title: NOX4-driven ROS formation mediates PTP inactivation and cell transformation in FLT3ITD-positive AML cells.
abstract: Activating mutations of FMS-like tyrosine kinase 3 (FLT3), notably internal tandem duplications (ITDs), are associated with a grave prognosis in acute myeloid leukemia (AML). Transforming FLT3ITD signal transduction causes formation of reactive oxygen species (ROS) and inactivation of the protein-tyrosine phosphatase (PTP) DEP-1/PTPRJ, a negative regulator of FLT3 signaling. Here we addressed the underlying mechanisms and biological consequences. NADPH oxidase 4 (NOX4) messenger RNA and protein expression was found to be elevated in FLT3ITD-positive cells and to depend on FLT3ITD signaling and STAT5-mediated activation of the NOX4 promoter. NOX4 knockdown reduced ROS levels, restored DEP-1 PTP activity and attenuated FLT3ITD-driven transformation. Moreover, Nox4 knockout (Nox4(-/-)) murine hematopoietic progenitor cells were refractory to FLT3ITD-mediated transformation in vitro. Development of a myeloproliferative-like disease (MPD) caused by FLT3ITD-transformed 32D cells in C3H/HeJ mice, and of a leukemia-like disease in mice transplanted with MLL-AF9/ FLT3ITD-transformed murine hematopoietic stem cells were strongly attenuated by NOX4 downregulation. NOX4-targeting compounds were found to counteract proliferation of FLT3ITD-positive AML blasts and MPD development in mice. These findings reveal a previously unrecognized mechanism of oncoprotein-driven PTP oxidation, and suggest that interference with FLT3ITD-STAT5-NOX4-mediated overproduction of ROS and PTP inactivation may have therapeutic potential in a subset of AML.
authors: Institute of Molecular Cell Biology, CMB, Jena University Hospital, Jena, Germany.
authors: A K
authors: AK
authors: Jayavelu
authors: Institute of Molecular Cell Biology, CMB, Jena University Hospital, Jena, Germany.
authors: J P
authors: JP
authors: Müller
authors: Institute of Molecular Cell Biology, CMB, Jena University Hospital, Jena, Germany.
authors: R
authors: R
authors: Bauer
authors: Institute of Molecular Cell Biology, CMB, Jena University Hospital, Jena, Germany.
authors: S-A
authors: SA
authors: Böhmer
authors: Institute of Molecular Cell Biology, CMB, Jena University Hospital, Jena, Germany.
authors: J
authors: J
authors: Lässig
authors: Division of Hematology and Hemostaseology and Ludwig Boltzmann Cluster Oncology, Department of Internal Medicine I, Medical University of Vienna, Währinger Gürtel 18-20, Vienna, Austria.
authors: S
authors: S
authors: Cerny-Reiterer
authors: Division of Hematology and Hemostaseology and Ludwig Boltzmann Cluster Oncology, Department of Internal Medicine I, Medical University of Vienna, Währinger Gürtel 18-20, Vienna, Austria.
authors: W R
authors: WR
authors: Sperr
authors: Division of Hematology and Hemostaseology and Ludwig Boltzmann Cluster Oncology, Department of Internal Medicine I, Medical University of Vienna, Währinger Gürtel 18-20, Vienna, Austria.
authors: P
authors: P
authors: Valent
authors: Ludwig Boltzmann Institute for Cancer Research, Institute of Animal Breeding and Genetics, University of Veterinary Medicine Vienna, Veterinärplatz 1, Vienna, Austria, and Medical University of Vienna, Vienna, Austria.
authors: B
authors: B
authors: Maurer
authors: Ludwig Boltzmann Institute for Cancer Research, Institute of Animal Breeding and Genetics, University of Veterinary Medicine Vienna, Veterinärplatz 1, Vienna, Austria, and Medical University of Vienna, Vienna, Austria.
authors: R
authors: http://orcid.org/0000-0003-0918-9463
authors: R
authors: Moriggl
authors: Institut für Kardiovaskuläre Physiologie, Goethe-Universität Frankfurt, Sandhofstrasse 2, Frankfurt, Germany.
authors: K
authors: K
authors: Schröder
authors: Cardiovascular Division, King's College London British Heart Foundation Centre, 125 Coldharbour Lane, London, UK.
authors: A M
authors: AM
authors: Shah
authors: Department of Hematology/Oncology, Jena University Hospital, Erlanger Allee 101, Jena, Germany.
authors: M
authors: M
authors: Fischer
authors: Department of Hematology/Oncology, Jena University Hospital, Erlanger Allee 101, Jena, Germany.
authors: S
authors: S
authors: Scholl
authors: Department of Medicine II, Hematology/Oncology, Goethe University, Theodor-Stern-Kai 7, Frankfurt, Germany.|German Cancer Consortium (DKTK), Heidelberg, Germany.|German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors: J
authors: J
authors: Barth
authors: Department of Medicine II, Hematology/Oncology, Goethe University, Theodor-Stern-Kai 7, Frankfurt, Germany.
authors: T
authors: T
authors: Oellerich
authors: Department of Medicine II, Hematology/Oncology, Goethe University, Theodor-Stern-Kai 7, Frankfurt, Germany.
authors: T
authors: T
authors: Berg
authors: Department of Medicine II, Hematology/Oncology, Goethe University, Theodor-Stern-Kai 7, Frankfurt, Germany.
authors: H
authors: H
authors: Serve
authors: Clinic for Hematology and Oncology, University Hospital Magdeburg, Leipziger Str. 44, Magdeburg, Germany.
authors: S
authors: S
authors: Frey
authors: Clinic for Hematology and Oncology, University Hospital Magdeburg, Leipziger Str. 44, Magdeburg, Germany.
authors: T
authors: T
authors: Fischer
authors: Clinic for Hematology and Oncology, University Hospital Magdeburg, Leipziger Str. 44, Magdeburg, Germany.
authors: F H
authors: FH
authors: Heidel
authors: Institute of Molecular Cell Biology, CMB, Jena University Hospital, Jena, Germany.
authors: F-D
authors: FD
authors: Böhmer
mesh_terms: D000818:Animals; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000473:pathology*; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000235:genetics / Q000502:physiology*; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D054631:Receptor-Like Protein Tyrosine Phosphatases, Class 3 / Q000032:analysis; D020080:Tandem Repeat Sequences; D051941:fms-Like Tyrosine Kinase 3 / Q000032:analysis / Q000502:physiology*
chemical_list: D017382:Reactive Oxygen Species; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; C498910:NOX4 protein, human; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3; C486288:PTPRJ protein, human; D017027:Protein Tyrosine Phosphatases; D054631:Receptor-Like Protein Tyrosine Phosphatases, Class 3