title: A genome-wide RNAi screen identifies proteins modulating aberrant FLT3-ITD signaling.
abstract: Fms-like tyrosine kinase-3 is a commonly mutated gene in acute myeloid leukemia, with about one-third of patients carrying an internal-tandem duplication of the juxtamembrane domain in the receptor (FLT3-ITD). FLT3-ITD exhibits altered signaling quality, including aberrant activation of STAT5. To identify genes affecting FLT3-ITD-mediated STAT5 signaling, we performed an esiRNA-based RNAi screen utilizing a STAT5-driven reporter assay. Knockdowns that caused reduced FLT3-ITD-mediated STAT5 signaling were enriched for genes encoding proteins involved in protein secretion and intracellular protein transport, indicating that modulation of protein transport processes could potentially be used to reduce constitutive STAT5 signaling in FLT3-ITD-positive cells. The relevance of KDELR1, a component involved in the Golgi-ER retrograde transport, was further analyzed. In FLT3-ITD-expressing leukemic MV4-11 cells, downregulation of KDELR1 resulted in reduced STAT5 activation, proliferation and colony-forming capacity. Stable shRNA-mediated depletion of KDELR1 in FLT3-ITD-expressing 32D cells likewise resulted in reduced STAT5 signaling and cell proliferation. Importantly, these cells also showed a reduced capacity to generate a leukemia-like disease in syngeneic C3H/HeJ mice. Together our data suggest intracellular protein transport as a potential target for FLT3-ITD driven leukemias, with KDELR1 emerging as a positive modulator of oncogenic FLT3-ITD activity.
authors.affiliation: Department of Medical Systems Biology, University Hospital and Medical Faculty Carl Gustav Carus, University of Technology Dresden, Dresden, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Caldarelli
authors.forename: J P
authors.initials: JP
authors.lastname: Müller
authors.forename: M
authors.initials: M
authors.lastname: Paskowski-Rogacz
authors.forename: K
authors.initials: K
authors.lastname: Herrmann
authors.forename: R
authors.initials: R
authors.lastname: Bauer
authors.forename: S
authors.initials: S
authors.lastname: Koch
authors.forename: A K
authors.initials: AK
authors.lastname: Heninger
authors.forename: D
authors.initials: D
authors.lastname: Krastev
authors.forename: L
authors.initials: L
authors.lastname: Ding
authors.forename: S
authors.initials: S
authors.lastname: Kasper
authors.forename: T
authors.initials: T
authors.lastname: Fischer
authors.forename: M
authors.initials: M
authors.lastname: Brodhun
authors.forename: F-D
authors.initials: FD
authors.lastname: Böhmer
authors.forename: F
authors.initials: F
authors.lastname: Buchholz
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D016678:Genome; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D011506:Proteins / Q000502:physiology*; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D050799:STAT5 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D051941:fms-Like Tyrosine Kinase 3 / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D011506:Proteins; D050799:STAT5 Transcription Factor; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3