Single-cell transcriptomics combined with interstitial fluid proteomics defines cell type–specific immune regulation in atopic dermatitis
Journal of Allergy and Clinical Immunology, 2020
Rojahn T., Vorstandlechner V., Krausgruber T., Bauer W., Alkon N., Bangert C., Thaler F., Sadeghyar F., Fortelny N., Gernedl V., Rindler K., Elbe-Bürger A., Bock C., Mildner M., Brunner P.
| Disease area | Application area | Sample type | Products |
|---|---|---|---|
Immunological & Inflammatory Diseases Dermatological Diseases | Pathophysiology | Interstitial Fluid (skin blisters) | O Olink Target 96 |
Abstract
Background
Atopic dermatitis (AD) is the most common chronic inflammatory skin disease, but its complex pathogenesis is only insufficiently understood, resulting in still limited treatment options.
Objective
To characterize AD on both transcriptomic and proteomic levels in humans.
Methods
We used skin suction blistering, a painless and non-scarring procedure that can simultaneously sample skin cells and interstitial fluid, and compared results to conventional biopsies.
Results
Suction blistering captured epidermal and most immune cells equally well as biopsies, except for mast cells and non-migratory CD163+ macrophages that were only present in biopsy isolates. Using single-cell RNA sequencing, we found comparable transcriptional profiles of key inflammatory pathways between blister and biopsy AD, but suction blistering was superior in cell-specific resolution for high-abundance transcripts (KRT1/KRT10, KRT16/KRT6A, S100A8/S100A9), which showed some background signals in biopsy isolates. Compared to healthy controls, we found characteristic upregulation of AD-typical cytokines such as IL13 and IL22 in Th2 and Th22 cells, respectively, but we also discovered these mediators in proliferating T-cells and NKT-cells, that also expressed the antimicrobial cytokine IL26. Overall, not T-cells, but myeloid cells were most strongly enriched in AD, and we found dendritic cell (CLEC7A, amphiregulin/AREG, EREG) and macrophage products (CCL13) among the top-upregulated proteins in AD blister fluid proteomic analyses.
Conclusion
These data show that by using cutting-edge technology, suction blistering offers several advantages over conventional biopsies, including better transcriptomic resolution of skin cells, combined with proteomic information from interstitial fluid, unraveling novel inflammatory players that shape the cellular and proteomic microenvironment of AD.