Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Acute myeloid leukemia

Altered expression of CSF3R splice variants impacts signal response and is associated with SRSF2 mutations

Leukemia volume 34, pages 369–379 (2020)Cite this article

Subjects

Abstract

Three annotated CSF3R mRNA splice variants have been described. CSF3R-V1 is the wild-type receptor, while CSF3R-V4 is a truncated form increased in some patients with AML. CSF3R-V3 mRNA was identified in placenta more than 20 years ago, but remains largely uncharacterized due to the lack of a suitable detection assay. Using a novel digital PCR method to quantitate expression of each CSF3R mRNA splice variant in hematopoietic cells, CSF3R-V1 was most highly expressed followed by CSF3R-V3. Functional assays revealed expression of V3 alone conferred a hypoproliferative phenotype associated with defective JAK-STAT activation. However, coexpression of V1 with V3 rescued proliferative responses. Comparative analysis of V3/V1 expression in CD34+ cells from healthy donors and patients with AML revealed a statistically significant increase in the V3/V1 ratio only in the subset of patients with AML harboring SRSF2 mutations. Knockout of SRFS2 in KG-1 and normal CD34+ cells decreased the V3/V1 ratio. Collectively, these data are the first to demonstrate expression of the CSF3R-V3 splice variant in primary human myeloid cells and a role for SRSF2 in modulating CSF3R splicing. Our findings provide confirmatory evidence that CSF3R is a target of SRSF2 mutations, which has implications for novel treatment strategies for SRSF2-mutated myeloid malignancies.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Maxson JE, Gotlib J, Pollyea DA, Fleischman AG, Agarwal A, Eide CA, et al. Oncogenic CSF3R mutations in chronic neutrophilic leukemia and atypical CML. New Engl J Med. 2013;368:1781–90.

    Article  CAS  Google Scholar 

  2. Maxson JE, Ries RE, Wang YC, Gerbing RB, Kolb EA, Thompson SL, et al. CSF3R mutations have a high degree of overlap with CEBPA mutations in pediatric AML. Blood. 2016;127:3094–8.

    Article  Google Scholar 

  3. Pardanani A, Lasho TL, Laborde RR, Elliott M, Hanson CA, Knudson RA, et al. CSF3R T618I is a highly prevalent and specific mutation in chronic neutrophilic leukemia. Leukemia. 2013;27:1870–3.

    Article  CAS  Google Scholar 

  4. Beekman R, Valkhof MG, Sanders MA, van Strien PM, Haanstra JR, Broeders L, et al. Sequential gain of mutations in severe congenital neutropenia progressing to acute myeloid leukemia. Blood. 2012;119:5071–7.

    Article  CAS  Google Scholar 

  5. Link DC, Kunter G, Kasai Y, Zhao Y, Miner T, McLellan MD, et al. Distinct patterns of mutations occurring in de novo AML versus AML arising in the setting of severe congenital neutropenia. Blood. 2007;110:1648–55.

    Article  CAS  Google Scholar 

  6. Fukunaga R, Ishizaka-Ikeda E, Pan CX, Seto Y, Nagata S. Functional domains of the granulocyte colony-stimulating factor receptor. EMBO J. 1991;10:2855–65.

    Article  CAS  Google Scholar 

  7. Avalos BR. Molecular analysis of the granulocyte colony-stimulating factor receptor. Blood. 1996;88:761–77.

    Article  CAS  Google Scholar 

  8. Dwivedi P, Greis KD. Granulocyte colony-stimulating factor receptor signaling in severe congenital neutropenia, chronic neutrophilic leukemia, and related malignancies. Exp Hematol. 2017;46:9–20.

    Article  CAS  Google Scholar 

  9. White SM, Alarcon MH, Tweardy DJ. Inhibition of granulocyte colony-stimulating factor-mediated myeloid maturation by low level expression of the differentiation-defective class IV granulocyte colony-stimulating factor receptor isoform. Blood. 2000;95:3335–40.

    Article  CAS  Google Scholar 

  10. Mehta HM, Futami M, Glaubach T, Lee DW, Andolina JR, Yang Q, et al. Alternatively spliced, truncated GCSF receptor promotes leukemogenic properties and sensitivity to JAK inhibition. Leukemia. 2014;28:1041–51.

    Article  CAS  Google Scholar 

  11. Sloand EM, Yong AS, Ramkissoon S, Solomou E, Bruno TC, Kim S, et al. Granulocyte colony-stimulating factor preferentially stimulates proliferation of monosomy 7 cells bearing the isoform IV receptor. PNAS. 2006;103:14483–8.

    Article  CAS  Google Scholar 

  12. Zhang H, Goudeva L, Immenschuh S, Schambach A, Skokowa J, Eiz-Vesper B, et al. miR-155 is associated with the leukemogenic potential of the class IV granulocyte colony-stimulating factor receptor in CD34(+) progenitor cells. Mol Med. 2015;20:736–46.

