The interaction between vascular endothelial cells (ECs) and cancer cells is of vital importance to understand tumor dissemination. A paradigmatic cancer to study cell–cell interactions is classical Hodgkin Lymphoma (cHL) owing to its complex microenvironment. The role of the interplay between cHL and ECs remains poorly understood. Here we identify canonical WNT pathway activity as important for the mutual interactions between cHL cells and ECs. We demonstrate that local canonical WNT signaling activates cHL cell chemotaxis toward ECs, adhesion to EC layers and cell invasion using not only the Wnt-inhibitor Dickkopf, tankyrases and casein kinase 1 inhibitors but also knockdown of the lymphocyte enhancer binding-factor 1 (LEF-1) and β-catenin in cHL cells. Furthermore, LEF-1- and β-catenin-regulated cHL secretome promoted EC migration, sprouting and vascular tube formation involving vascular endothelial growth factor A (VEGF-A). Importantly, high VEGFA expression is associated with a worse overall survival of cHL patients. These findings strongly support the concept that WNTs might function as a regulator of lymphoma dissemination by affecting cHL cell chemotaxis and promoting EC behavior and thus angiogenesis through paracrine interactions.
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We thank Mrs S Schwoch for her technical assistance in the histological analysis of the CAM tumors and Mrs G Lutze for some advice in the vascular sprouting assay. This work was supported by grants of the Deutsche Forschungsgemeinschaft Ku 954/12-1 within the Forschergruppe FOR942. CEITEC—the Central European Institute of Technology is supported by CEITEC 2020 (LQ1601) project with financial contribution made by the Ministry of Education, Youths and Sports of the Czech Republic within special support paid from the National Programme for Sustainability II funds. VB and PJ are supported by the grant from the AZV CR, Ministry of Health, Czech Republic, NR 15-29793A. MMN is supported by the BMBF e:Bio Project MetastaSys (Ref.: 0316173) and TB/DK by BMBF e:Med Project MMML-Demonstrators (Ref.: 031A428B). SL is supported by the German Research Foundation grant SFB1002 TPC02.
The authors declare no conflict of interest.
FL, MH, SZ, and FvB did most of the experiments with AA, CD, SL, MMN and JW contributing to specific experiments, such as flow cytometry, Micro-CT analysis of the chick chorio-allantoic assay, time-lapse experiments, cell track analysis and data interpretation as well as chick chorio-allantoic model characterization. MS and WK performed IHC analysis. PJ and VB analyzed microarray data from Oncomine. MK and LT performed NMR studies and PO performed the corresponding cluster analysis. VB, JW, TB, FA and LT were involved in manuscript writing and final approval. FL and DK designed the research, analyzed and interpreted data and wrote the finally approved manuscript.
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Linke, F., Harenberg, M., Nietert, M. et al. Microenvironmental interactions between endothelial and lymphoma cells: a role for the canonical WNT pathway in Hodgkin lymphoma. Leukemia 31, 361–372 (2017). https://doi.org/10.1038/leu.2016.232
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