Michelle KelliherZelic, Matija2022-08-232022-08-232018-01-182018-01-3110.13028/M2GQ3Vhttps://hdl.handle.net/20.500.14038/32337Necroptosis, a type of regulated necrotic cell death, involves cell membrane permeabilization and has been implicated in various acute and chronic pro-inflammatory diseases, including ischemia-reperfusion injury and neurodegenerative diseases. By using in vitro reconstitution studies and a chemical inhibitor, the kinase activity of the serine/threonine kinase RIPK1 had been shown to regulate necroptotic signaling downstream of TNF and Toll-like receptors (TLRs). To investigate the contribution of RIPK1 kinase activity to inflammation and necroptosis in vivo, we generated kinase inactive RIPK1 knock-in mice. Utilizing fibroblasts and macrophages from these mice, we demonstrate that RIPK1 kinase activity is required for necroptotic complex formation and death induction downstream of TNFR1 and TLRs 3 and 4. We show that RIPK1 kinase inactive mice are resistant to TNF-induced shock and exhibit impaired upregulation of TNF-induced cytokines and chemokines in vitro and in vivo. By using bone marrow reconstitution experiments, we demonstrate that RIPK1 kinase activity in a non-hematopoietic lineage drives TNF-induced lethality. We establish that RIPK1 kinase activity is required for TNF-induced increases in intestinal and vascular permeability and clotting, and implicate endothelial cell necroptosis as an underlying factor contributing to TNF/zVAD-induced shock. Thus, work in this thesis reveals that RIPK1 kinase inhibitors may have promise in treating shock and sepsis.en-USLicensed under a Creative Commons licensehttp://creativecommons.org/licenses/by/4.0/Inflammationcell deathRIPK1TNFshocksepsisOther Immunology and Infectious DiseaseDoctoral Dissertationhttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1958&context=gsbs_diss&unstamped=1https://escholarship.umassmed.edu/gsbs_diss/95211459699gsbs_diss/9520000-0003-4901-5580