title: A noncanonical Flt3ITD/NF-κB signaling pathway represses DAPK1 in acute myeloid leukemia.
abstract: OBJECTIVE
  Death-associated protein kinase 1 (DAPK1), a tumor suppressor, is a rate-limiting effector in an endoplasmic reticulum (ER) stress-dependent apoptotic pathway. Its expression is epigenetically suppressed in several tumors. A mechanistic basis for epigenetic/transcriptional repression of DAPK1 was investigated in certain forms of acute myeloid leukemia (AML) with poor prognosis, which lacked ER stress-induced apoptosis.
  
  
  METHODS
  Heterogeneous primary AMLs were screened to identify a subgroup with Flt3ITD in which repression of DAPK1, among NF-κB-and c-Jun-responsive genes, was studied. RNA interference knockdown studies were carried out in an Flt3ITD(+) cell line, MV-4-11, to establish genetic epistasis in the pathway Flt3ITD-TAK1-DAPK1 repression, and chromatin immunoprecipitations were carried out to identify proximate effector proteins, including TAK1-activated p52NF-κB, at the DAPK1 locus.
  
  
  RESULTS
  AMLs characterized by normal karyotype with Flt3ITD were found to have 10- to 100-fold lower DAPK1 transcripts normalized to the expression of c-Jun, a transcriptional activator of DAPK1, as compared with a heterogeneous cytogenetic category. In addition, Meis1, a c-Jun-responsive adverse AML prognostic gene signature was measured as control. These Flt3ITD(+) AMLs overexpress relB, a transcriptional repressor, which forms active heterodimers with p52NF-κB. Chromatin immunoprecipitation assays identified p52NF-κB binding to the DAPK1 promoter together with histone deacetylase 2 (HDAC2) and HDAC6 in the Flt3ITD(+) human AML cell line MV-4-11. Knockdown of p52NF-κB or its upstream regulator, NF-κB-inducing kinase (NIK), de-repressed DAPK1. DAPK1-repressed primary Flt3ITD(+) AMLs had selective nuclear activation of p52NF-κB.
  
  
  CONCLUSIONS
  Flt3ITD promotes a noncanonical pathway via TAK1 and p52NF-κB to suppress DAPK1 in association with HDACs, which explains DAPK1 repression in Flt3ITD(+) AML.
authors.affiliation: Indiana University Melvin and Bren Simon Cancer Center, Departments of Medicine (Hematology/Oncology Division), Indiana University School of Medicine, Indianapolis, IN 46202.|Veterans Affairs Medical Center, Indianapolis, IN 46202.
authors.forename: Rajasubramaniam
authors.initials: R
authors.lastname: Shanmugam
authors.affiliation: Department of Microbiology and Immunology, Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD.
authors.forename: Padmaja
authors.initials: P
authors.lastname: Gade
authors.affiliation: Indiana University Melvin and Bren Simon Cancer Center, Departments of Medicine (Hematology/Oncology Division), Indiana University School of Medicine, Indianapolis, IN 46202.|Veterans Affairs Medical Center, Indianapolis, IN 46202.
authors.forename: Annique
authors.initials: A
authors.lastname: Wilson-Weekes
authors.affiliation: Indiana University Melvin and Bren Simon Cancer Center, Departments of Medicine (Hematology/Oncology Division), Indiana University School of Medicine, Indianapolis, IN 46202.
authors.forename: Hamid
authors.initials: H
authors.lastname: Sayar
authors.affiliation: Indiana University Melvin and Bren Simon Cancer Center, Departments of Medicine (Hematology/Oncology Division), Indiana University School of Medicine, Indianapolis, IN 46202.|Veterans Affairs Medical Center, Indianapolis, IN 46202.
authors.forename: Attaya
authors.initials: A
authors.lastname: Suvannasankha
authors.affiliation: Biostatistics and Computational Biology, Indiana University School of Medicine, Indianapolis, IN 46202.
authors.forename: Chirayu
authors.initials: C
authors.lastname: Goswami
authors.affiliation: Biostatistics and Computational Biology, Indiana University School of Medicine, Indianapolis, IN 46202.
authors.forename: Lang
authors.initials: L
authors.lastname: Li
authors.affiliation: Indiana University Melvin and Bren Simon Cancer Center, Departments of Medicine (Hematology/Oncology Division), Indiana University School of Medicine, Indianapolis, IN 46202.
authors.forename: Sushil
authors.initials: S
authors.lastname: Gupta
authors.affiliation: Indiana University Melvin and Bren Simon Cancer Center, Departments of Medicine (Hematology/Oncology Division), Indiana University School of Medicine, Indianapolis, IN 46202.
authors.forename: Angelo A
authors.initials: AA
authors.lastname: Cardoso
authors.affiliation: Indiana University Melvin and Bren Simon Cancer Center, Departments of Medicine (Hematology/Oncology Division), Indiana University School of Medicine, Indianapolis, IN 46202.
authors.forename: Tareq Al
authors.initials: TA
authors.lastname: Baghdadi
authors.affiliation: Indiana University Health Systems, Indianapolis, IN.
authors.forename: Katie J
authors.initials: KJ
authors.lastname: Sargent
authors.affiliation: Indiana University Melvin and Bren Simon Cancer Center, Departments of Medicine (Hematology/Oncology Division), Indiana University School of Medicine, Indianapolis, IN 46202.
authors.forename: Larry D
authors.initials: LD
authors.lastname: Cripe
authors.affiliation: Department of Microbiology and Immunology, Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD.
authors.forename: Dhananjaya V
authors.initials: DV
authors.lastname: Kalvakolanu
authors.affiliation: Indiana University Melvin and Bren Simon Cancer Center, Departments of Medicine (Hematology/Oncology Division), Indiana University School of Medicine, Indianapolis, IN 46202.|Veterans Affairs Medical Center, Indianapolis, IN 46202.
authors.forename: H Scott
authors.initials: HS
authors.lastname: Boswell
mesh_terms: D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D064550:Death-Associated Protein Kinases; D044127:Epigenesis, Genetic; D020869:Gene Expression Profiling; D015973:Gene Expression Regulation, Leukemic; D018922:HL-60 Cells; D006801:Humans; D059785:Karyotype; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000378:metabolism*; D020930:MAP Kinase Kinase Kinases / Q000378:metabolism; D009154:Mutation; D052003:NF-kappa B p52 Subunit / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D015398:Signal Transduction; D020080:Tandem Repeat Sequences; D014407:Tumor Cells, Cultured; D051941:fms-Like Tyrosine Kinase 3 / Q000235:genetics / Q000378:metabolism*; D000099151:MAP Kinase Kinase Kinase 7
chemical_list: D051017:Apoptosis Regulatory Proteins; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D064550:Death-Associated Protein Kinases; D020930:MAP Kinase Kinase Kinases; D052003:NF-kappa B p52 Subunit; D051941:fms-Like Tyrosine Kinase 3; D000099151:MAP Kinase Kinase Kinase 7; C497970:FLT3 protein, human; C579096:DAPK1 protein, human
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