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Identification of miR-145 and miR-146a as mediators of the 5q– syndrome phenotype

Nature Medicine volume 16, pages 49–58 (2010)Cite this article

Abstract

5q– syndrome is a subtype of myelodysplastic syndrome characterized by severe anemia and variable neutropenia but normal or high platelet counts with dysplastic megakaryocytes. We examined expression of microRNAs (miRNAs) encoded on chromosome 5q as a possible cause of haploinsufficiency. We show that deletion of chromosome 5q correlates with loss of two miRNAs that are abundant in hematopoietic stem/progenitor cells (HSPCs), miR-145 and miR-146a, and we identify Toll–interleukin-1 receptor domain–containing adaptor protein (TIRAP) and tumor necrosis factor receptor–associated factor-6 (TRAF6) as respective targets of these miRNAs. TIRAP is known to lie upstream of TRAF6 in innate immune signaling. Knockdown of miR-145 and miR-146a together or enforced expression of TRAF6 in mouse HSPCs resulted in thrombocytosis, mild neutropenia and megakaryocytic dysplasia. A subset of mice transplanted with TRAF6-expressing marrow progressed either to marrow failure or acute myeloid leukemia. Thus, inappropriate activation of innate immune signals in HSPCs phenocopies several clinical features of 5q– syndrome.

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Figure 1: Expression analysis of miRNAs located on chromosome 5q in MDS.
Figure 2: Stable knockdown of miR-145 and miR-146a in mouse marrow results in a 5q–like syndrome.
Figure 3: TIRAP and TRAF6 are targets of miR-145 and miR-146a, respectively.
Figure 4: TRAF6 expression in mouse marrow results in a syndrome similar to the 5q– syndrome and progression to either marrow failure or AML.
Figure 5: Paracrine effects of IL-6 contribute to hematopoietic defects in TRAF6-transduced mice.

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Acknowledgements

We thank R. Deleeuw, R. Chari, W. Lockwood, S. Watson and E. Vucic for technical assistance and helpful discussions and M. Abbott for animal husbandry. K. Tohyama (Kawasaki Medical School) provided the MDS-L cell line. The NF-κB site–containing luciferase reporter plasmid was a gift from F. Jirik (University of Calgary). This work was supported by the Canadian Institutes of Health Research (CIHR; MOP 89976), Leukemia and Lymphoma Society of Canada and Canadian Cancer Society grants to A.K., a CIHR grant and a Terry Fox Foundation Program Project award to R.K.H., and a Stem Cell Network grant to M.M. D.T.S. is supported in part by CIHR and Michael Smith Foundation for Health Research (MSFHR) fellowships. M.M. and A.K. are Senior Scholars of the MSFHR.

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Authors and Affiliations

  1. British Columbia Cancer Agency Research Centre, Vancouver, British Columbia, Canada

    Daniel T Starczynowski, Florian Kuchenbauer, Bob Argiropoulos, Sandy Sung, Ryan Morin, Andrew Muranyi, Martin Hirst, Donna Hogge, Marco Marra, Wan Lam, R Keith Humphries & Aly Karsan

  2. Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada

    Daniel T Starczynowski, Wan Lam & Aly Karsan

  3. J.D. Crashley Myelodysplastic Syndrome Laboratory, Molecular and Cellular Biology, Sunnybrook Research Institute, Toronto, Ontario, Canada

    Richard A Wells & Rena Buckstein

  4. Department of Medicine, University of British Columbia, Vancouver, British Columbia, Canada

    R Keith Humphries

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Contributions

D.T.S. and A.K. participated in designing the research and drafting the manuscript; D.T.S. and S.S. performed experiments; F.K., B.A., A.M., and R.M. provided technical assistance and discussion; R.A.W. and R.B. provided human subject samples; D.H. and R.K.H. provided valuable advice and expertise; M.H. and M.M. provided reagents and expertise in small RNA sequencing; W.L. provided valuable reagents and expertise in array comparative genomic hybridization.

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Correspondence to Aly Karsan.

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Starczynowski, D., Kuchenbauer, F., Argiropoulos, B. et al. Identification of miR-145 and miR-146a as mediators of the 5q– syndrome phenotype. Nat Med 16, 49–58 (2010). https://doi.org/10.1038/nm.2054

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