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MYELODYSPLASTIC SYNDROME

Stress hematopoiesis induces a proliferative advantage in TET2 deficiency

Leukemia volume 36, pages 809–820 (2022)Cite this article

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Abstract

TET2 loss-of-function mutations are recurrent events in a wide range of hematological malignancies and a physiologic occurrence in blood cells of healthy older adults. It is currently unknown what determines if a person harboring a somatic TET2 mutation will progress to myelodysplastic syndrome or acute myeloid leukemia. Here we develop a zebrafish tet2 mutant through which we show that tet2 loss leads to restricted hematopoietic differentiation combined with a modest upregulation of p53, which is also characteristic of many inherited bone marrow failure syndromes. Uniquely in the context of emergency hematopoiesis by external stimuli, such as infection or cytokine stimulation, lack of tet2 leads hematopoietic stem cells to undergo excessive proliferation, resulting in an accumulation of immature cells, which are poised to become leukemogenic following additional genetic/epigenetic perturbations. This same phenomenon observed in zebrafish extends to human hematopoietic stem cells, identifying TET2 as a critical relay switch in the context of stress hematopoiesis.

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Fig. 1: The zebrafish 2.1 kb tet2 deletion mutant shows no compensation from tet1 or tet3 and is physically smaller in size.
Fig. 2: tet2 deficient zebrafish demonstrate reduced myelopoiesis.
Fig. 3: tet2 deficient zebrafish display reduced erythropoiesis.
Fig. 4:  tet2 deficient zebrafish exhibit dysplastic morphology as early as three months of age.
Fig. 5: MeDIP/hMeDIP and RNAseq analysis in adult kidney marrow show that lack of tet2 impacts hematopoietic differentiation-related genes.
Fig. 6: The absence of tet2 contributes to increased p53 protein levels but fails to induce its downstream target p21.
Fig. 7: Stress-induced granulopoiesis leads to an increase in immature cells in tet2 deficient larvae.
Fig. 8: Differentiation induction under high cytokine conditions increases BFU-E and proerythrocyte population in human HSPCs lacking TET2.

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Data and materials availability

All sequencing data have been deposited into GEO repository RNAseq (GSE136349) and meDIP/hmeDIP data (GSE136761). The codes used for meDIP-seq/hmeDIP and RNAseq analysis are uploaded to the GitHub page (https://github.com/vinothkr11/zebrafishtet2). The plasmid used for mRNA synthesis pCS2+ gcsf (Addgene ID:132947) is made available through Addgene.

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Acknowledgements

The authors would like to thank David Malloy, David Maley, Connor Booker, Gretchen Wagner, Emma Cummings for zebrafish care and maintenance, and Jennifer Curran for administrative support. We thank Dr. Todd Druley (Washington University, St. Louis) for his critical review of the paper. We would also like to extend our thanks to the Michael Smith Genome Science Centre for help with sequencing.

Funding

VR was funded by the Cancer Research Training Program of the BHCRI, with funds provided by the Terry Fox Research Institute through the Dr. Linnea Veinotte Memorial Graduate Student Award. VR was also funded by the Nova Scotia Health Research Foundation (NSHRF) Scotia Scholar award.

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Author notes

  1. Vinothkumar Rajan

    Present address: Biological Sciences Platform, Sunnybrook Research Institute, Toronto, ON, Canada

  2. These authors contributed equally: Keon Collett, Rachel Woodside.

Authors and Affiliations

  1. Department of Microbiology and Immunology, Dalhousie University, Halifax, NS, Canada

    Vinothkumar Rajan & Jason N. Berman

  2. Department of Pathology, Dalhousie University, Halifax, NS, Canada

    Keon Collett

  3. Department of Biology, Dalhousie University, Halifax, NS, Canada

    Rachel Woodside

  4. Children’s Hospital of Eastern Ontario (CHEO) Research Institute and University of Ottawa, Ottawa, ON, Canada

    Sergey V. Prykhozhij, Mira Liebman & Jason N. Berman

  5. Department of Microbiology and Immunology and Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada

    Michelle Moksa, Annäick Carles, Marcus Wong & Martin Hirst

  6. Department of Pediatrics, University of Ottawa, Ottawa, ON, Canada

    Jason N. Berman

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Contributions

VR designed and performed experiments and wrote the paper; KC, RW, and SVP designed and performed experiments and edited the paper; ML is a clinical hematopathologist and performed the hematopathology analysis. MM and AC performed MeDIP-Seq studies and edited the paper; MH oversaw the MeDIP-Seq experiment and edited the paper; JNB originated the research concept, designed experiments, and edited the paper.

Corresponding author

Correspondence to Jason N. Berman.

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JB is a member of the Scientific Advisory Board of Oxford Immune Algorithmics.

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Rajan, V., Collett, K., Woodside, R. et al. Stress hematopoiesis induces a proliferative advantage in TET2 deficiency. Leukemia 36, 809–820 (2022). https://doi.org/10.1038/s41375-021-01427-7

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