Journal
ALLERGY
Volume 76, Issue 6, Pages 1731-1742Publisher
WILEY
DOI: 10.1111/all.14633
Keywords
basophils; differentiation; mast cells; single‐ cell RNA sequencing; transcriptomics
Categories
Funding
- Swedish Research Council [2015-06322, 2018-02070]
- Swedish Cancer Society
- Magnus Bergvall Foundation
- Lars Hierta Memorial Foundation
- Karolinska Institutet
- Blood Cancer UK [18002]
- Wellcome [206328/Z/17/Z, 100140/Z/12/Z]
- CRUK [C1163/A21762]
- NIH-NIDDK [1 R24 DK106766]
- MRC [MR/S036113/1, 203151/Z/16/Z]
- MRC Physical Biology of Stem Cells PhD studentship [MR/K500975/1]
- Wellcome Strategic Award [105031/D/14/Z]
- Angstromke Wiberg Foundation
- Swedish Research Council [2015-06322, 2018-02070] Funding Source: Swedish Research Council
- Wellcome Trust [105031/D/14/Z, 206328/Z/17/Z, 100140/Z/12/Z] Funding Source: Wellcome Trust
- MRC [MR/S036113/1] Funding Source: UKRI
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The study explored the differentiation pathways of basophils and mast cells at the single-cell level in mice using multicolor flow cytometry, high-coverage single-cell RNA sequencing, and cell fate assays. The results provided a detailed map of differentiation and revealed gene expression changes during the process. Flow cytometry and transcriptional profiling successfully predicted the bipotent phenotype of a previously uncharacterized population of peritoneal basophil-mast cell progenitors.
Background Basophils and mast cells contribute to the development of allergic reactions. Whereas these mature effector cells are extensively studied, the differentiation trajectories from hematopoietic progenitors to basophils and mast cells are largely uncharted at the single-cell level. Methods We performed multicolor flow cytometry, high-coverage single-cell RNA sequencing analyses, and cell fate assays to chart basophil and mast cell differentiation at single-cell resolution in mouse. Results Analysis of flow cytometry data reconstructed a detailed map of basophil and mast cell differentiation, including a bifurcation of progenitors into two specific trajectories. Molecular profiling and pseudotime ordering of the single cells revealed gene expression changes during differentiation. Cell fate assays showed that multicolor flow cytometry and transcriptional profiling successfully predict the bipotent phenotype of a previously uncharacterized population of peritoneal basophil-mast cell progenitors. Conclusions A combination of molecular and functional profiling of bone marrow and peritoneal cells provided a detailed road map of basophil and mast cell development. An interactive web resource was created to enable the wider research community to explore the expression dynamics for any gene of interest.
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