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Deciphering fibroblast and myeloid-cell states in keloids: an integrative analysis identifies ANKRD1 as a
Rongrong Xie1, Wanting Yao1, Peipei Chen2
1Department of Plastic Surgery, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, China.
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.
