METTL16 drives leukemogenesis and leukemia stem cell self-renewal by reprogramming BCAA metabolism.
N6-methyladenosine (m6A), the most prevalent internal modification in mammalian mRNAs, is involved in many pathological processes. METTL16 is a recently identified m6A methyltransferase. However, its role in leukemia has yet to be investigated. Here, we show that METTL16 is a highly essential gene for the survival of acute myeloid leukemia (AML) cells via CRISPR-Cas9 screening and experimental validation. METTL16 is aberrantly overexpressed in human AML cells, especially in leukemia stem cells (LSCs) and leukemia-initiating cells (LICs). Genetic depletion of METTL16 dramatically suppresses AML initiation/development and maintenance and significantly attenuates LSC/LIC self-renewal, while moderately influencing normal hematopoiesis in mice. Mechanistically, METTL16 exerts its oncogenic role by promoting expression of branched-chain amino acid (BCAA) transaminase 1 (BCAT1) and BCAT2 in an m6A-dependent manner and reprogramming BCAA metabolism in AML. Collectively, our results characterize the METTL16/m6A/BCAT1-2/BCAA axis in leukemogenesis and highlight the essential role of METTL16-mediated m6A epitranscriptome and BCAA metabolism reprograming in leukemogenesis and LSC/LIC maintenance.
Pubmed ID: 36608679 RIS Download
Research resources used in this publication
- HEK293T (RRID:CVCL_0063) (cell line)
- Kasumi-1 (RRID:CVCL_0589) (cell line)
- NOMO-1 (RRID:CVCL_1609) (cell line)
- NB4 (RRID:CVCL_0005) (cell line)
-
NOD.Cg-Rag1tm1Mom Il2rg
- pCL-Eco (RRID:Addgene_12371) (plasmid)
- lentiCas9-Blast (RRID:Addgene_52962) (plasmid)
- lenti-sgRNA hygro (RRID:Addgene_104991) (plasmid)
- pLKO.1 puro (RRID:Addgene_8453) (plasmid)
- psPAX2 (RRID:Addgene_12260) (plasmid)
- pMD2.G (RRID:Addgene_12259) (plasmid)
- pLenti CMV Puro LUC (w168-1) (RRID:Addgene_17477) (plasmid)
- B6.Cg-Tg(Mx1-cre)1Cgn/J (RRID:IMSR_JAX:003556) (organism)
-
B6.SJL-PtprcaPepc
- OP9 (RRID:CVCL_4398) (cell line)
- Mono-Mac-6 (RRID:CVCL_1426) (cell line)
- ML-2 (RRID:CVCL_1418) (cell line)
- MOLM-13 (RRID:CVCL_2119) (cell line)
- HL-60(TB) (RRID:CVCL_A794) (cell line)
- THP-1 (RRID:CVCL_0006) (cell line)
-
NOD.Cg-Rag1tm1Mom Il2rg
Additional research tools detected in this publication
None foundAntibodies used in this publication
- TER-119/Erythroid cells (RRID:AB_10563222)
- CD11b-Biotin, human and mouse (RRID:AB_2660137)
- Anti-Gr-1-Biotin (RRID:AB_2659872)
- CD45.1 Monoclonal Antibody (A20), PE, eBioscience (RRID:AB_465675)
- FITC anti-mouse CD45.2 (RRID:AB_313443)
- CD45.2 Monoclonal Antibody (104), APC, eBioscience (RRID:AB_469400)
- Rat Anti-CD11b Monoclonal Antibody, Allophycocyanin Conjugated, Clone M1/70 (RRID:AB_398535)
- Ly-6G/Ly-6C Monoclonal Antibody (RB6-8C5), PE, eBioscience (RRID:AB_466044)
- CD11b Monoclonal Antibody (M1/70), PerCP-Cyanine5.5, eBioscience (RRID:AB_953558)
- CD45R/B220 (RRID:AB_10561687)
- APC anti-mouse CD3 (RRID:AB_2561456)
- CD48 Monoclonal Antibody (HM48-1), FITC, eBioscience (RRID:AB_465077)
- CD115 (c-fms) Monoclonal Antibody (AFS98), PerCP-eFluor™ 710, eBioscience (RRID:AB_10597740)
- PE/Cyanine7 anti-mouse CD34 (RRID:AB_2716078)
- APC/Cyanine7 anti-mouse CD16/32 (RRID:AB_2800710)
- Ly-6A/E (Sca-1) Monoclonal Antibody (D7), PE, eBioscience (RRID:AB_466086)
- CD117 (c-Kit) Monoclonal Antibody (2B8), APC, eBioscience (RRID:AB_469431)
- Mouse Hematopoietic Lineage Antibody Cocktail, eFluor™ 450, eBioscience (RRID:AB_10426799)
- CD14 Monoclonal Antibody (61D3), APC, eBioscience (RRID:AB_10669167)
- CD11b Monoclonal Antibody (ICRF44), PE, eBioscience (RRID:AB_2043799)
