Tape-Strip Proteomic Profiling of Atopic Dermatitis on Dupilumab Identifies Minimally Invasive Biomarkers

Abstract

Tape-stripping is a minimally invasive approach for skin sampling that captures the cutaneous immune/barrier abnormalities in atopic dermatitis (AD). However, tape-strips have not been used to evaluate molecular changes with therapeutic targeting. In this study, we sought to characterize the proteomic signature of tape-strips from AD patients, before and after dupilumab therapy. Twenty-six AD patients were treated with every-other-week dupilumab 300 mg for 16 weeks. Tape-strips from lesional and non-lesional skin were collected before and after treatment, and analyzed with the Olink proteomic assay. Using criteria of fold-change>1.5 and FDR < 0.05, 136 proteins significantly decreased after dupilumab treatment, corresponding to an overall mean improvement of 66.2% in the lesional vs. non-lesional AD proteome. Significant decreases after dupilumab were observed in immune markers related to general inflammation (MMP12), Th2 (CCL13/CCL17), Th17/Th22 (IL-12B, CXCL1, S100A12), and innate immunity (IL-6, IL-8, IL-17C), while the Th1 chemokines CXCL9/CXCL10 remained elevated. Proteins related to atherosclerosis/cardiovascular risk (e.g., SELE/E-selectin, IGFBP7, CHIT1/ chitotriosidase-1, AXL) also significantly decreased after treatment. Dupilumab therapy suppressed AD-related immune biomarkers and atherosclerosis/cardiovascular risk proteins. Tape-strip proteomics may be useful for monitoring therapeutic response in real-life settings, clinical trials, and longitudinal studies for AD and beyond.

Keywords: atherosclerosis; atopic dermatitis; cardiovascular disease; dupilumab; olink; proteomics; tape-strips; targeted therapeutics.

Figure 1

Heatmap of immune and atherosclerosis/cardiovascular proteins. Heatmap of immune and atherosclerosis and cardiovascular signaling proteins that are differentially expressed in tape-stripped skin of atopic dermatitis patients before and after dupilumab therapy, using the criteria of fold-change >1.5 and false discovery rate <0.05. Proteins are ordered by unsupervised hierarchical clustering, as represented by dendrogram on the left. The table on the right shows fold-changes in lesional vs. non-lesional tape-strips at baseline (LS vs. NL), and post- vs. pre-treatment with dupilumab (Post-Rx vs. Pre-Rx) for both non-lesional and lesional skin. LS, Lesional; NL, Non-lesional; Rx, Treatment.***FDR < 0.001, **FDR < 0.01, *FDR < 0.05, +FDR < 0.1.

Figure 2

Gene-set variation analysis. Mean z-scores of the Th2 (A), Th17 (B), Th1 (C), and atherosclerosis/cardiovascular risk (D) pathways in atopic dermatitis lesional and non-lesional tape-strips before and after dupilumb therapy, expressed as a boxplot. Red dots indicate mean values. Asterisks indicate significance between respective groups. AD, Atopic dermatitis; GSVA, Gene-set variation analysis; Rx, Treatment. ***FDR < 0.001, **FDR < 0.01.

Figure 3

Correlation plots. Spearman correlation scatter plots [linear regression [blue line] with its confidence interval [gray area]] for percent improvement in EASI score before and after dupilumab vs. percent decrease in protein expression levels, as measured by Olink. Upper left corner with r, Spearman correlation coefficient and p, associated P-value. EASI, Eczema Area and Severity Index.