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| Status |
Public on Jun 17, 2019 |
| Title |
Single-cell RNA-sequencing of skin, fresh blood, and cultured peripheral blood mononuclear cells from a patient with drug-induced hypersensitivity syndrome and healthy volunteers |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Purpose: Determine new therapeutic targets for a refractory drug-induced hypersensitivity syndrome/DRESS using single cell transcriptomic analysis.
Methods: Skin cells were dissociated from skin biopsies of a patient with DRESS and 5 healthy volunteers using enzymatic digestion, then viable skin cells were sorted using flow cytometry. Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood using Ficoll-paque density separation. PBMCs were cultured with or without medications for 4 days. In detail, 200,000 PBMCs were cultivated in 200ul of RPMI-1640 supplemented with 10% human AB serum with (PBMC_T4_BACT) or without (PBMC_T4_CTRL) 48 ug/mL sulfamethoxazole/trimethoprim (SMX-TMP). For the in vitro therapeutic experiments, PBMCs were cultured in the presence of SMX-TMP with (DRESS_Day4_TOFA) or without (DRESS_Day4_BACT) tofacitinib. The patient was treated with 10mg/d of tofacitinib, a JAK3 inhibitor. The freshly isolated PBMCs were collected again two weeks after initiation of intervention (PBMC_POST2W). Single cells from the skin, freshly isolated PBMCs, and culture PBMCs were captured using droplet based single-cell approach (10x Genomics) and library were prepared.
Results: The lymphocytes in skin and PBMCs exhibited upregulation of skin-homing chemokine receptors, CCR4 and CCR10, and JAK3 and STAT1. Treatment with tofacitinib dramatically extinguished skin inflammation in a chronic refractory case of DiHS/DRESS.
Conclusions: A successful intervention with tofacitinib in a refractory case of DiHS/DRESS was guided by the use of scRNAseq, which demonstrated aberrant activity in the JAK-STAT pathway.
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| Overall design |
Transcriptome profiles of skin, blood, and cultured PBMCs were generated by droplet-based single cell RNA-seq.
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| Contributor(s) |
Kim D, Voisin B, Nagao K |
| Citation(s) |
31959990 |
| Submission date |
Jun 16, 2019 |
| Last update date |
Feb 01, 2020 |
| Contact name |
Doyoung Kim |
| E-mail(s) |
dykim@yuhs.ac
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| Organization name |
Yonsei University College of Medicine
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| Department |
Dermatology
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| Street address |
50-1 Yonsei-ro Seodaemun-gu
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| City |
Seoul |
| ZIP/Postal code |
03722 |
| Country |
South Korea |
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| Platforms (1) |
| GPL21290 |
Illumina HiSeq 3000 (Homo sapiens) |
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| Samples (14)
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| Relations |
| BioProject |
PRJNA549083 |
| SRA |
SRP201551 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE132802_RAW.tar |
179.6 Mb |
(http)(custom) |
TAR (of H5) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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