- Anti-rabbit IgG (H+L), F(ab) 2 Fragment (Alexa Fluor 555 Conjugate) (RRID:AB_10694110)
- Anti-Puromycin Antibody, clone 12D10 (RRID:AB_2566826)
- CD34 Monoclonal Antibody (4H11), FITC, eBioscience (RRID:AB_1518732)
- BV786 Mouse Anti-Human CD45 (RRID:AB_2716864)
- Goat Anti-Mouse IgG - H&L Polyclonal Antibody, HRP Conjugated (RRID:AB_955439)
- Goat Anti-Rabbit IgG - H&L Polyclonal antibody, Hrp Conjugated (RRID:AB_955447)
- Normal Mouse IgG (RRID:AB_145840)
- Monoclonal ANTI-FLAG® M2 antibody (RRID:AB_259529)
- vinculin (H-10) (RRID:AB_628438)
- β-Actin (8H10D10) Mouse mAb (RRID:AB_2242334)
- LARS antibody (RRID:AB_10733878)
- BCAT2 antibody (RRID:AB_10792411)
- BCAT1 mouse monoclonal antibody,clone 3F5 (RRID:AB_11126287)
- Anti-METT10D antibody produced in rabbit (RRID:AB_1853828)
- Rabbit anti-METTL16 Antibody, Affinity Purified (RRID:AB_2620389)
- Anti-m6A (RRID:AB_2279214)
Associated grants
- Agency: NCI NIH HHS, United States
Id: R01 CA236399 - Agency: NCI NIH HHS, United States
Id: R50 CA211404 - Agency: NIDDK NIH HHS, United States
Id: R01 DK124116 - Agency: NCI NIH HHS, United States
Id: R01 CA214965 - Agency: NCI NIH HHS, United States
Id: R01 CA211614 - Agency: NCI NIH HHS, United States
Id: R37 CA233691 - Agency: NCI NIH HHS, United States
Id: R01 CA243386 - Agency: NCI NIH HHS, United States
Id: R01 CA236626 - Agency: NCI NIH HHS, United States
Id: T32 CA186895 - Agency: NCI NIH HHS, United States
Id: R01 CA271497
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TER-119/Erythroid cells (antibody)
RRID:AB_10563222
This monoclonal targets TER-119/Erythroid cells
View all literature mentionsCD11b-Biotin, human and mouse (antibody)
RRID:AB_2660137
This monoclonal targets CD11b
View all literature mentionsAnti-Gr-1-Biotin (antibody)
RRID:AB_2659872
This monoclonal targets Gr-1
View all literature mentionsCD45.1 Monoclonal Antibody (A20), PE, eBioscience (antibody)
RRID:AB_465675
This monoclonal targets CD45.1
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RRID:AB_313443
This monoclonal targets CD45.2
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RRID:AB_469400
This monoclonal targets CD45.2
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RRID:AB_398535
This monoclonal targets CD11b
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RRID:AB_466044
This monoclonal targets Ly-6G/Ly-6C
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RRID:AB_953558
This monoclonal targets CD11b
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RRID:AB_10561687
This monoclonal targets CD45R (B220)
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RRID:AB_2561456
This monoclonal targets CD3
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RRID:AB_465077
This monoclonal targets CD48
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RRID:AB_10597740
This monoclonal targets CD115 (c-fms)
View all literature mentionsPE/Cyanine7 anti-mouse CD34 (antibody)
RRID:AB_2716078
This monoclonal targets CD34
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This monoclonal targets CD16/32
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RRID:AB_466086
This monoclonal targets Ly-6A/E (Sca-1)
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RRID:AB_469431
This monoclonal targets CD117 (c-Kit)
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RRID:AB_10426799
This cocktail targets Hematopoietic Lineage
View all literature mentionsCD14 Monoclonal Antibody (61D3), APC, eBioscience (antibody)
RRID:AB_10669167
This monoclonal targets CD14
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RRID:AB_2043799
This monoclonal targets CD11b
View all literature mentionsAnti-rabbit IgG (H+L), F(ab) 2 Fragment (Alexa Fluor 555 Conjugate) (antibody)
