Necroptosis NOH HYUN JIN

Cell Death Apoptosis Normal Cell Autophagy Necrosis Normal Cell

Cell Death

As early as the mid-19th century, Rudolf Virchow taught that necrosis is a recognizable form of cell death; since then, pathologists have identified necrosis as both a cause and a consequence of disease. A century later, another form of cell death, apoptosis, was defined, and we now understand that this process is driven by a set of molecular mechanisms that “programs” the cell to die. It has often been assumed that necrosis is distinct from apoptosis, in part because of the belief that necrosis is not programmed by molecular events. It is now clear, however, that in some contexts, necrotic cell death can be driven by defined molecular pathways. N ENGL J MED 370;5 2014

Evolution of the Concept of Programmed Necrosis In some cell types, FAS can trigger non-apoptotic cell death that is independent of caspases but dependent on the adaptor protein FADD and the presence and enzymatic activity of the protein kinase RIP1. This milestone paper is the first report of RIP1‑dependent necroptosis. Nature Immunol, 200, 1:489-495, Tschopp’s group

Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury. Nature Chem. Biol. 1, 112–119 -- Degterev et al. identify Necrostatin-1 (Nec-1) as a potent inhibitor of TNF mediated necrosis and introduce the term “necroptosis” .

Identification of RIP3 as a modulator of Necroptosis 2009 2014 In contrast to RIP1, the loss of murine RIP3 results in no significant phenotypic defect, and has allowed the role of RIP3, and potentially necroptosis, to be examined in vivo during pathogenic infection and inflammatory disease models

Identification of MLKL as a modulator of Necroptosis 2012 2014 Identification of MLKL as a key modulator of necroptosis NSA NSA: necrosulfonamide

Molecular pathways for Necroptosis

TNFR1-mediated necroptosis : the prototype of regulated necrosis

Relationship between TRAF2 and TRADD - Cell line : HeLa(RIP3) - DNA plasmid: pEGF-c1, GFP-TRAF2, GFP-Peliino (each 1ug) - Transfection time: 24h, 48h - Transfection reagent: PEI GFP pEGF-c1 GFP-TRAF2 GFP-Pellino PEI (24h) PEI (48h) Actin TRADD

Relationship between TRAF2 and TRADD - Cell line : HeLa(RIP3) - DNA plasmid: pEGF-c1, GFP-TRAF2, GFP-Peliino (each 1ug) - Transfection time: 24h, 36h, 48h - Transfection reagent: PEI pEGF-c1 GFP-TRAF2 GFP-Pellino PEI (24h) PEI (48h) PEI (36h) HeLa (RIP3) GFP TRADD (short) TRADD (long) Actin (short) Actin (long)

What is RIP3-mediated MLKL activator under the ER stress condition induced by thapsigargin?

What is RIP3-mediated MLKL activator under the ER stress condition induced by thapsigargin?

What is RIP3-mediated MLKL activator under the ER stress condition induced by thapsigargin? - Thapsigargin (TG) condition : 9h / 1, 2 (uM) 1 2 - TG HeLa (RIP3) pMLKL RIP3 MLKL Actin RIP3 Actin CHOP 1 2 - TG HeLa (RIP3)

What is RIP3-mediated MLKL activator under the ER stress condition induced by thapsigargin? - Thapsigargin (TG) condition : 2uM / 0, 3h, 6h, 9h, 12h - 3 6 9 12 TG HeLa (RIP3) (h) HeLa (NC) pMLKL (long) CHOP MLKL pMLKL (short) Actin

What is RIP3-mediated MLKL activator under the ER stress condition induced by thapsigargin? - Thapsigargin (TG) condition : 12h / 0, 0.5, 1, 2, 4 (uM ) pMLKL (short) pMLKL (long) Actin (short) Actin (long) CHOP MLKL - 0.5 1 2 4 TG HeLa (RIP3) (uM)

What is RIP3-mediated MLKL activator under the glucose deprivation condition? 12 24 - GD HeLa (NC) HeLa (RIP3) pMLKL (short) pMLKL (long) Actin RIP3 RIP1 (short) RIP1 (long) pMLKL Actin RIP3 RIP1 - 12 24 GD HeLa (RIP3) HeLa (NC)