Abstract
The miR17~92 cluster plays important roles in haematopoiesis. However, it is not clear at what stage of differentiation and through which targets miR17~92 exerts this function. Therefore, we generated miR17~92fl/fl; RosaCreERT2 mice for inducible deletion of miR17~92 in haematopoietic cells. Bone marrow reconstitution experiments revealed that miR17~92-deleted cells were not capable to contribute to mature haematopoietic lineages, which was due to defects in haematopoietic stem/progenitor cells (HSPCs). To identify the critical factor targeted by miR17~92 we performed gene expression analysis in HSPCs, demonstrating that mRNA levels of pro-apoptotic Bim inversely correlated with the expression of the miR17~92 cluster. Strikingly, loss of pro-apoptotic BIM completely prevented the loss of HSPCs caused by deletion of miR17~92. The BIM/miR17~92 interaction is conserved in human CD34+ HSPCs, as miR17~92 inhibition or blockade of its binding to the BIM 3′UTR reduced the survival and growth of these cells. Despite the prediction that miR17~92 functions by impacting a plethora of different targets, the absence of BIM alone is sufficient to prevent all defects caused by deletion of miR17~92 in haematopoietic cells.
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Acknowledgements
We thank C Stivala, S Russo, J Mansheim, T Baldinger, M Patsis and G Siciliano for expert animal care; B Helbert and K Mackwell for genotyping; J Corbin and J McManus for automated blood analysis; S Monard and his team for help with flow cytometry; S Mifsud for assistance with colony assays and Dr P Bouillet for providing Bim−/− mice. This work was supported by grants and fellowships from the Deutsche Krebshilfe (Dr Mildred Scheel post-doctoral fellowship to KB) the Australian National Health and Medical Research Council (NHMRC) (Project Grants 1145728 to MJH, 1143105 to MJH and AS, 1122783 to APN, Program Grants 1016701 to AS and 1113577 to WSA and Fellowships 1020363 to AS, 1058344 to WSA, 1156095 to MJH, GNT1035229 to CAdG and 1124788 to NDH), the Leukemia and Lymphoma Society of America (LLS SCOR 7001-13 to AS and MJH), the Cancer Council of Victoria (project grant 1052309 to AS and Venture Grant to MJH and AS), a Victorian Cancer Agency Fellowship (ECSG18020 to JR) as well as by operational infrastructure grants through the Australian Government Independent Research Institute Infrastructure Support Scheme (361646 and 9000220) and the Victorian State Government Operational Infrastructure Support Program.
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KB, MJH and AS designed and conceived the study, planned experiments and prepared the paper. KB conducted and analysed the experiments. CAdG helped with the analysis of the RNA expression data (Fig. 1b) CH and WSA provided advice for the analysis of the LT-HSC compartment, helped with these experiments and data analysis and provided the antibodies for staining cells (Figs. 3–5). APN and LDR performed the colony formation assays (Fig. 6a). JR and NDH gave advice on experiments with human CD34+ cord blood cells and provided the cells and media (Fig. 6). EM provided MIR17PTi and helped with planning the colony formation assays (Fig. 6a).
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Brinkmann, K., Ng, A.P., de Graaf, C.A. et al. miR17~92 restrains pro-apoptotic BIM to ensure survival of haematopoietic stem and progenitor cells. Cell Death Differ 27, 1475–1488 (2020). https://doi.org/10.1038/s41418-019-0430-6
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DOI: https://doi.org/10.1038/s41418-019-0430-6
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