Runx factors launch T cell and innate lymphoid programs via direct and gene network-based mechanisms
Abstract
Runx factors are essential for lineage specification of various hematopoietic cells, including T lymphocytes. However, they regulate context-specific genes and occupy distinct genomic regions in different cell types. Here, we show that dynamic Runx binding shifts in mouse early T cell development are mostly not restricted by local chromatin state but regulated by Runx dosage and functional partners. Runx cofactors compete to recruit a limited pool of Runx factors in early T progenitor cells, and a modest increase in Runx protein availability at pre-commitment stages causes premature Runx occupancy at post-commitment binding sites. This increased Runx factor availability results in striking T cell lineage developmental acceleration by selectively activating T cell-identity and innate lymphoid cell programs. These programs are collectively regulated by Runx together with other, Runx-induced transcription factors that co-occupy Runx-target genes and propagate gene network changes.
Copyright and License
© The Author(s), under exclusive licence to Springer Nature America, Inc. 2023, corrected publication 2023.
Acknowledgement
We thank the E.V.R. laboratory members for helpful discussions, R. Diamond and members of the Caltech Flow Cytometry and Cell Sorting facility for sorting, I. Antoshechkin and V. Kumar of the Caltech Jacobs Genomics Facility for sequencing, H. Amrhein and D. Trout for computer support, J. Park and S. Chen from the Caltech Single Cell Profiling and Engineering Center for support for processing 10x Chromium samples, J. Vielmetter and the Caltech Protein Expression Center for purifying protein A-MNase, I. Soto for mouse care, M. Quiloan and M. Chau for mouse genotyping and supervision, and J. Longmate (formerly City of Hope) and M. Yui (formerly Caltech) for statistics advice. Support for this project came from USPHS grants (R01AI135200, R01HL119102 and R01HD100039) to E.V.R., and from a Cancer Research Institute Irvington Postdoctoral Fellowship CRI.SHIN and Caltech Baxter Fellowship (to B.S.). F.G. was supported in part by the National Institutes of Health award 1RF1NS122060-01. Support also came from The Beckman Institute at Caltech for all the Caltech facilities and from the Biology and Biological Engineering Division Bowes Leadership Chair Fund, the Louis A. Garfinkle Memorial Laboratory Fund and the Al Sherman Foundation to the E.V.R. laboratory. E.V.R. gratefully acknowledges support from the Edward B. Lewis Professorship and past support from the Albert Billings Ruddock Professorship.
Contributions
B.S. and E.V.R. conceptualized the project, wrote the paper and edited the paper. B.S. performed the experiments, and analyzed data with W.Z. and J.W. F.G. wrote the in-house bioinformatic pipeline for hashtag alignment and provided further analysis. E.V.R. supervised research, acquired funding and provided additional data analysis. All authors edited the paper and provided helpful comments.
Data Availability
All new genomic sequencing data have been deposited in Gene Expression Omnibus under accession numbers GSE218147 (C&R, ChIP–seq and ATAC-seq) and GSE218149 (scRNA-seq). The publicly available data utilized for analysis are presented in Supplementary Table 7. Publicly available data were utilized by downloading raw sequence read files using the Sequence Read Archive toolkit Fastq-dump (v.2.10.9). Source data are provided with this paper. All other data needed to evaluate the conclusions in the paper are present in the paper, the Extended Data or the Supplementary Information, or are available upon request.
Code Availability
All code used for data analysis in this work is publicly available and listed in the Methods and Reporting Summary.
Additional Information
Change history
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27 November 2023
A Correction to this paper has been published: https://doi.org/10.1038/s41590-023-01716-6
Conflict of Interest
W.Z. is employed by BillionToOne and has been employed by 10x Genomics (CA 94588). F.G. is employed by Lyterian Therapeutics. E.V.R. was a member of the Scientific Advisory Board for Century Therapeutics and has advised Kite Pharma and A2 Biotherapeutics. The remaining authors declare no competing interests.
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Additional details
- PMCID
- PMC10673614
- DOI
- 10.1038/s41590-023-01585-z
- National Institutes of Health
- R01AI135200
- National Institutes of Health
- R01HL119102
- National Institutes of Health
- R01HD100039
- Cancer Research Institute
- Irvington Postdoctoral Fellowship CRI.SHIN
- National Institutes of Health
- 1RF1NS122060-01
- California Institute of Technology
- Beckman Institute
- California Institute of Technology
- Biology and Biological Engineering Division Bowes Leadership Chair Fund
- California Institute of Technology
- Louis A. Garfinkle Memorial Laboratory Fund
- California Institute of Technology
- Edward B. Lewis Professorship
- California Institute of Technology
- Albert Billings Ruddock Professorship
- Caltech groups
- Division of Biology and Biological Engineering, Millard and Muriel Jacobs Genetics and Genomics Laboratory