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| Status |
Public on Jul 04, 2020 |
| Title |
Single-cell Transcriptomics Combined With Interstitial Fluid Proteomics Defines Cell Type-Specific Immune Regulation in Atopic Dermatitis |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
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: We sought to characterize AD on both transcriptomic and proteomic levels in humans.
Methods: We used skin suction blistering, a painless and nonscarring procedure that can simultaneously sample skin cells and interstitial fluid. We then compared results with conventional biopsies.
Results: Suction blistering captured epidermal and most immune cells equally well as biopsies, except for mast cells and nonmigratory 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 with 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 natural killer T 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.
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| Overall design |
Comparison of skin cells obtained by skin suction blistering and conventional biopsy
>>>Raw data are unvailable due to patient privacy concerns<<<
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| Contributor(s) |
Rojahn TB, Vorstandlechner V, Krausgruber T, Bauer WM, Alkon N, Bangert C, Thaler FM, Sadeghyar F, Fortelny N, Gernedl V, Rindler K, Elbe-Bürger A, Bock C, Mildner M, Brunner PM |
| Citation(s) |
32344053, 33717163 |
| Submission date |
Jul 03, 2020 |
| Last update date |
Mar 16, 2021 |
| Contact name |
Patrick M Brunner |
| E-mail(s) |
patrick.brunner@meduniwien.ac.at
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| Organization name |
Medical University of Vienna
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| Department |
Dermatology
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| Street address |
Waehringer Guertel 18-20
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| City |
Vienna |
| ZIP/Postal code |
1090 |
| Country |
Austria |
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| Platforms (1) |
| GPL21290 |
Illumina HiSeq 3000 (Homo sapiens) |
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| Samples (15)
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| Relations |
| BioProject |
PRJNA643988 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE153760_RAW.tar |
306.6 Mb |
(http)(custom) |
TAR (of MTX, TSV) |
| Raw data not provided for this record |
| Processed data provided as supplementary file |
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