The n6-methyladenosine (m6a)-forming enzyme mettl3 controls myeloid differentiation of normal hematopoietic and leukemia cells

ABSTRACT _N_6-methyladenosine (m6A) is an abundant nucleotide modification in mRNA that is required for the differentiation of mouse embryonic stem cells. However, it remains unknown whether

the m6A modification controls the differentiation of normal and/or malignant myeloid hematopoietic cells. Here we show that shRNA-mediated depletion of the m6A-forming enzyme METTL3 in

human hematopoietic stem/progenitor cells (HSPCs) promotes cell differentiation, coupled with reduced cell proliferation. Conversely, overexpression of wild-type METTL3, but not of a

catalytically inactive form of METTL3, inhibits cell differentiation and increases cell growth. _METTL3_ mRNA and protein are expressed more abundantly in acute myeloid leukemia (AML) cells

than in healthy HSPCs or other types of tumor cells. Furthermore, METTL3 depletion in human myeloid leukemia cell lines induces cell differentiation and apoptosis and delays leukemia

progression in recipient mice _in vivo_. Single-nucleotide-resolution mapping of m6A coupled with ribosome profiling reveals that m6A promotes the translation of _c-MYC_, _BCL2_ and _PTEN_

mRNAs in the human acute myeloid leukemia MOLM-13 cell line. Moreover, loss of METTL3 leads to increased levels of phosphorylated AKT, which contributes to the differentiation-promoting

effects of METTL3 depletion. Overall, these results provide a rationale for the therapeutic targeting of METTL3 in myeloid leukemia. Access through your institution Buy or subscribe This is

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PROGRESSION BY ENHANCING RCC2 STABILITY Article Open access 25 February 2022 NEW INSIGHT INTO THE CATALYTIC -DEPENDENT AND -INDEPENDENT ROLES OF METTL3 IN SUSTAINING ABERRANT TRANSLATION IN

CHRONIC MYELOID LEUKEMIA Article Open access 24 September 2021 YTHDC1-MEDIATED MICRORNA MATURATION IS ESSENTIAL FOR HEMATOPOIETIC STEM CELLS MAINTENANCE Article Open access 16 October 2024

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RNA-seq data with DESeq2. _Genome Biol._ 15, 550 (2014). Article  Google Scholar  Download references ACKNOWLEDGEMENTS We thank D. Bachovchin (MSKCC, New York) for the MOLM-13 constitutively

expressing Cas9 cell line, and C. Vakoc (Cold Spring Harbor Laboratory, New York) for the constitutively expressing Cas9-RN2c cell line. We would like to thank the members of the Scaltriti

laboratory (MSKCC, New York) for providing us with PI3K/AKT inhibitors. We thank the Weill Cornell Medicine Epigenomics Core for their assistance with sequencing. M.G.K. was supported by the

US National Institutes of Health National Institute of Diabetes Digestive and Kidney Diseases Career Development Award, NIDDK NIH R01-DK101989-01A1, NCI 1R01CA193842-01, Kimmel Scholar

Award, V-Scholar Award, Geoffrey Beene Award, Leukemia Lymphoma Society Career Development Award and Alex's Lemonade Stand A Award. This work was also supported by a Tri-Institutional

Stem Cell Award (M.G.K. and S.R.J.), R01CA186702 (S.R.J.), T32CA062948 (B.F.P.), Ruth L. Kirschstein National Research Service Award 1F32CA22104-01 (B.F.P.), a Damon Runyon-Sohn Pediatric

Cancer Fellowship Award DRSG10-14 (L.P.V.), and the American-Italian Cancer Foundation (S.Z.). The research was funded in part through the NIH/NCI Cancer Support Core Grant P30 CA08748 MGK.

The RPPA core facility is funded by NCI #CA16672. AUTHOR INFORMATION Author notes * Ly P Vu, Brian F Pickering and Yuanming Cheng: These authors contributed equally to this work. AUTHORS AND

AFFILIATIONS * Molecular Pharmacology Program, Center for Cell Engineering, Center for Stem Cell Biology, Center for Experimental Therapeutics, Center for Hematologic Malignancies, Memorial

Sloan Kettering Cancer Center, New York, New York, USA Ly P Vu, Yuanming Cheng, Diu Nguyen, Gerard Minuesa, Timothy Chou, Arthur Chow & Michael G Kharas * Department of Pharmacology,

Weill Cornell Medicine, Cornell University, New York, New York, USA Brian F Pickering, Sara Zaccara & Samie R Jaffrey * Department of Physiology and Biophysics, Weill Cornell Medicine,

Cornell University, New York, New York, USA Yogesh Saletore, Matthew MacKay & Christopher E Mason * Department of Medicine, Hematologic Oncology Tissue Bank, Memorial Sloan Kettering

Cancer Center, New York, New York, USA Jessica Schulman * Center for Hematologic Malignancies, Memorial Sloan Kettering Cancer Center, New York, New York, USA Christopher Famulare & 

Minal Patel * Department of Medicine, Memorial Sloan Kettering Cancer Center, Leukemia Service, New York, New York, USA Virginia M Klimek * Department of Medicine and Department of

Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, Virginia, USA Francine E Garrett-Bakelman * Division of Hematology and Medical Oncology, Department of Medicine

and Department of Pharmacology, Weill Cornell Medicine, Cornell University, New York, New York, USA Ari Melnick * Division of Hematology and Oncology, University of Pennsylvania,

Philadelphia, Pennsylvania, USA Martin Carroll * The HRH Prince Alwaleed Bin Talal Bin Abdulaziz Alsaud Institute for Computational Biomedicine, Weill Cornell Medicine, New York, New York,

USA Christopher E Mason * The Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, New York, New York, USA Christopher E Mason Authors * Ly P Vu View author publications

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project, performed experiments, analyzed data and wrote the manuscript. B.F.P., Y.C., D.N. and S.Z. performed experiments, analyzed data and edited the manuscript. G.M., T.C. and A.C.

provided experimental supports. C.E.M., Y.S. and M.M. performed and analyzed MeRIP-seq experiments on patient-derived samples. J.S., C.F., M.P., F.E.G.-B., A.M., V.M.K. and M.C. provided

patient samples. S.R.J. supervised the project and wrote the manuscript. M.G.K. directed the project, analyzed data and wrote the manuscript. Publisher's note: Springer Nature remains

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DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY FIGURES Supplementary Figures 1–6 (PDF 1925 kb) LIFE SCIENCES

REPORTING SUMMARY (PDF 86 KB) SUPPLEMENTARY DATASET Supplementary Tables 1–13 (XLSX 9967 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Vu, L.,

Pickering, B., Cheng, Y. _et al._ The _N_6-methyladenosine (m6A)-forming enzyme METTL3 controls myeloid differentiation of normal hematopoietic and leukemia cells. _Nat Med_ 23, 1369–1376

(2017). https://doi.org/10.1038/nm.4416 Download citation * Received: 07 May 2017 * Accepted: 06 September 2017 * Published: 18 September 2017 * Issue Date: 01 November 2017 * DOI:

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