Classical Hodgkin lymphoma (cHL) has a typical clinical manifestation, with dissemination involving functionally neighboring lymph nodes. The factors involved in the spread of lymphoma cells are poorly understood. Here we show that cHL cell lines migrate with higher rates compared with non-Hodgkin lymphoma cell lines. cHL cell migration, invasion and adhesion depend on autocrine WNT signaling as revealed by the inhibition of WNT secretion with the porcupine inhibitors Wnt-C59/IWP-2, but did not affect cell proliferation. While application of recombinant WNT5A or WNT5A overexpression stimulates HL cell migration, neither WNT10A, WNT10B nor WNT16 did so. Time-lapse studies revealed an amoeboid type of cell migration modulated by WNT5A. Reduced migration distances and velocity of cHL cells, as well as altered movement patterns, were observed using porcupine inhibitor or WNT5A antagonist. Knockdown of Frizzled5 and Dishevelled3 disrupted the WNT5A-mediated RHOA activation and cell migration. Overexpression of DVL3-K435M or inhibition of ROCK (Rho-associated protein kinase) by Y-27632/H1152P disrupted cHL cell migration. In addition to these mechanistic insights into the role of WNT5A in vitro, global gene expression data revealed an increased WNT5A expression in primary HL cells in comparison with normal B-cell subsets and other lymphomas. Furthermore, the activity of both porcupine and WNT5A in cHL cells had an impact on lymphoma development in the chick chorionallantoic membrane assay. Massive bleeding of these lymphomas was significantly reduced after inhibition of WNT secretion by Wnt-C59. Therefore, a model is proposed where WNT signaling has an important role in regulating tumor-promoting processes.
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We thank Moritz Harenberg for evaluating the vascularization of the CAM tumors in a blinded manner. We are very thankful to Mrs S Schwoch, Mrs C Zelent and Mrs S Hellbach for their technical assistance in the histological analysis of the CAM tumors. We kindly acknowledge Dr Aldo Ferrari from the ETH Zürich to provide the sketch on the amoeboid migration type (integrated into Figure 3a). This work was supported by grants of the Deutsche Forschungsgemeinschaft Ku 954/12-1 within the Forschergruppe FOR942.
FL, SZ, FvB did most of the experiments with CD, SL, MMN and JW contributing to specific experiments as Micro-CT analysis of the chick chorioallantoic assay, time-lapse experiments, cell track analysis and data interpretation, as well as chick chorioallantoic model characterization. PJ and VB analyzed microarray data from Oncomine. JW, TB, WK, FA, TP and LT were involved in manuscript writing and the final approval. VB designed experiments and wrote the manuscript. FL and DK designed the research, analyzed, interpreted data and wrote the manuscript.
The authors declare no conflict of interest.
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Linke, F., Zaunig, S., Nietert, M. et al. WNT5A: a motility-promoting factor in Hodgkin lymphoma. Oncogene 36, 13–23 (2017). https://doi.org/10.1038/onc.2016.183
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