NINJ1 mediates plasma membrane rupture during lytic cell death

Plasma membrane rupture (PMR) is the final cataclysmic event in lytic cell death. PMR releases intracellular molecules known as damage-associated molecular patterns (DAMPs) that propagate the inflammatory response(1-3). The underlying mechanism of PMR, however, is unknown. Here we show that the cell-surface NINJ1 protein(4-8), which contains two transmembrane regions, has an essential role in the induction of PMR. A forward-genetic screen of randomly mutagenized mice linked NINJ1 to PMR. Ninj1(-/-) macrophages exhibited impaired PMR in response to diverse inducers of pyroptotic, necrotic and apoptotic cell death, and were unable to release numerous intracellular proteins including HMGB1 (a known DAMP) and LDH (a standard measure of PMR). Ninj1(-/-) macrophages died, but with a distinctive and persistent ballooned morphology, attributable to defective disintegration of bubble-like herniations. Ninj1(-/-) mice were more susceptible than wild-type mice to infection with Citrobacter rodentium, which suggests a role for PMR in anti-bacterial host defence. Mechanistically, NINJ1 used an evolutionarily conserved extracellular domain for oligomerization and subsequent PMR. The discovery of NINJ1 as a mediator of PMR overturns the long-held idea that cell death-related PMR is a passive event.

Automated summary

Plasma membrane rupture is a cataclysmic event in lytic cell death, releasing damage-associated molecular patterns to propagate the inflammatory response. NINJ1 protein plays an essential role in inducing PMR, with Ninj1-deficient macrophages showing impaired release of intracellular proteins and distinctive ballooned morphology. This study overturns the long-held belief that cell death-related PMR is a passive event, highlighting the active role of NINJ1 in mediating PMR.