Death Receptor‐induced Necroptosis


Recent studies on cellular in vitro models have made it clear that death receptor (DR)‐induced necrosis, also called necroptosis, is a programmed form of necrotic cell death. In contrast, accidental necrosis is mainly induced by physicochemical stress, such as heath or chemicals. Many DRs signal to apoptosis or necroptosis depending on whether caspases are active or not. The kinase activities of receptor‐interacting protein 1 (RIP1) and RIP3 have emerged as crucial regulators of DR‐induced necroptosis. The precise execution mechanisms during necroptosis are still poorly understood but involve different cellular compartments such as the mitochondria, the lysosomes and the cell membrane. In vivo, necroptosis occurs mainly in pathophysiological processes such as ischaemia‐reperfusion injury in heart, brain and kidneys, viral infection and pancreatitis, and is capable of killing tumour cells that have developed strategies to evade apoptosis. Thus, detailed knowledge of necroptotic signalling may be exploited in therapeutic strategies.

Key concepts:

  • Necrosis is characterized by swelling of the endoplasmic reticulum, mitochondria and cytoplasm, with subsequent rupture of the plasma membrane and lysis of the cells.

  • Death receptor‐induced necrosis, also called necroptosis, is a programmed form of necrotic cell death. This is in contrast to necrosis induced by physicochemical stress, such as heath or hydrogen peroxide.

  • The necrosis‐signalling complex, or necrosome, consists of a complex containing FADD, caspase‐8, RIP1 and RIP3.

  • Expression of RIP3 renders cells permissive to death receptor‐induced necroptosis.

  • RIP1 and RIP3 kinase activity is crucial for necroptotic signalling.

  • Apoptosis counteracts necrotic signalling by cleavage of RIP1 and RIP3.

  • Necroptosis forms a back‐up cell death mode in case caspases activity is prevented or inhibited.

  • In vivo, necroptosis occurs mainly in pathophysiological processes such as ischaemia‐reperfusion injury in heart, brain and kidneys, viral infection and pancreatitis, and is capable of killing tumour cells that have developed strategies to evade apoptosis.

Keywords: necrosis–death receptors; TNF‐necrosome; receptor‐interacting kinase; mitochondria‐reactive oxygen species; metabolism

Figure 1.

Cell morphology of apoptotic and necrotic cells by transmission electron microscopy. (a) Unstimulated L929sAhFas fibrosarcoma cell. The cell shows microvilli protruding from the entire surface (arrowhead), a smoothly outlined nucleus with chromatin in the form of heterochromatin and well‐preserved cytoplasmic organelles. (b) Apoptotic L929sAhFas cell (treated with agonistic anti‐Fas for 1 h) with condensed and marginated (arrowhead) chromatin. Note the nucleolus (n) and damaged mitochondria (arrow). (c) Necrotic L929sAhFas cell (treated with TNF for 7 h) with clumps of chromatin with ill‐defined edges, swollen and completely disrupted mitochondria (arrow) and loss of plasma membrane integrity (arrowhead). Scale bars: 1 μm, N, nucleus. Pictures are courtesy of Katharina D'Herde and Dmitri Krysko (Ghent University).

Figure 2.

The necrosome, a new player in death receptor‐induced necroptosis. (a) Schematic representation of the different domains in human RIP1 and RIP3. We indicated the number of amino acids corresponding to each domain, the modifications important for NFκB activation (K63‐linked polyubiquitin chains on K377 in RIP1) or necroptosis (phosphorylation on RIP1S161 and RIP3 S199). The interaction between RIP1 and RIP3 is mediated by the RHIM domain. Abbreviations: KD, kinase domain; RHIM, RIP homotypic interaction motif; DD, death domain. (b) TNF‐induced necroptosis signalling complexes. Upon TNFR1 stimulation, TRADD provides, by binding RIP1, TRAF2 and cIAP1 and cIAP2, a scaffold for the assembly of complex I at the plasma membrane. This complex is crucial for activating NFκB and MAPK pathways. K63‐linked polyubiquitination of RIP1 by cIAPs results in interaction of RIP1 with the TAK1 (TGF (transforming growth factor)‐β‐activated kinase 1)/TAB2 (TAK‐1 binding protein 2)/3 complex. K63‐linked TRAF2 polyubiquitination can also recruit the TAK1/TAB2/3 complex. TAK1 activates the IKK complex, containing IKK‐α, IKK‐β and NEMO/IKKγ, which in turn phosphorylates IκB and results in its K48 polyubiquitination and proteasomal degradation. Once freed from its inhibitor, NFκB translocates to the nucleus and induces transcription. In a negative feedback loop, NFκB‐mediated upregulation of A20 and CYLD targets RIP1 for K63 deubiquitination, thereby abolishing its ability to activate NFκB. After receptor internalization TRADD‐dependent secondary cytosolic complexes are formed (complex II). The establishment of complex II involves FADD‐mediated recruitment and activation of caspase‐8, leading to RIP1 and RIP3 cleavage. In conditions where apoptosis is blocked by experimental (e.g. zVAD) or physiological conditions (e.g. viral inhibitors), RIP1 and RIP3 assemble a complex involving FADD and caspase‐8. The asterisk indicates that TRADD may also be part of this complex; however, this has not been formally proven. Mutual direct or indirect phosphorylation of RIP1 and RIP3 in the necrosome activates necroptotic signalling. In addition, TNF induces the formation of a TNFR1 signalling complex containing TRADD, RIP1, Nox1/NOXA1 and the small GTPase Rac1. Recruitment of Nox1 complex to TNFR1 is dependent on the adaptor proteins TRADD and RIP1 kinase. Upon activation of Nox1, superoxide anions and consecutive hydrogen peroxides are generated eventually contributing to necroptosis. See text for details.

Figure 3.

Different intracellular signalling events contribute to necroptosis. The kinase activity of RIP1 is needed to induce necroptosis induced by death receptors and several other stimuli. Necrotic cell death depends on the kinase activity of RIP1, which is inhibitable by the compound Nec‐1, and on a phosphorylation loop between RIP1 and RIP3. The main players in the propagation of necroptosis are calcium and mitochondria. Calcium controls the activation of PLA2 and calpains, inducing lipid peroxidation (LOOH) and cathepsin release from lysosomes, respectively Mitochondria contribute to necroptosis by excessive ROS production, mitochondrial permeability transition (mPT) or ATP depletion. RIP3 mediates enhanced glycolysis through glycogenolysis, and glutaminolysis, stimulating Krebs cycle and ROS production. See text for details. In addition, Bcl‐2 family members also contribute to necroptotic signalling, probably by affecting the mitochondrial function.



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Further Reading

Degterev A and Yuan J (2008) Expansion and evolution of cell death programmes. Nature Reviews. Molecular Cell Biology 9: 378–390.

Galluzzi L and Kroemer G (2008) Necroptosis: a specialized pathway of programmed necrosis. Cell 135: 1161–1163.

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Zong WX and Thompson CB (2006) Necrotic death as a cell fate. Genes & Development 20: 1–15.

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Declercq, Wim, Van Herreweghe, Franky, Berghe, Tom Vanden, and Vandenabeele, Peter(Dec 2009) Death Receptor‐induced Necroptosis. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0021566]