鉴定和验证 CRLF1 和 NRG1 作为增生性瘢痕中的免疫相关标志物。
Identification and validation of CRLF1 and NRG1 as immune-related signatures in hypertrophic scar.
机构信息
Department of Plastic and Reconstructive Surgery, The Fourth Medical Center of Chinese PLA General Hospital, Beijing 100048, China; Chinese PLA Medical School, Beijing 100853, China.
Department of Urology, Peking University Third Hospital, Beijing 100191, China.
出版信息
Genomics. 2024 Mar;116(2):110797. doi: 10.1016/j.ygeno.2024.110797. Epub 2024 Jan 21.
BACKGROUND
Hypertrophic scar (HTS) is a prevalent chronic inflammatory skin disorder characterized by abnormal proliferation and extracellular matrix deposition and the precise mechanisms underlying HTS remain elusive. This study aimed to identify and validate potential immune-related genes associated with hypertrophic scar formation.
METHODS
Skin samples from normal (n = 12) and hypertrophic scar tissues (n = 12) were subjected to RNA-seq analysis. Differentially expressed genes (DEGs) and significant modular genes in Weighted gene Co-expression Network Analysis (WGCNA) were identified. Subsequently, functional enrichment analysis was performed on the intersecting genes. Additionally, eight immune-related genes were matched from the ImmPort database. Validation of NRG1 and CRLF1 was carried out using an external cohort (GSE136906). Furthermore, the association between these two genes and immune cells was assessed by Spearman correlation analysis. Finally, RNA was extracted from normal and hypertrophic scar samples, and RT-qPCR, Immunohistochemistry staining and Western Blot were employed to validate the expression of characteristic genes.
RESULTS
A total of 940 DEGs were identified between HTS and normal samples, and 288 key module genes were uncovered via WGCNA. Enrichment analysis in key module revealed involvement in many immune-related pathways, such as Th17 cell differentiation, antigen processing and presentation and B cell receptor signaling pathway. The eight immune-related genes (IFI30, NR2F2, NRG1, ESM1, NFATC2, CRLF1, COLEC12 and IL6) were identified by matching from the ImmPort database. Notably, we observed that activated mast cell positively correlated with CRLF1 expression, while CD8 T cells exhibited a positive correlation with NRG1. The expression of NRG1 and CRLF1 was further validated in clinical samples.
CONCLUSION
In this study, two key immune-related genes (CRLF1 and NRG1) were identified as characteristic genes associated with HTS. These findings provide valuable insights into the immune-related mechanisms underlying hypertrophic scar formation.
背景
增生性瘢痕（HTS）是一种常见的慢性炎症性皮肤疾病，其特征为异常增殖和细胞外基质沉积，但其确切发病机制仍不清楚。本研究旨在鉴定和验证与增生性瘢痕形成相关的潜在免疫相关基因。
方法
对 12 例正常皮肤和 12 例增生性瘢痕组织的皮肤样本进行 RNA-seq 分析。通过加权基因共表达网络分析（WGCNA）鉴定差异表达基因（DEGs）和显著模块基因。随后对交集基因进行功能富集分析。此外，从 ImmPort 数据库中匹配了 8 个免疫相关基因。使用外部队列（GSE136906）验证 NRG1 和 CRLF1 的表达。进一步通过 Spearman 相关分析评估这两个基因与免疫细胞的相关性。最后，从正常和增生性瘢痕样本中提取 RNA，采用 RT-qPCR、免疫组化染色和 Western blot 验证特征基因的表达。
结果
在 HTS 与正常样本之间共鉴定出 940 个 DEGs，通过 WGCNA 发现了 288 个关键模块基因。关键模块的富集分析显示，这些基因参与了许多免疫相关途径，如 Th17 细胞分化、抗原加工和呈递以及 B 细胞受体信号通路。通过从 ImmPort 数据库中匹配，鉴定出 8 个免疫相关基因（IFI30、NR2F2、NRG1、ESM1、NFATC2、CRLF1、COLEC12 和 IL6）。值得注意的是，我们观察到激活的肥大细胞与 CRLF1 的表达呈正相关，而 CD8 T 细胞与 NRG1 的表达呈正相关。NRG1 和 CRLF1 的表达在临床样本中进一步得到验证。
结论
在这项研究中，鉴定出两个关键的免疫相关基因（CRLF1 和 NRG1）作为与 HTS 相关的特征基因。这些发现为增生性瘢痕形成的免疫相关机制提供了有价值的见解。
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