High intracellular expression of death receptor TRAIL-R2 correlates with poor prognosis for different tumor entities and thus suggests tumor-promoting activity of intracellular TRAIL-R2. We demonstrate that TRAIL-R2 interacts with the core Microprocessor components Drosha and DGCR8 and the associated regulatory proteins p68, hnRNPA1, NF45 and NF90 in the nucleus. Knockdown of TRAIL-R2 enhances Drosha-mediated processing of pri-let-7 resulting in increased levels of mature let-7, reduced expression of let-7-targets Lin28B and HMGA2 and inhibition of cell proliferation. In contrast, high abundance of nuclear TRAIL-R2, often detected in pancreatic cancer, correlates with enhanced expression of HMGA2 and dictates worse prognosis. Importantly, knockdown of TRAIL-R2 inhibits pancreatic tumor growth in an orthotopic xenotransplantation mouse model and reduced nuclear levels of TRAIL-R2 accompany differentiation of pancreatic epithelial cells in vitro. In conclusion, we define a novel function of nuclear TRAIL death receptor contributing to malignancy by inhibition of let-7-maturation (Haselmann et al., Gastroenterology epub ahead of print).

In extension to our work on pancreatic cancer we further show nuclear TRAIL-R2 functions to be of relevance in breast cancer bone metastasis. Stably shRNA- transfected clones of MDAMB231 cells revealed metastatic lesions in only 2/12 mice upon TRAIL-R2 knock-down, whereas TRAIL-R1 and control knock-down clones exhibited multiple metastases throughout the groups of 12 mice each. Under in vitro conditions some decreased apoptosis rate was observed in both TRAIL-R knock-down clonal populations compared to the controls upon TRAIL treatment. Preliminary results suggest Mesenchymal-Epithelial-Transition (as indicated by increased E-Cadherin expression in TRAIL-R2 knock-down cells) as a mechanism for reduced metastasis.

In summary, we show nuclear death receptor TRAIL-R2 to significantly contribute to malignant progression in two different pre-clinical tumor models. Thus, targeted intervention to prevent nuclear localization may serve as a novel therapeutic strategy.

Supported by DFG (TR 1063/2-1 and TR 1063/3-1 - SKELMET FOR 1586).

Citation Format: Holger Kalthoff, Verena Haselmann, Alexandra Kurz, Uwe Bertsch, Sebastian Huebner, Hendrik Fritsche, Charlotte Hauser, Christian Schem, Rob Tower, Thorsten Heilmann, Sanjay Tiwari, Claus C. Glüer, Anna Trauzold. Trail-R2: A death receptor turns malignant upon nuclear localization. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2955. doi:10.1158/1538-7445.AM2014-2955