Sustained and intermittent hypoxia differentially modulate primary monocyte immunothrombotic responses to IL-1b stimulation

We replicated the intricate conditions of the venous valve pocket using a state-of-the-art hypoxia chamber with software-controlled oxygen cycling. We comprehensively studied the effects of sustained and intermitted hypoxia alone or in combination with IL-1b to mimic VTE-associated inflammatory stimuli on primary monocytes. Our transcriptome analysis revealed that sustained hypoxia limited the pro-inflammatory response induced by IL-1b, and triggered a metabolic shift in the monocyte transcriptome. On the other hand, intermittent hypoxia alone had a modest effect, but in combination with IL-1b it significant altered the expression of 2207 genes. Notably, intermittent hypoxia enhanced the IL-1b -stimulatory effects on several chemokine and interleukin genes (e.g., IL-19, IL-24, IL-32, MIF), as well as genes involved in coagulation (thrombomodulin) and fibrinolysis (VEGFA, MMP9, MMP14 and PAI-1. The RNA seq data are paired with validations on protein level in the published paper. Our findings provide valuable insights into how the different hypoxic profiles shape the immunothrombotic response of monocytes and shed new light on the early events in the pathogenesis of venous thrombosis. Setup of intermittent hypoxia combined with inflammatory IL-1b stimulation of primary monocytes. RNA isolated from nine donors was pooled into three pools of 3 donors each before sequencing. DGE and GSEA analyses were performed to map the effects of inflammation and hypoxia separately and combined. **RAW data not provided due to Norwegian laws on GDPR prevent us from publicly sharing the raw RNA sequences**