Inhibition of Constitutively Activated Nuclear Factor-B Induces Reactive Oxygen Species- and Iron-Dependent Cell Death in Cutaneous T-Cell Lymphoma

Michael K. Kiessling ¹, Claus D. Klemke ², Marcin M. Kamiski ¹, Ioanna E. Galani ¹, Peter H. Krammer ¹, Karsten Gülow ^1*

¹Tumor Immunology Program, German Cancer Research Center (DFKZ), Heidelberg, Germany and ²Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Ruprecht Karl University of Heidelberg, Mannheim, Germany

^* To whom correspondence should be addressed. E-mail: k.guelow{at}dkfz-heidelberg.de.

	Abstract

Aberrant signaling of the nuclear facotr (NF-B) pathway has been identified as a mediator of survival and apoptosis resistance in leukemias and lymphomas. Here, we report that cell death of cutaneous T-cell lymphoma cell lines induced by inhibition of the NF-B pathway is independent of caspases or classic death receptors. We found that free intracellular iron and reactive oxygen species (ROS) are the main mediators of this cell death. Antioxidants such as N-Acetyl-L-cysteine and glutathione or the iron chelator desferrioxamine effectively block cell death in cutaneous T-cell lymphoma cell lines or primary T cells from Sézary patients. We show that inhibition of constitutively active NF-B causes down-regulation of ferritin heavy chain (FHC) that leads to an increase of free intracellular iron, which, in turn, induces massive generation of ROS. Furthermore, direct down-regulation of FHC by siRNA caused a ROS-dependent cell death. Finally, high concentrations of ROS induce cell death of malignant T cells. In contrast, T cells isolated from healthy donors do not display down-regulation of FHC and, therefore, do not show an increase in iron and cell death upon NF-B inhibition. In addition, in a murine T-cell lymphoma model, we show that inhibition of NF-B and subsequent down-regulation of FHC significantly delays tumor growth in vivo. Thus, our results promote FHC as a potential target for effective therapy in lymphomas with aberrant NF-B signaling. [Cancer Res 2009;69(6):2365–74]

Key Words: ferritin heavy chain, NF- B, reactive oxygen species