Unraveling the biological potential of skin fibroblast

Abstract

Objective: In this study, we examined the molecular response of human skin fibroblasts to an inflammatory cytokine to evaluate their suitability as models for immunopathology research. Methods: Skin fibroblasts were stimulated with tumour necrosis factor (TNF)-α, and the transcriptome was profiled via RNA-Seq. The differentially expressed genes were screened to predict immunological pathways and interactions. The cytokines and signaling pathways were validated at protein level. Similarly to immune cells, TNF-α caused transcriptional and transductional changes in fibroblasts. Results: Functional analysis revealed significant enrichment of TNF-α signaling and cell chemotaxis (normalized enrichment score = 2.59 and 3.42). We also detected enrichment of nuclear factor kappa B (NF-κB) target genes and NF-kB activation, confirmed by complete protein degradation of its inhibitor IκBa (p = 0.0019). The MAPK/ERK and p38 MAPK pathways were also activated. Finally, we observed significant secretion of proinflammatory cytokines and chemokines, such as interleukin 6 (p = 0.02), CXCL8 (p = 0.027), CCL2 (p = 0.028) and CCL5 (p = 0.016). Conclusion: This study advances the biological understanding of skin fibroblast responses to TNF-α, revealing their intracellular pathways and secretome. It discloses techniques for leveraging fibroblasts' potential as in vitro models to identify inflammatory drivers, particularly when alternative models are inaccessible.