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Rps14 haploinsufficiency causes a block in erythroid differentiation mediated by S100A8 and S100A9

Nature Medicine volume 22, pages 288–297 (2016)Cite this article

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Abstract

Impaired erythropoiesis in the deletion 5q (del(5q)) subtype of myelodysplastic syndrome (MDS) has been linked to heterozygous deletion of RPS14, which encodes the ribosomal protein small subunit 14. We generated mice with conditional inactivation of Rps14 and demonstrated an erythroid differentiation defect that is dependent on the tumor suppressor protein p53 (encoded by Trp53 in mice) and is characterized by apoptosis at the transition from polychromatic to orthochromatic erythroblasts. This defect resulted in age-dependent progressive anemia, megakaryocyte dysplasia and loss of hematopoietic stem cell (HSC) quiescence. As assessed by quantitative proteomics, mutant erythroblasts expressed higher levels of proteins involved in innate immune signaling, notably the heterodimeric S100 calcium-binding proteins S100a8 and S100a9. S100a8—whose expression was increased in mutant erythroblasts, monocytes and macrophages—is functionally involved in the erythroid defect caused by the Rps14 deletion, as addition of recombinant S100a8 was sufficient to induce a differentiation defect in wild-type erythroid cells, and genetic inactivation of S100a8 expression rescued the erythroid differentiation defect of Rps14-haploinsufficient HSCs. Our data link Rps14 haploinsufficiency in del(5q) MDS to activation of the innate immune system and induction of S100A8-S100A9 expression, leading to a p53-dependent erythroid differentiation defect.

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Figure 1: Rps14 haploinsufficiency results in a p53-mediated erythroid differentiation defect.
Figure 2: Rps14 haploinsufficiency affects HSPCs.
Figure 3: Reduced protein synthesis in Rps14-haploinsufficient cells.
Figure 4: S100a8 is upregulated in bone marrow of Rps14-haploinsufficient mice, is regulated by p53 and is necessary for the erythroid differentiation defect.
Figure 5: S100a8 is required for the erythroid-differentiation defect resulting from Rps14 haploinsufficiency.
Figure 6: S100A8+ cell frequency is increased in bone marrow from individuals with del(5q) MDS.

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Acknowledgements

This work was supported by the US National Institutes of Health (NIH) (grant no. R01HL082945; B.L.E.), a Gabrielle's Angel Award (B.L.E.), a Leukemia and Lymphoma Society Scholar and Specialized Center of Research (SCOR) award (B.L.E.), the German Research Foundation (DFG1188/3-1; R.K.S.), a Max Eder fellowship provided by the German Cancer Aid (Deutsche Krebshilfe, grant no. 111750; R.K.S.), the Edward P. Evans Foundation (R.K.S.) and the German Cluster of Excellence program Regenerative Biology to Reconstructive Therapy (REBIRTH; to G.B.). We thank D. Haase (Georg-August-Universität Göttingen) for the cytogenetic (karyotype) analysis in individuals with del(5q) MDS. This work was supported by the confocal microscope facility, a core facility of the Interdisciplinary Center for Clinical Research (Interdisziplinäres Zentrum für klinische Forschung; IZKF) Aachen, within the Faculty of Medicine at RWTH Aachen University.

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Authors and Affiliations

  1. Division of Hematology, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA

    Rebekka K Schneider, Michelle C Chen, Christopher S Waters, Edwin Chen, Lisa P Chu, R Coleman Lindsley & Benjamin L Ebert

  2. Department of Hematology, Hemostaseology, Oncology and Stem Cell Transplantation, University Hospital Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen, Aachen, Germany

    Rebekka K Schneider, Monica Ventura Ferreira, Fabian Beier, Tim H Brümmendorf & Steven A Carr

  3. Broad Institute of Harvard University and the Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Monica Schenone, Christina Hartigan & Benjamin L Ebert

  4. Renal Division, Brigham and Women's Hospital, Boston, Massachusetts, USA

    Rafael Kramann

  5. Dana-Farber Cancer Institute, Boston, Massachusetts, USA

    Cailin E Joyce, Carl D Novina & R Coleman Lindsley

  6. Department of Hematology, Oncology and Clinical Immunology, Heinrich Heine University, Düsseldorf, Germany

    Ulrich Germing

  7. Department of Medicine I, University Hospital Carl Gustav Carus, University of Technology, Dresden, Germany

    Uwe Platzbecker

  8. Institute of Pathology, Hannover Medical School, Hannover, Germany

    Guntram Büsche

  9. Institute of Pathology, University Hospital Rheinisch-Westfälische Technische Hochschule (RWTH) Aachen, Aachen, Germany

    Ruth Knüchel

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  1. Rebekka K Schneider
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Contributions

R.K.S., R.C.L., S.A.C. and B.L.E. designed experiments; R.K.S., M.S., R. Kramann, M.V.F., C.E.J., C.H., F.B., M.C.C., E.C., C.S.W., C.D.N. and L.P.C. performed experiments and analyzed data; F.B., T.H.B., U.G., U.P., R. Knüchel and G.B. collected patient samples and clinical information, reviewed bone marrow biopsies and analyzed data; R.K.S. and B.L.E. wrote the manuscript; and all authors provided critical reviews of the manuscript.

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Correspondence to Benjamin L Ebert.

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Supplementary Figures 1–7 and Supplementary Tables 1–2 (PDF 5144 kb)

Supplementary Table 3

Samples included in the proteomic analysis (XLSX 737 kb)

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Schneider, R., Schenone, M., Ferreira, M. et al. Rps14 haploinsufficiency causes a block in erythroid differentiation mediated by S100A8 and S100A9. Nat Med 22, 288–297 (2016). https://doi.org/10.1038/nm.4047

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