Fibroblasts in Nodular Sclerosing Classical Hodgkin Lymphoma Are Defined by a Specific Phenotype and Protect Tumor Cells From Brentuximab-Vedotin Induced Injury. Fibroblasts in Nodular Sclerosing Classical Hodgkin Lymphoma Are Defined by a Specific Phenotype and Protect Tumor Cells From Brentuximab-Vedotin Induced Injury

Several studies have described a crosstalk between the tumour cells of cHL, the Hodgkin- and Reed-Sternberg (HRS) cells, and cancer-associated fibroblasts (CAF). However, to date a deep molecular characterization of these fibroblasts is lacking. Aim of the present study therefore was a comprehensive characterization of these fibroblasts. Overall design: Gene expression of fibroblasts isolated from primary lymph node suspensions (LA = lymphoadenitis, n=7, cHL_MC = classical Hodgkin lymphoma mixed cellularity subtype, n=5; cHL_NS = classical Hodgkin lymphoma nodular sclerosing subtype, n=8; Fib = primary fibroblasts culture at passage 4-6 in vitro, KB = arbitrary sample ID prefix, FST = arbitrary sample ID prefix). As previously shown for other types of cancer-associated fibroblasts, treatment by luteolin could reverse this fibroblast phenotype. Working concentration of 30 µM luteolin was added to sample KB33 and harvested 48 h post treatment for comparative analysis with Fib_cHL_NS samples. Three biological replicates were applied. The cHL specific adherence-based interaction of both cell fractions revealed an initiation of inflammatory key regulators. The classical Hodgkin lymphoma cell line L-428 and adherent activated fibroblasts cells from primary nodular sclerosis cHL were used. Floating cell fractions were removed by repeated washing. Adherent interacting coculture layer was harvested for comparative gene expression analysis to mock coculture. Mock coculture was set up separated in equivalent ratios and pooled before RNA analysis. Three biological replicates were applied.