Identification and Preliminary mRNA Validation of Key Genes Associated with Tolerogenic Dendritic Cells in Keloid Based on Transcriptome Data.

PURPOSE: Keloid is a cutaneous fibrotic disorder characterized by excessive fibrous tissue proliferation during wound healing. Tolerogenic dendritic cells (tolDCs) may participate in keloid immunopathology, but the related molecular signatures remain unclear. This study aimed to identify candidate tolDC-associated genes in keloid by integrating tolDC-related transcriptomic profiles with keloid transcriptomic datasets.

PATIENTS AND

METHODS

Public keloid and tolDC transcriptome datasets were analyzed. Candidate genes were screened by intersecting keloid differentially expressed genes (DEGs) with tolDC-related DEGs. Key genes were selected using protein-protein interaction analysis, machine learning algorithms, expression comparison, and ROC analysis. An ANN model based on the selected genes was constructed and validated. Enrichment analysis, subcellular localization, immune infiltration analysis, regulatory network construction, drug prediction, and molecular docking were performed to explore their potential biological relevance. The expression of key genes was preliminarily validated at the mRNA level in clinical samples by RT-qPCR.

RESULTS

A total of 2,182 keloid DEGs and 3,541 tolDC-related genes were intersected, and CCL20 and NLRC4 were identified as candidate key genes. The ANN model showed potential diagnostic value for distinguishing keloid from control samples. CCL20 and NLRC4 were mainly localized in the cytoplasm and were associated with pathways including Reactome antimicrobial peptide signaling. Immune infiltration analysis showed associations with six immune cell types, including eosinophils and memory B cells. Regulatory network analysis identified 31 miRNAs potentially targeting these genes, and drug prediction suggested associations with 79 chemical compounds. RT-qPCR analysis in clinical samples supported the bioinformatics findings at the mRNA level.

CONCLUSION

This study identified CCL20 and NLRC4 as candidate tolDC-associated genes in keloid. These genes may provide candidate diagnostic biomarkers and hypotheses for future immunomodulatory therapeutic targets in keloid.