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Single-cell transcriptomics combined with interstitial fluid proteomics defines cell type-specific immune regulation
Thomas B Rojahn1, Vera Vorstandlechner2, Thomas Krausgruber3
1Department of Dermatology, Medical University of Vienna, Vienna, Austria.
The Journal of Allergy and Clinical Immunology
|April 29, 2020
Summary
Suction blistering effectively captures skin cells and interstitial fluid for atopic dermatitis (AD) research. This method offers superior transcriptomic resolution and reveals novel myeloid cell involvement in AD pathogenesis.
Area of Science:
- Dermatology
- Immunology
- Genomics
- Proteomics
Background:
- Atopic dermatitis (AD) is a prevalent chronic skin disease with incompletely understood pathogenesis.
- Limited treatment options exist due to the complex nature of AD.
- Characterizing AD at molecular levels is crucial for therapeutic advancements.
Purpose of the Study:
- To comprehensively characterize atopic dermatitis (AD) using transcriptomic and proteomic analyses in humans.
- To evaluate the efficacy of skin suction blistering as a method for sampling AD tissues.
- To compare molecular profiles obtained from suction blistering with conventional skin biopsies.
Main Methods:
- Skin suction blistering was employed as a minimally invasive technique to collect epidermal cells and interstitial fluid.
- Single-cell RNA sequencing was utilized to analyze transcriptional profiles of cells from blister and biopsy samples.
- Proteomic analysis of blister fluid was performed to identify key proteins associated with AD.
Main Results:
- Suction blistering captured epidermal and immune cells effectively, comparable to biopsies, with enhanced resolution for certain transcripts.
- Transcriptional analysis revealed characteristic cytokine upregulation (IL13, IL22) in specific T cell subsets and identified myeloid cells as the most enriched cell type in AD.
- Proteomic analysis of blister fluid highlighted upregulated dendritic cell and macrophage products, indicating their role in AD.
- Mast cells and CD163+ macrophages were less represented in blister samples compared to biopsies.
Conclusions:
- Skin suction blistering is a valuable tool for AD research, offering advantages over conventional biopsies.
- This method provides superior transcriptomic resolution and integrates proteomic data from interstitial fluid.
- Novel inflammatory players and cellular mechanisms in the AD microenvironment were uncovered, advancing our understanding of the disease.
