病情较轻。采用多重免疫分析法评估血清炎症标志物。将 HBEC 的培养基中添加 2%的患者血清，并在 37°C、5%CO 下培养 18 小时。随后，使用细胞 RNA 进行 RNA-Seq。
评分低于 9 分的患者的白蛋白和血清 E-选择素、IL-8 和 MCP-1 水平明显低于评分 9 分的患者。基于 RNA-seq 的 500 个“核心基因”的主成分分析将细胞分为两个亚群：暴露于 4 名（I 组）和 15 名（II 组）患者血清中。与 II 组（用来自 4 名评分 9 分患者的血清处理的细胞）相比，来自亚群 I（用来自 14 名评分在 1 至 7 分之间和一名评分 9 分患者的血清处理的细胞）中处理的细胞有 5566 个差异表达基因，其中 2793 个上调，2773 个下调。在亚群 I 细胞中，TLR4 和 CXCL8 的表达更高，但 CDH1、ACE2 和 HMOX1 的表达更低，并且对代谢调节、细胞骨架组织和激酶活性途径中基因的影响更大。
该简单模型可用于表征患者血清和上皮细胞特性。
