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. Skin samples from normal ( = 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. 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. 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.

中文翻译：

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CRLF1 和 NRG1 作为增生性疤痕免疫相关特征的鉴定和验证

肥厚性疤痕 (HTS) 是一种常见的慢性炎症性皮肤病，其特征是异常增殖和细胞外基质沉积，但 HTS 的确切机制仍然难以捉摸。本研究旨在识别和验证与肥厚性疤痕形成相关的潜在免疫相关基因。对正常 (= 12) 和肥厚性疤痕组织 (n = 12) 的皮肤样本进行 RNA-seq 分析。在加权基因共表达网络分析（WGCNA）中鉴定出差异表达基因（DEG）和显着的模块基因。随后，对交叉基因进行功能富集分析。此外，还从 ImmPort 数据库中匹配了八个免疫相关基因。 NRG1 和 CRLF1 的验证是使用外部队列 (GSE136906) 进行的。此外，通过斯皮尔曼相关分析评估了这两个基因和免疫细胞之间的关联。最后，从正常和增生性疤痕样本中提取RNA，并采用RT-qPCR、免疫组化染色和Western Blot来验证特征基因的表达。通过WGCNA，HTS与正常样本之间共鉴定出940个DEG，并发现了288个关键模块基因。关键模块的富集分析揭示了其参与许多免疫相关通路，如Th17细胞分化、抗原加工和呈递以及B细胞受体信号通路。通过与ImmPort数据库匹配，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，我们观察到活化的肥大细胞与 CRLF1 表达呈正相关，而 CD8 T 细胞与 NRG1 表达呈正相关。 NRG1和CRLF1的表达在临床样本中得到进一步验证。在这项研究中，两个关键的免疫相关基因（CRLF1和NRG1）被确定为与HTS相关的特征基因。这些发现为了解增生性疤痕形成的免疫相关机制提供了宝贵的见解。