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| Status |
Public on Jan 03, 2020 |
| Title |
Prospective isolation of radiation induced erythroid stress progenitors reveals unique transcriptomic and epigenetic signatures enabling increased erythroid output |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
Massive expansion of erythroid progenitor cells is essential for surviving anemic stress. Research towards understanding this critical process, referred to as stress-erythropoiesis, has been hampered due to lack of specific marker-combinations enabling analysis of the distinct stress-progenitor cells capable of providing radioprotection and enhanced red blood cell production. Here we present a method for precise identification and in vivo validation of progenitor cells contributing to both steady-state and stress-erythropoiesis, enabling for the first time in-depth molecular characterization of these cells. Differential expression of surface markers CD150, CD9 and Sca1 defines a hierarchy of splenic stress-progenitors during irradiation-induced stress recovery in mice, and provides high-purity isolation of the functional stress-BFU-Es with a 100-fold improved enrichment compared to state-of-the-art. By transplanting purified stress-progenitors expressing the fluorescent protein Kusabira Orange, we have for the first time determined their kinetics in vivo and demonstrated that CD150+CD9+Sca1- stress-BFU-Es provide a massive but transient radioprotective erythroid wave, followed by multi-lineage reconstitution from CD150+CD9+Sca1+ multi-potent stem/progenitor cells. Whole genome transcriptional analysis revealed that stress-BFU-Es express gene signatures more associated with erythropoiesis and proliferation compared to steady-state BFU-Es, and are BMP-responsive. Evaluation of chromatin accessibility through ATAC sequencing reveals enhanced and differential accessibility to binding sites of the chromatin-looping transcription factor CTCF in stress-BFU-Es compared to steady-state BFU-Es. Our findings offer molecular insight to the unique capacity of stress-BFU-Es to rapidly form erythroid cells in response to anemia and constitute an important step towards identifying novel erythropoiesis stimulating agents.
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| Overall design |
For RNA-seq, one steady-state BFU-E population in bone marrow and one from irradiation-induced stress erythropoiesis in spleen were sorted and sequenced in triplicates together with one stressed splenic MPP and one CFU-E population for internal comparison. For ATAC-seq, one steady-state BFU-E population in bone marrow and one from irradiation-induced stress erythropoiesis in spleen were sorted and sequenced in triplicates.
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| Contributor(s) |
Singbrant S, Mattebo A, Strid T, Sigvardsson M, Flygare J |
| Citation(s) |
33131245 |
| |
| Submission date |
Oct 24, 2019 |
| Last update date |
Mar 01, 2021 |
| Contact name |
Johan Flygare |
| E-mail(s) |
johan.flygare@med.lu.se
|
| Organization name |
Lund University
|
| Lab |
Flygare
|
| Street address |
Sölvegatan 19
|
| City |
Lund |
| ZIP/Postal code |
22184 |
| Country |
Sweden |
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| Platforms (2) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
| GPL21493 |
Illumina HiSeq 3000 (Mus musculus) |
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| Samples (18)
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| Relations |
| BioProject |
PRJNA579342 |
| SRA |
SRP226834 |
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