STAT1-dependent expression in kidney endothelium from mice challenged with LPS

Severe systemic inflammatory reactions, including sepsis, often lead to shock, organ failure and death, in part through an acute release of cytokines that promote vascular dysfunction. However, little is known about the vascular endothelial signaling pathways regulating the transcriptional profile in failing organs. Among all the cytokines altered during a cytokine storm, IL-6 levels are particularly informative, as they correlate with systemic disease severity and are highly predictive of mortality. Here, we show that loss of endothelial expression of the IL-6 pathway inhibitor, SOCS3, promotes a type I interferon (IFNI)-like gene signature in response to endotoxemia in mouse kidneys and brains. In cultured primary human endothelial cells, IL-6 induces a transient IFNI-like gene expression in a non-canonical, interferon-independent fashion. We further show that STAT3, which we had previously shown to control IL-6-driven endothelial barrier function, is dispensable for this activity. Instead, IL-6 promotes a transient increase in cytosolic mitochondrial DNA and requires STAT1, cGAS, STING, and the IRFs 1, 3, and 4. Inhibition of this pathway in endothelial-specific STING knockout mice or global STAT1 knockout mice leads to reduced severity of an acute endotoxemic challenge and prevents the endotoxin-induced IFNI-like gene signature. These results suggest that permeability and DNA sensing responses are driven by parallel pathways downstream of this cytokine, provide new insights into the complex response to acute inflammatory responses, and offer the possibility of novel therapeutic strategies for independently controlling the intracellular responses to IL-6 in order to tailor the inflammatory response. Overall design: STAT1KO (JAX #012606) or control C57Bl/6J (JAX #000664) were challenged with 250 ug of LPS and sacrificed 15 hours later. Whole organ RNA was oobtained from an aliquot of kidney single cell suspensions, while the reminder was processed for endoothelial cell enrichment prior to RNA isolation. Each experimental group includes two male and two female mice.