Oncostatin M induces epigenetic reprogramming in renal cell carcinoma-associated endothelial cells
Clear-cell renal cell carcinoma (ccRCC) is the most prevalent and lethal subtype of renal cell carcinoma. Genomic and epigenomic inactivation of the von Hippel-Lindau (VHL) tumor suppressor gene is a crucial early event in ccRCC pathogenesis (~75% of cases). Loss of VHL function leads to dysregulated hypoxia signaling, resulting in an extensive, disorganized, tortuous, and leaky vascular network within ccRCC tissue. It has been shown previously that Oncostatin M (OSM) secreted by the VHL-deficient kidney tubule cells can induce most of these ccRCC-associated vascular phenotypes which in turn promote tumor cell proliferation, immune cell infiltration, and metastasis. The influence of OSM to ECs could induce extensive and long-lasting transcriptomic changes in ECs although these ECs did not harbor any genomic mutations. Further studies indicate that OSM specifically activates endothelial cells (ECs) by inducing acetylation of histone 3 at lysine 14 (H3K14ac). Thus, we performed a variant of chromatin immunoprecipitation analysis, the Cleavage Under Targets and Tagmentation (CUT&TAG) sequencing, on OSM-treated and untreated ECs labeled with anti-H3K14ac antibody to validate the role of H3K14ac in the activation of ECs. Overall design: 4x104 HMEC-1 cells were seeded in 3.5 cm2 culture dish and grown overnight. The media was replaced with fresh complete media (Control; CT group), 10 ng/mL of rOSM (OSM group). Cells were harvested after 48 hr and performed CUT&TAG sequencing by Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme, TD903).