dsRNA mimics treatment of Melanoma and Human lymphatic cells

The crosstalk between cancer cells and the lymphatic vasculature has long been proposed to define competency for metastasis. Nevertheless, the discovery of selective blockers of lymphovascular niches has been compromised by the paucity of experimental systems for whole-body analyses of tumor progression. Spatio-temporal analyses in autochthonous melanomas and patient-derived xenografts identified double stranded RNA mimics (dsRNA nanoplexes) as potent repressors of lymphangiogenesis and metastasis. Mechanistically, dsRNA nanoplexes were found to suppress lymphangiogenic drivers in both tumor cells and their associated lymphatic vasculature (via MIDKINE and Vegfr3, respectively). This dual inhibitory action, driven by type I interferon, was not shared by FDA-approved antimelanoma treatments or by lymphangiogenic blockers in clinical testing. These results underscore the power of Vegfr3-lymphoreporters for pharmacological testing in otherwise aggressive cancers Overall design: Human Lymphatic Microvascular Endothelial Cells (HMVEC-dLy) (Lonza, MD) were grown in EGM™-2 medium supplemented with MV BulletKit (Lonza). SK-Mel-147 melanoma cells were grown in DMEN 10% FBS. 2.5 x 10^5 cells were seeded in MW6 plates 24h before treatment. When indicated, the cells were treated for 10h with 1 µg/ml BO-110 or vehicle control. Cells were then harvested, and total RNA was extracted. 500ng of total RNA samples were used. Each condition was analyzed in triplicates. Sequencing libraries were prepared with the "QuantSeq 3' mRNA-Seq Library Prep Kit (FWD) for Illumina" (Lexogen, Cat.No. 015) by following manufacturer instructions.