Deciphering fibroblast and myeloid-cell states in keloids: an integrative analysis identifies ANKRD1 as a ferroptosis-related candidate gene.

BACKGROUND

Keloids are fibroproliferative skin disorders characterized by persistent extracellular matrix deposition and limited treatment options. Ferroptosis has been implicated in fibrosis, but the molecular basis of ferroptosis-related changes in keloids remains unclear.

METHODS

Fibroblast-derived bulk transcriptomic datasets (GSE44270 and GSE145725) were integrated to identify differentially expressed ferroptosis-related genes. Weighted gene co-expression network analysis and machine-learning algorithms were used to identify candidate hub genes, which were evaluated in an external fibroblast dataset (GSE7890). Whole-tissue single-cell RNA-sequencing data (GSE191067) were used to characterize fibroblast and myeloid-cell states in keloid and normal-skin samples. Candidate regulatory relationships and potential ANKRD1-binding compounds were assessed in silico. Clinical tissue specimens were used for histological and protein-expression validation.

RESULTS

A total of 163 differentially expressed genes were identified, and integration with 1,318 ferroptosis-related genes prioritized ANKRD1 as a candidate hub gene. ANKRD1 showed an area under the receiver operating characteristic curve above 0.80 in the external validation dataset and was positively associated with YAP1 expression in fibroblast-derived bulk transcriptomic data. Single-cell analysis identified a ECM-remodeling fibroblast state and three myeloid-cell states: inflammatory macrophages, resident-phagocytic macrophages, and DC-like myeloid cells. The ceRNA analysis identified a putative NEAT1/MALAT1-miR-493-5p-ANKRD1 regulatory axis. Molecular docking, molecular-dynamics simulation, and ADMET profiling prioritized oleanolic acid and quercetin as candidate ANKRD1-binding compounds. Histological analyses showed excessive collagen deposition and increased ANKRD1 protein expression in keloid tissues.

CONCLUSION

ANKRD1 is a ferroptosis-related, fibroblast-associated candidate gene in keloids. The single-cell data describe fibroblast and myeloid-cell states in the keloid microenvironment. The proposed regulatory axis, candidate compounds, and potential role of ANKRD1 in fibroblast-myeloid interactions require further spatial and functional validation.