    Article  Google Scholar 

  13. Ehlers S, Herbst C, Zimmermann M, Scharn N, Germeshausen M, von Neuhoff N, et al. Granulocyte colony-stimulating factor (G-CSF) treatment of childhood acute myeloid leukemias that overexpress the differentiation-defective G-CSF receptor isoform IV is associated with a higher incidence of relapse. J Clin Oncol: Off J Am Soc Clin Oncol. 2010;28:2591–7.

    Article  CAS  Google Scholar 

  14. Fukunaga R, Ishizaka-Ikeda E, Nagata S. Growth and differentiation signals mediated by different regions in the cytoplasmic domain of granulocyte colony-stimulating factor receptor. Cell. 1993;74:1079–87.

    Article  CAS  Google Scholar 

  15. Kosmider O, Itzykson R, Chesnais V, Lasho T, Laborde R, Knudson R, et al. Mutation of the colony-stimulating factor-3 receptor gene is a rare event with poor prognosis in chronic myelomonocytic leukemia. Leukemia. 2013;27:1946–9.

    Article  CAS  Google Scholar 

  16. Tefferi A, Lasho TL, Guglielmelli P, Finke CM, Rotunno G, Elala Y, et al. Targeted deep sequencing in polycythemia vera and essential thrombocythemia. Blood Adv. 2016;1:21–30.

    Article  CAS  Google Scholar 

  17. Wong JJ, Ritchie W, Ebner OA, Selbach M, Wong JW, Huang Y, et al. Orchestrated intron retention regulates normal granulocyte differentiation. Cell. 2013;154(Aug):583–95.

    Article  CAS  Google Scholar 

  18. Cesana M, Guo MH, Cacchiarelli D, Wahlster L, Barragan J, Doulatov S, et al. A CLK3-HMGA2 alternative splicing axis impacts human hematopoietic stem cell molecular identity throughout development. Cell Stem Cell. 2018;22:575–88 e577.

    Article  CAS  Google Scholar 

  19. Nellore A, Jaffe AE, Fortin JP, Alquicira-Hernandez J, Collado-Torres L, Wang S, et al. Human splicing diversity and the extent of unannotated splice junctions across human RNA-seq samples on the sequence read archive. Genome Biol. 2016;17:266.

    Article  Google Scholar 

  20. Hermans MH, van de Geijn GJ, Antonissen C, Gits J, van Leeuwen D, Ward AC, et al. Signaling mechanisms coupled to tyrosines in the granulocyte colony-stimulating factor receptor orchestrate G-CSF-induced expansion of myeloid progenitor cells. Blood. 2003;101:2584–90.

    Article  CAS  Google Scholar 

  21. Demetri GD, Griffin JD. Granulocyte colony-stimulating factor and its receptor. Blood. 1991;78:2791–808.

    Article  CAS  Google Scholar 

  22. Berliner N, Hsing A, Graubert T, Sigurdsson F, Zain M, Bruno E, et al. Granulocyte colony-stimulating factor induction of normal human bone marrow progenitors results in neutrophil-specific gene expression. Blood. 1995;85:799–803.

    Article  CAS  Google Scholar 

  23. Hunter MG, Jacob A, O’Donnell LC, Agler A, Druhan LJ, Coggeshall KM, et al. Loss of SHIP and CIS recruitment to the granulocyte colony-stimulating factor receptor contribute to hyperproliferative responses in severe congenital neutropenia/acute myelogenous leukemia. J Immunol. 2004;173:5036–45.

    Article  CAS  Google Scholar 

  24. Hunter MG, Avalos BR. Phosphatidylinositol 3’-kinase and SH2-containing inositol phosphatase (SHIP) are recruited by distinct positive and negative growth-regulatory domains in the granulocyte colony-stimulating factor receptor. J Immunol. 1998;160:4979–87.

    CAS  PubMed  Google Scholar 

  25. Hortner M, Nielsch U, Mayr LM, Johnston JA, Heinrich PC, Haan S. Suppressor of cytokine signaling-3 is recruited to the activated granulocyte-colony stimulating factor receptor and modulates its signal transduction. J Immunol. 2002;169:1219–27.

    Article  CAS  Google Scholar 

  26. Maxson JE, Luty SB, MacManiman JD, Paik JC, Gotlib J, Greenberg P, et al. The colony-stimulating factor 3 receptor T640N mutation is oncogenic, sensitive to JAK inhibition, and mimics T618I. Clin Cancer Res. 2016;22:757–64.

    Article  CAS  Google Scholar 

  27. Smith LT, Hohaus S, Gonzalez DA, Dziennis SE, Tenen DG. PU.1 (Spi-1) and C/EBP alpha regulate the granulocyte colony-stimulating factor receptor promoter in myeloid cells. Blood. 1996;88:1234–47.

