Imaging-guided tape stripping reveals consistent stratum corneum profiling of disease severity and treatment-responsive molecular features in pediatric atopic dermatitis

Background: Atopic dermatitis (AD) is a common pediatric skin disorder with early onset and complex pathophysiology. Tape-strip sampling has enabled minimally invasive molecular profiling of skin lesions in children, yet approaches that verify and standardize stratum corneum (SC) sampling depth have remained limited. Objective: To evaluate the utility of imaging-guided tape stripping of SC, combined with transcriptomic and cytokine profiling, for characterizing molecular changes associated with disease severity and treatment response in pediatric AD. Methods: By coupling real-time imaging-guided tape stripping with RNA sequencing and cytokine profiling, we examined SC molecular signatures in healthy children (n = 15) and lesional and non-lesional skin of pediatric AD patients (n = 39) with heterogeneous disease severity, both at baseline and following 2 weeks of topical corticosteroid treatment. Results: Using imaging-verified SC sampling, we identified robust molecular signatures, including > 1100 differentially expressed genes (DEGs) between healthy and AD skin, and 144 DEGs between lesional and non-lesional AD skin (fold-change ≥2, FDR <0.1). Lesional skin showed enrichment of type 17- and type 2-associated immune pathways, tissue remodeling, and cell cycle control. Thirteen transcripts were significantly associated with clinical severity or itch, including CXCL8/IL8, CD300A, and FPR1. Most of these transcripts decreased after corticosteroid treatment, whereas CD300A and FCGR2B/CD32 remained elevated in a subset of children. Conclusion: These findings support the use of real-time imaging-guided tape stripping as a standardized, minimally invasive approach for verifying SC sampling depth and monitoring molecular changes associated with disease activity and treatment response in pediatric AD.