RAGE interacts with the necroptotic protein RIPK3 and mediates transfusion‐induced danger signal release

Abstract

RBC transfusion is associated with increased morbidity and mortality in critically ill patients. Endothelial cell necroptosis and subsequent damage-associated molecular pattern (DAMP) release has been identified as a mechanism of injury following RBC transfusion. Mounting evidence implicates the pro-inflammatory pattern recognition receptor, Receptor for Advanced Glycation End Products (RAGE), in initiating cell death programmes such as necroptosis. Here, we demonstrate the role of RAGE in endothelial necroptosis, as deletion of RAGE attenuates necroptotic cell death in response to TNFa, LPS or CpG-DNA. We show direct interaction of RAGE with the critical mediator of necroptosis, Receptor Interacting Protein Kinase 3 (RIPK3), during necroptosis. Furthermore, we observe decreased plasma High Mobility Group Box 1 (HMGB1) and RIPK3 levels in RAGE deficient mice compared to WT mice post-transfusion, substantiating the role for RAGE in transfusion-induced DAMP release in vivo. Collectively, these findings underscore RAGE as an essential mediator of regulated necrosis and post-transfusion DAMP release. Further studies to understand the role of RAGE and the necroptotic pathway in transfusion-induced organ injury may offer key targets to mitigate transfusion-related risks, including the risk of ARDS, in susceptible hosts.

MLKL Regulates Rapid Cell Death-independent HMGB1 Release in RSV Infected Airway Epithelial Cells

“…Moreover, in this setting, the induced phosphorylation of RIPK1 and RIPK3 was attenuated in the presence of the RAGE antagonist, supporting the notion that activation of RAGE in particular microenvironments can promote necroptosis. Of note, RAGE has been shown to directly interact with RIPK3 in endothelial cells ( Faust et al, 2020 ), supporting our findings. However, we cannot exclude the possibility that the HMGB1/RAGE pathway induces a downstream mediator, such as TNF-a, that then initiates necroptosis.…”

“…Necroptosis can be induced through the activation of various pattern recognition receptors, including RAGE ( Upton et al, 2012 ; Gao et al, 2014 ; Schock et al, 2017 ; Amarante-Mendes et al, 2018 ; Faust et al, 2020 ). As HMGB1 is a ligand of RAGE, we hypothesised that the early release of HMGB1 activates RAGE, triggering necroptosis and late HMGB1 release.…”

Novel, non-conventional pathways of necroptosis in the heart and other organs: Molecular mechanisms, regulation and inter-organelle interplay

“…Finally, receptor for advanced glycation end products (RAGE) has been identified as another non-conventional mediator of necroptosis in a murine model of red blood cell transfusion. RAGE knockout in mice has been found to attenuate necrosis-like cell death as evidenced by decreased plasma levels of RIPK3 and HMGB1 [138]. Further, in vitro investigations have revealed that knockdown of RAGE suppressed necroptosis of human lung endothelial cells while RAGE was found to directly interact with RIPK3 in both the nucleus and cytosol [138] supporting the relevance of this novel identified mediator of necroptosis signalling.…”

Neuronal DAMPs exacerbate neurodegeneration via astrocytic RIPK3 signaling

“…Future work will be needed to define the signaling events that mediate RAGEdependent RIPK3 activation. A recent study demonstrated co-immunoprecipitation of RIPK3 with RAGE in an endothelial cell line following stimulation with TNF-a 78 , but the nature of this interaction and whether it happens under natural conditions in vivo remains to be established.…”