Multi-Omic Integration of Single-Cell, Spatial, and Bulk RNA-Seq Deciphers Endothelial Heterogeneity and WWTR1-Mediated Vascular Reprogramming in Keloids

Keloids are complex fibroproliferative disorders characterized by persistent inflammation and vascular dysfunction. However, the high-resolution cellular landscape of their endothelial niche remains poorly defined due to the limited availability of specialized transcriptomic datasets. In this study, we addressed this critical knowledge gap by integrating single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics with our in-house bulk RNA-seq data from clinical keloid and normal skin samples, thereby significantly expanding the available transcriptomic resources for keloid vascular research. Our multi-omic analysis identified ten distinct endothelial cell (EC) subpopulations and revealed a marked expansion of a pro-pathological “Stress-endo” cluster within the keloid microenvironment. By leveraging our self-sequenced clinical data for cross-validation, we characterized the Stress-endo subpopulation as a pivotal mechanical-inflammatory nexus and identified WWTR1 (TAZ) as the master transcription factor driving its pathological transition. Developmental trajectory and cell–cell communication analyses further demonstrated that this activated EC niche actively orchestrates immune cell recruitment and tissue remodeling through IL-17 and Visfatin signaling pathways. The upregulation of WWTR1 in keloid tissues was further validated at both the mRNA and protein levels by quantitative PCR and western blotting, respectively. Collectively, our study provides an enriched and refined transcriptomic framework for keloid pathogenesis, positioning WWTR1-mediated endothelial stress as a central orchestrator of chronic inflammation and fibrosis, thereby offering a more robust foundation for future vascular-targeted therapeutic strategies.