title: KIT with D816 mutations cooperates with CBFB-MYH11 for leukemogenesis in mice.
abstract: KIT mutations are the most common secondary mutations in inv(16) acute myeloid leukemia (AML) patients and are associated with poor prognosis. It is therefore important to verify that KIT mutations cooperate with CBFB-MYH11, the fusion gene generated by inv(16), for leukemogenesis. Here, we transduced wild-type and conditional Cbfb-MYH11 knockin (KI) mouse bone marrow (BM) cells with KIT D816V/Y mutations. KIT transduction caused massive BM Lin(-) cell death and fewer colonies in culture that were less severe in the KI cells. D816Y KIT but not wild-type KIT enhanced proliferation in Lin(-) cells and led to more mixed lineage colonies from transduced KI BM cells. Importantly, 60% and 80% of mice transplanted with KI BM cells expressing D816V or D816Y KIT, respectively, died from leukemia within 9 months, whereas no control mice died. Results from limiting dilution transplantations indicate higher frequencies of leukemia-initiating cells in the leukemia expressing mutated KIT. Signaling pathway analysis revealed that p44/42 MAPK and Stat3, but not AKT and Stat5, were strongly phosphorylated in the leukemia cells. Finally, leukemia cells carrying KIT D816 mutations were sensitive to the kinase inhibitor PKC412. Our data provide clear evidence for cooperation between mutated KIT and CBFB-MYH11 during leukemogenesis.
authors.affiliation: Oncogenesis and Development Section, National Human Genome Research Institute, National Institutes of Health (NIH), 49 Convent Drive, Bethesda, MD 20892, USA.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhao
authors.forename: Jan J
authors.initials: JJ
authors.lastname: Melenhorst
authors.forename: Lemlem
authors.initials: L
authors.lastname: Alemu
authors.forename: Martha
authors.initials: M
authors.lastname: Kirby
authors.forename: Stacie
authors.initials: S
authors.lastname: Anderson
authors.forename: Maggie
authors.initials: M
authors.lastname: Kench
authors.forename: Shelley
authors.initials: S
authors.lastname: Hoogstraten-Miller
authors.forename: Lauren
authors.initials: L
authors.lastname: Brinster
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Kamikubo
authors.forename: D Gary
authors.initials: DG
authors.lastname: Gilliland
authors.forename: P Paul
authors.initials: PP
authors.lastname: Liu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016026:Bone Marrow Transplantation; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018450:Disease Progression; D005260:Female; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007938:Leukemia / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009154:Mutation; D015514:Oncogene Proteins, Fusion / Q000235:genetics* / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics* / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism; D050799:STAT5 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction; D019311:Staurosporine / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: C095514:CBFbeta-MYH11 fusion protein; D015514:Oncogene Proteins, Fusion; D050796:STAT3 Transcription Factor; D050799:STAT5 Transcription Factor; D049452:Green Fluorescent Proteins; D019009:Proto-Oncogene Proteins c-kit; D051057:Proto-Oncogene Proteins c-akt; D020928:Mitogen-Activated Protein Kinases; D019311:Staurosporine; C059539:midostaurin