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Cited by 857Cell death mechanisms and regulationInflammasome and immune disordersImmune Response and Inflammation

NINJ1 mediates plasma membrane rupture during lytic cell death

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Kayagaki, Nobuhiko, Kornfeld, Opher S., Lee, Bettina L., Stowe, Irma B., O’Rourke, Karen, Li, Qingling, Sandoval, Wendy, Yan, Donghong +19 more·2021·Nature
DOI 10.1038/s41586-021-03218-7

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Abstract

Abstract Plasma membrane rupture (PMR) is the final cataclysmic event in lytic cell death. PMR releases intracellular molecules termed damage-associated molecular patterns (DAMPs) that propagate the inflammatory response. The underlying mechanism for PMR, however, is unknown. Here we show that the ill-characterized nerve injury-induced protein 1 (NINJ1) — a cell surface protein with two transmembrane regions — plays 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 failed to release numerous intracellular proteins including High Mobility Group Box 1 (HMGB1, a known DAMP) and Lactate Dehydrogenase (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 Citrobacter rodentium, suggesting a role for PMR in anti-bacterial host defense. Mechanistically, NINJ1 utilized an evolutionarily conserved extracellular α-helical domain for oligomerization and subsequent PMR. The discovery of NINJ1 as a mediator of PMR overturns the long-held dogma that cell death-related PMR is a passive event. Pyroptosis is a potent inflammatory mode of lytic cell death triggered by diverse infectious and sterile insults1-3. It is driven by the pore-forming fragment of gasdermin D (GSDMD)4-7 and releases two exemplar proteins: interleukin-1β (IL-1β), a pro-inflammatory cytokine, and LDH, a standard marker of PMR and lytic cell death. An early landmark study8 predicted two sequential steps for pyroptosis: (1) initial formation of a small plasma membrane pore causing IL-1β release and non-selective ionic fluxes, and (2) subsequent PMR attributable to oncotic cell swelling. PMR releases LDH (140 kDa) and large DAMPs. While the predicted size of gasdermin pores (~18 nm inner diameter9) is large enough to release IL-1β (17 kDa, ~4.5 nm diameter), the underlying mechanism for subsequent PMR has been considered a passive osmotic lysis event.

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  • Cell membrane rupture upon pyroptosis leads to the release of DAMPs such as HMGB1 as well as ASC specks into the extracellular space.

    “Cell membrane rupture upon pyroptosis leads to the release of DAMPs such as HMGB1 as well as ASC specks into the extracellular space [69, 70].”

    From Microglial function, INPP5D/SHIP1 signaling, and NLRP3 inflammasome activation: implications for Alzheimer’s disease (2023) · cites this paper as [69]

    The Abstract and “NINJ1 is essential for pyroptosis-related PMR” section report that plasma membrane rupture releases DAMPs, including HMGB1, from pyroptotic cells. Our copy does not establish that rupture also releases ASC specks; a search for “ASC specks” found no supporting passage. Page numbers were not available in the indexed copy. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • Pyroptosis may occur downstream of GSDMD pore formation and is mediated by Ninjurin1, a cell surface protein that induces plasma membrane rupture upon its oligomerization.

    “Pyroptosis may occur downstream of GSDMD pore formation and is mediated by Ninjurin1 (NINJ1), a cell surface protein that induces plasma membrane rupture upon its oligomerization [68, 69].”

    From Microglial function, INPP5D/SHIP1 signaling, and NLRP3 inflammasome activation: implications for Alzheimer’s disease (2023) · cites this paper as [69]

    The Abstract and ‘NINJ1 oligomerizes for induction of PMR’ section support that NINJ1 is a cell-surface protein whose oligomerization induces plasma membrane rupture. But ‘NINJ1 is essential for pyroptosis-related PMR’ distinguishes that rupture from GSDMD-driven cell death: NINJ1 is required for the former, not pyroptosis as a whole. Our copy provides no page numbers for these passages. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

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Cited by

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Authors
Kayagaki, N., Kornfeld, O. S., Lee, B. L., Stowe, I. B., O’Rourke, K., Li, Q., Sandoval, W., Yan, D., Kang, J., Xu, M., Zhang, J., Lee, W. P., McKenzie, B. S., Ulas, G., Payandeh, J., Roose-Girma, M., Modrusan, Z., Reja, R., Sagolla, M., Webster, J. D., Cho, V., Andrews, T. D., Morris, L. X., Miosge, L. A., Goodnow, C. C., Bertram, E. M., & Dixit, V. M.
Affiliations
  • Genentech (US)
  • Australian National University (AU)
  • Garvan Institute of Medical Research (AU)
Corresponding author
  • Kayagaki, Nobuhiko
  • Dixit, Vishva M.
ORCID
Venue
Nature
Publisher
Springer Science and Business Media LLC
Published
2021-01-20
Volume
591
Issue
7848
Pages
131-136
ISSN
  • 0028-0836 (ISSN-L)
  • 1476-4687
DOI
10.1038/s41586-021-03218-7
OpenAlex
W3115667040
Cited by
857
References
6
Citations by year
2026: 199 · 2025: 182 · 2024: 150 · 2023: 135 · 2022: 138 · 2021: 51 · 2020: 1
FWCI
38.82
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature (publishedVersion)
  • Research Square (acceptedVersion, cc-by)
  • PubMed (publishedVersion)
  • ANU Open Research (Australian National University) (submittedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
Funders
Genentech
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Inflammasome and immune disorders
  • Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
Keywords
NINJ1, plasma membrane rupture, lytic cell death, pyroptosis, necrosis, apoptosis, damage-associated molecular patterns, HMGB1, lactate dehydrogenase release, macrophages, Citrobacter rodentium infection, protein oligomerization, extracellular domain, forward genetic screen, inflammation
MeSH terms
Pyroptosis, Animals, Cell Membrane, Female, Humans, Macrophages, Male, Mutation, Necrosis, Nerve Growth Factors, Cell Adhesion Molecules, Neuronal, Cell Death, Apoptosis, Mice, Protein Multimerization

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