    Article  CAS  Google Scholar 

  28. Zhang DE, Hohaus S, Voso MT, Chen HM, Smith LT, Hetherington CJ, et al. Function of PU.1 (Spi-1), C/EBP, and AML1 in early myelopoiesis: regulation of multiple myeloid CSF receptor promoters. Curr Top Microbiol Immunol. 1996;211:137–47.

    CAS  PubMed  Google Scholar 

  29. Gao Y, Vasic R, Halene S. Role of alternative splicing in hematopoietic stem cells during development. Stem Cell Invest. 2018;5:26.

    Article  Google Scholar 

  30. Goldstein O, Meyer K, Greenshpan Y, Bujanover N, Feigin M, Ner-Gaon H, et al. Mapping whole-transcriptome splicing in mouse hematopoietic stem cells. Stem Cell Rep. 2017;8:163–76.

    Article  CAS  Google Scholar 

  31. Joshi P, Halene S, Abdel-Wahab O. How do messenger RNA splicing alterations drive myelodysplasia? Blood. 2017;129:2465–70.

    Article  CAS  Google Scholar 

  32. Saez B, Walter MJ, Graubert TA. Splicing factor gene mutations in hematologic malignancies. Blood. 2017;129:1260–9.

    Article  CAS  Google Scholar 

  33. Adamia S, Haibe-Kains B, Pilarski PM, Bar-Natan M, Pevzner S, Avet-Loiseau H, et al. A genome-wide aberrant RNA splicing in patients with acute myeloid leukemia identifies novel potential disease markers and therapeutic targets. Clin Cancer Res. 2014;20:1135–45.

    Article  CAS  Google Scholar 

  34. Kim E, Ilagan JO, Liang Y, Daubner GM, Lee SC, Ramakrishnan A, et al. SRSF2 mutations contribute to myelodysplasia by mutant-specific effects on exon recognition. Cancer Cell. 2015;27:617–30.

    Article  CAS  Google Scholar 

  35. Liang Y, Tebaldi T, Rejeski K, Joshi P, Stefani G, Taylor A, et al. SRSF2 mutations drive oncogenesis by activating a global program of aberrant alternative splicing in hematopoietic cells. Leukemia. 2018;32:2659–71.

    Article  CAS  Google Scholar 

  36. Menezes J, Makishima H, Gomez I, Acquadro F, Gomez-Lopez G, Grana O, et al. CSF3R T618I co-occurs with mutations of splicing and epigenetic genes and with a new PIM3 truncated fusion gene in chronic neutrophilic leukemia. Blood Cancer J. 2013;3:e158.

    Article  CAS  Google Scholar 

  37. Zhang J, Lieu YK, Ali AM, Penson A, Reggio KS, Rabadan R, et al. Disease-associated mutation in SRSF2 misregulates splicing by altering RNA-binding affinities. PNAS. 2015;112:E4726–4734.

    Article  CAS  Google Scholar 

  38. Pellagatti A, Armstrong RN, Steeples V, Sharma E, Repapi E, Singh S, et al. Impact of spliceosome mutations on RNA splicing in myelodysplasia: dysregulated genes/pathways and clinical associations. Blood. 2018;132:1225–40.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported in part by The Kerry and Simone Vickar Family Foundation and the Leon Levine Foundation.

Author information

Authors and Affiliations

  1. The Department of Hematologic Oncology and Blood Disorders, The Levine Cancer Institute; Atrium Health, Charlotte, NC, USA

    Amanda Lance, Lawrence J. Druhan, C. Greer Vestal, Andrea Price, Elise Tjaden, Andee N. Fox & Belinda R. Avalos

  2. The Molecular Biology and Genomics Laboratory, The Levine Cancer Institute; Atrium Health, Charlotte, NC, USA

    Nury M. Steuerwald, Alicia Hamilton & Mathew Smith

Authors
  1. Amanda Lance
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lawrence J. Druhan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Greer Vestal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nury M. Steuerwald
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alicia Hamilton
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mathew Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Andrea Price
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Elise Tjaden
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Andee N. Fox
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Belinda R. Avalos
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

AL and CGV designed and performed experiments, analyzed data, and contributed to writing the manuscript. LJD and BRA designed experiments, analyzed data, and wrote the manuscript. AH, MS, AP, and NMS designed and performed experiments and analyzed data. ET and ANF collected data. All authors assisted in editing the manuscript.

Corresponding author

Correspondence to Belinda R. Avalos.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lance, A., Druhan, L.J., Vestal, C.G. et al. Altered expression of CSF3R splice variants impacts signal response and is associated with SRSF2 mutations. Leukemia 34, 369–379 (2020). https://doi.org/10.1038/s41375-019-0567-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41375-019-0567-9

This article is cited by

Search

Advanced search

Quick links