RRID:AB_10694110
This unknown targets rabbit IgG (H+L) F(ab) 2 Fragment
View all literature mentionsAnti-Puromycin Antibody, clone 12D10 (antibody)
RRID:AB_2566826
This monoclonal targets Puromycin from Streptomyces alboniger
View all literature mentionsCD34 Monoclonal Antibody (4H11), FITC, eBioscience (antibody)
RRID:AB_1518732
This monoclonal targets CD34
View all literature mentionsBV786 Mouse Anti-Human CD45 (antibody)
RRID:AB_2716864
This monoclonal targets CD45
View all literature mentionsGoat Anti-Mouse IgG - H&L Polyclonal Antibody, HRP Conjugated (antibody)
RRID:AB_955439
This polyclonal targets Mouse Mouse IgG secondary - H&L
View all literature mentionsGoat Anti-Rabbit IgG - H&L Polyclonal antibody, Hrp Conjugated (antibody)
RRID:AB_955447
This polyclonal targets Rabbit Rabbit IgG secondary - H&L
View all literature mentionsNormal Mouse IgG (antibody)
RRID:AB_145840
This polyclonal secondary targets not applicable
View all literature mentionsMonoclonal ANTI-FLAG® M2 antibody (antibody)
RRID:AB_259529
This monoclonal targets FLAG
View all literature mentionsvinculin (H-10) (antibody)
RRID:AB_628438
This monoclonal targets Human VCL
View all literature mentionsβ-Actin (8H10D10) Mouse mAb (antibody)
RRID:AB_2242334
This monoclonal targets beta-actin
View all literature mentionsBCAT2 antibody (antibody)
RRID:AB_10792411
This polyclonal targets BCAT2
View all literature mentionsBCAT1 mouse monoclonal antibody,clone 3F5 (antibody)
RRID:AB_11126287
This monoclonal targets BCAT1
View all literature mentionsAnti-METT10D antibody produced in rabbit (antibody)
RRID:AB_1853828
This unknown targets Human METT10D
View all literature mentionsRabbit anti-METTL16 Antibody, Affinity Purified (antibody)
RRID:AB_2620389
This polyclonal targets METTL16
View all literature mentionsHEK293T (cell line)
RRID:CVCL_0063
Cell line HEK293T is a Transformed cell line with a species of origin Homo sapiens (Human)
View all literature mentionsKasumi-1 (cell line)
RRID:CVCL_0589
Cell line Kasumi-1 is a Cancer cell line with a species of origin Homo sapiens (Human)
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RRID:CVCL_1609
Cell line NOMO-1 is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentionsNB4 (cell line)
RRID:CVCL_0005
Cell line NB4 is a Cancer cell line with a species of origin Homo sapiens (Human)
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NOD.Cg-Rag1tm1Mom Il2rg Tg(CMV-IL3.CSF2.KITLG)1Eav/J (organism)
RRID:IMSR_JAX:024099
Mus musculus with name NOD.Cg-Rag1tm1Mom Il2rg
B6.Cg-Tg(Mx1-cre)1Cgn/J (organism)
RRID:IMSR_JAX:003556
Mus musculus with name B6.Cg-Tg(Mx1-cre)1Cgn/J from IMSR.
View all literature mentions
B6.SJL-PtprcaPepc/BoyCrCrl (organism)
RRID:IMSR_CRL:564
Mus musculus with name B6.SJL-PtprcaPepc
OP9 (cell line)
RRID:CVCL_4398
Cell line OP9 is a Stromal cell line with a species of origin Mus musculus
View all literature mentionsMono-Mac-6 (cell line)
RRID:CVCL_1426
Cell line Mono-Mac-6 is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentionsML-2 (cell line)
RRID:CVCL_1418
Cell line ML-2 is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentionsMOLM-13 (cell line)
RRID:CVCL_2119
Cell line MOLM-13 is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentionsHL-60(TB) (cell line)
RRID:CVCL_A794
Cell line HL-60(TB) is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentionsTHP-1 (cell line)
RRID:CVCL_0006
Cell line THP-1 is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentions
NOD.Cg-Rag1tm1Mom Il2rg Tg(CMV-IL3.CSF2.KITLG)1Eav/J (organism)
RRID:IMSR_JAX:024099
Mus musculus with name NOD.Cg-Rag1tm1Mom Il2rg
