#4234 adam17 modulates ifn-mediated inflammation in renal tubular cells

Abstract Background and Aims In diabetic patients, hyperglycemia has been associated with a hypoxic state that contributes to an increased rate of morbidity due to Acute Kidney Failure (AKI). For this reason, we are interested in studying the molecular mechanisms involved in the development of AKI to contribute to the search for effective therapies that can help prevent these episodes. We focused on ADAM17, a disintegrin and metalloproteinase that sheds the ectodomains of cell surface proteins, among which inflammatory cytokines stand out. In an animal model, we showed that the absence of ADAM17 in the renal tubule reduces protein expression related to fibrosis and inflammation induced by diabetes. In this context, we aimed to study the role of ADAM17 in modifying the expression of inflammatory molecules regulated by interferon (IFN) in renal proximal tubule cells subjected to hyperglycemic stress. In this study, we analyzed the effect of ADAM17 deletion in human renal tubular cells (HKC-8) incubated in high-glucose medium and subjected to hypoxia. Method ADAM17 was depleted using CRISPR/Cas9 (ADAM17-KO). Cells were incubated for 6 h or 24 h with high (35 mM HG) or normal (5.5 mM NG) glucose concentrations and were subjected to hypoxia plus 2h re-oxygenation (HIPX). Gene expression of several IFN-regulated inflammatory markers (IL6, IL8, CXCL10, ISG15, IL1a, IL1b, IL12A, CCL5, and CCL2) was determined in wild-type (WT) and ADAM17-KO cells. Results The absence of ADAM17 in HKC-8 cells induced changes in the mRNA expression of genes controlled by IFN, which is accentuated by the hyperglycemic stimulus. Under hypoxic and re-oxygenation conditions, these changes were in the same line, with more discrete values compared to control cells. TNFa, CXCL10 and LNC2 were the most relevant genes reduced in injury conditions in the ADAM17-KO cell line. Conclusion ADAM17 deletion modulates the expression of most IFN-regulated genes that could undergo post-transcriptional changes and subsequently act as an acute mechanism of protection from injury. The results show that ADAM17 plays a major role in the underlying inflammatory response to acute kidney injury, indicating its potential as a therapeutic target. The analysis of circulating proteins will provide information about this process to better understand the mechanism of action of ADAM17 in the protection of renal tubular damage.