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.
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We thank Drs Jack Arbiser, Carmen Döbele, Adrian Manea, Siavosh Mahboobi, Frank Schnütgen and Maya Simionescu for the kind provision of reagents. We are grateful for support by the Deutsche Akademische Austauschdienst (to AKJ), by the Deutsche Forschungsgemeinschaft to F-DB (BO 1043/10), TF and FHH (FI405/5-1), to KS (SFB815/TP1 and SCHR1241/1-1), and to AKJ (associated member of GRK 1715), by the German Jose-Carreras Leukemia Society (DJCLS SP12/08) to FHH, by the Austrian Science Fund (FWF) SFB project F4704 and F4707 to RM and PV, by the LOEWE Center for Cell and Gene Therapy by the German Cancer Consortium (DKTK) to TB and TO, the Fraunhofer Gesellschaft (graduate school translational research innovation pharma, TRIP) to KS, and the German Center for Cardiovascular Research (DZHK) to KS.
The authors declare no conflict of interest.
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Jayavelu, A., Müller, J., Bauer, R. et al. NOX4-driven ROS formation mediates PTP inactivation and cell transformation in FLT3ITD-positive AML cells. Leukemia 30, 473–483 (2016). https://doi.org/10.1038/leu.2015.234
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