#5648 RIPK3 MEDIATES KIDNEY INJURY INDUCED BY A CYTOKINE STORM

Abstract Background and Aims Receptor-interacting protein kinase 3 (RIPK3) is traditionally involved in necroptosis, a regulated necrosis pathway which has been observed diseases associated to cell death inflammation. However, RIPK3 also inflammatory responses independent of necroptosis. In previous studies we demonstrated that deficiency does not prevent renal injury but protects from inflammation folic acid-induced AKI (FA-AKI), suggesting proinflammatory role independently death. the present work, aim explore kidney induced by cytokine storm. Method Lipopolysaccharide (LPS) was used induce storm-AKI mice. For this, female 12- 14-week-old wild type (WT), RIPK3-KO, MLKL-KO or NRLP3-KO C57BL/6J mice received single intraperitoneal (i.p) injection LPS 5 mg/kg vehicle were sacrificed 1h, 4h 24 hours later. To generate chimera mice, WT Ripk3-KO receptor irradiated deplete autologous bone marrow (BM). BM extracted femur tibia donor 107 cells transferred intravenous injection. After 1 month, LPS-AKI induced. Plasma collected assess function. Kidneys for RNA, studies, histologic studies. Additionally, group 1.65 necroptosis inhibitor Necrostatin-1 (Nec1) i.p. prior evaluate impact pathway. Liver, lungs heart systemic response. cultured murine immortalized tubular MCT cells, primary dendritic (BMDC) derived macrophages (BMDM) isolated studied. Cells stimulated with 100 ng/ml 6h RNA analyze response on supernatants LPS-stimulated BMDM BMDC stimulate cells. expression studied RT-PCR Western Blotting. Results The mRNA upregulated while Ripk3 improved survival function, less cytokines infiltrate. Necroptosis did seem be implicated storm-AKI, since neither Nec1 nor genetic MLKL ablation offered protection Systemic non-kidney organs milder deficiency. addition, inflammasome-related proteins this reduced Nlrp3-deficient developed after Next, explored BM-derived generating context, deficient exhibited better This supports mediates during storm-AKI. BMDMs mediated IL-6 Moreover, conditioned media LPS-exposed promoted partially ameliorated RIPK3-KO LPS-macrophage medium. Conclusion conclusion, storm These results identify as therapeutic target diseases.