A unique death pathway keeps RIPK1 D325A mutant mice in check at embryonic day 10.5

Tumor necrosis factor receptor-1 (TNFR1) signaling, apart from its pleiotropic functions in inflammation, plays a role in embryogenesis as deficiency of varieties of its downstream molecules leads to embryonic lethality in mice. Caspase-8 noncleavable receptor interacting serine/threonine kinase 1 (RIPK1) mutations occur naturally in humans, and the corresponding D325A mutation in murine RIPK1 leads to death at early midgestation. It is known that both the demise ofRipk1D325A/D325Aembryos and the death ofCasp8−/−mice are initiated by TNFR1, but they are mediated by apoptosis and necroptosis, respectively. Here, we show that the defects inRipk1D325A/D325Aembryos occur at embryonic day 10.5 (E10.5), earlier than that caused byCasp8knockout. By analyzing a series of genetically mutated mice, we elucidated a mechanism that leads to the lethality ofRipk1D325A/D325Aembryos and compared it with that underliesCasp8deletion-mediated lethality. We revealed that the apoptosis inRipk1D325A/D325Aembryos requires a scaffold function of RIPK3 and enzymatically active caspase-8. Unexpectedly, caspase-1 and caspase-11 are downstream of activated caspase-8, and concurrent depletion ofCasp1andCasp11postpones the E10.5 lethality to embryonic day 13.5 (E13.5). Moreover, caspase-3 is an executioner of apoptosis at E10.5 inRipk1D325A/D325Amice as its deletion extends life ofRipk1D325A/D325Amice to embryonic day 11.5 (E11.5). Hence, an unexpected death pathway of TNFR1 controls RIPK1 D325A mutation-induced lethality at E10.5.