Abstract
Alternatively spliced colony stimulating factor 3 receptor (CSF3R) isoforms Class III and Class IV are observed in myelodysplastic syndromes (MDS), but their roles in disease remain unclear. We report that the MDS-associated splicing factor SRSF2 affects the expression of Class III and Class IV isoforms and perturbs granulopoiesis. Add-back of the Class IV isoform in Csf3r-null mouse progenitor cells increased granulocyte progenitors with impaired neutrophil differentiation, while add-back of the Class III produced dysmorphic neutrophils in fewer numbers. These CSF3R isoforms were elevated in patients with myeloid neoplasms harboring SRSF2 mutations. Using in vitro splicing assays, we confirmed increased Class III and Class IV transcripts when SRSF2 P95 mutations were co-expressed with the CSF3R minigene in K562 cells. Since SRSF2 regulates splicing partly by recognizing exonic splicing enhancer (ESE) sequences on pre-mRNA, deletion of either ESE motifs within CSF3R exon 17 decreased Class IV transcript levels without affecting Class III. CD34+ cells expressing SRSF2 P95H showed impaired neutrophil differentiation in response to G-CSF and was accompanied by increased levels of Class IV. Our findings suggest that SRSF2 P95H promotes Class IV splicing by binding to key ESE sequences in CSF3R exon 17, and that SRSF2, when mutated, contributes to dysgranulopoiesis.
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Acknowledgements
This work was supported by DOD Bone Marrow Failure Grant W81XWH-15-1-0153 (SJC), NIH R01-HL128173 (SJC, MK), and VeloSano Award (HMM). MK was supported by the National Science Centre (Poland) grant 2018/29/B/ST7/02550. Department of Systems Biology and Engineering, Silesian University of Technology, Poland. Juan Sabín, co-founder of Affinimeter, assisted the authors in curve fitting the ITC data. The data set used from Hershberger et al. [32] was supported by funds from “Torsten Haferlach Leukämiediagnostik Stiftung” (https://www.torsten-haferlach-leukaemiediagnostik-stiftung.de/en/).
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Design of experiments (BAW, HMM, SRP, CC, BST, SJC), the performance of experiments (BAW, HMM, SRP, TH), analysis of data (BAW, HMM, SRP, BST, MK, JPM, SJC), and writing of manuscript (BAW, SJC).
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TH serves on the management board of the Munich Leukemia Laboratory. All of the other authors declare no conflict of interest.
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Wang, B.A., Mehta, H.M., Penumutchu, S.R. et al. Alternatively spliced CSF3R isoforms in SRSF2 P95H mutated myeloid neoplasms. Leukemia 36, 2499–2508 (2022). https://doi.org/10.1038/s41375-022-01672-4
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DOI: https://doi.org/10.1038/s41375-022-01672-4
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