Screening cross-injury responses in skin organoids identifies a THBS2-enriched pro-repair fibroblast state during regeneration.

INTRODUCTION

Skin injuries impose a substantial global health burden. Conventional wound healing models fail to distinguish evolutionarily conserved regenerative mechanisms from injury specific responses because they cannot capture microenvironmental complexity.

OBJECTIVES

This study established a high-fidelity mouse skin organoid platform to simulate clinically relevant stressors and identify the core regulatory components of the regenerative program.

METHODS

Mature skin organoids were individually exposed to hypoxia, hyperthermia, or hyperinflammation. Transcriptomic analysis identified both divergent and convergent responses across diverse injury types. Single-cell RNA sequencing (scRNA-seq) was utilized to characterize fibroblast states in physiological and diabetic wounds.

RESULTS

A conserved 85-gene injury-responsive module was identified across all stressors, with thrombospondin 2 (Thbs2) emerging as a highly connected candidate hub within the extracellular matrix (ECM)-remodeling network. Integration with scRNA-seq datasets further showed that Thbs2 expression was enriched in an independently defined pro-repair fibroblast state characterized by ECM remodeling and repair-associated transcriptional features. In the diabetic wound microenvironment, this Thbs2-associated repair signature was attenuated, accompanied by accumulation of intermediate-state fibroblasts.

CONCLUSION

These findings highlight that the acquisition of a specific functional cellular state is closely linked to regeneration. This multi stress platform facilitates the development of universal pro-healing strategies by shifting the focus from molecular targeting to restoring functional cellular states.