Visualization of perforin/gasdermin/complement-formed pores in real cell membranes using atomic force microscopy

Liu, Yuying; Zhang, Tianzhen; Zhou, Yabo; Li, Jiping; Liang, Xiaoyu; Zhou, Nannan; Lv, Jiadi; Xie, Jing; Cheng, Feiran; Fang, Yiliang; Gao, Yunfeng; Wang, Ning; Huang, Bo

doi:10.1038/s41423-018-0165-1

Cited by 39 publications

(43 citation statements)

References 34 publications

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“…Pyroptosis is a newly discovered form of programmed lytic cell death that is induced by inflammatory casapse-1 (CASP1), and is characterized by swelling of the cell, pore formation in unilamellar liposomes, membrane rupture and the release of inflammatory cytokines [14][15][16][17][18]. Gasdermin D (GSDMD) was discovered to be a substrate of CASP1, and cleavage of the N-terminal fragment of GSDMD forms membrane pores and drives pyroptosis [19][20][21]. Inflammasome complexes function as crucial intracellular effectors to initiate innate immunity and thus to defend against infections or tissue damage [22,23].…”

Section: Introductionmentioning

confidence: 99%

NLRP12 collaborates with NLRP3 and NLRC4 to promote pyroptosis inducing ganglion cell death of acute glaucoma

Chen

Deng

Gan

et al. 2020

Mol Neurodegeneration

109

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Background: Acute glaucoma, characterized by a sudden elevation in intraocular pressure (IOP) and retinal ganglion cells (RGCs) death, is a major cause of irreversible blindness worldwide that lacks approved effective therapies, validated treatment targets and clear molecular mechanisms. We sought to explore the potential molecular mechanisms underlying the causal link between high IOP and glaucomatous RGCs death.Methods: A murine retinal ischemia/ reperfusion (RIR) model and an in vitro oxygen and glucose deprivation/ reoxygenation (OGDR) model were used to investigate the pathogenic mechanisms of acute glaucoma.Results: Our findings reveal a novel mechanism of microglia-induced pyroptosis-mediated RGCs death associated with glaucomatous vision loss. Genetic deletion of gasdermin D (GSDMD), the effector of pyroptosis, markedly ameliorated the RGCs death and retinal tissue damage in acute glaucoma. Moreover, GSDMD cleavage of microglial cells was dependent on caspase-8 (CASP8)-hypoxia-inducible factor-1α (HIF-1α) signaling. Mechanistically, the newly identified nucleotide-binding leucine-rich repeat-containing receptor (NLR) family pyrin domain-containing 12 (NLRP12) collaborated with NLR family pyrin domain-containing 3 (NLRP3) and NLR family CARD domain-containing protein 4 (NLRC4) downstream of the CASP8-HIF-1α axis, to elicit pyroptotic processes and interleukin-1β (IL-1β) maturation through caspase-1 activation, facilitating pyroptosis and neuroinflammation in acute glaucoma. Interestingly, processing of IL-1β in turn magnified the CASP8-HIF-1α-NLRP12/ NLRP3/NLRC4-pyroptosis circuit to accelerate inflammatory cascades.Conclusions: These data not only indicate that the collaborative effects of NLRP12, NLRP3 and NLRC4 on pyroptosis are responsible for RGCs death, but also shed novel mechanistic insights into microglial pyroptosis, paving novel therapeutic avenues for the treatment of glaucoma-induced irreversible vision loss through simultaneously targeting of pyroptosis.

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Section: Introductionmentioning

confidence: 99%

NLRP12 collaborates with NLRP3 and NLRC4 to promote pyroptosis inducing ganglion cell death of acute glaucoma

Chen

Deng

Gan

et al. 2020

Mol Neurodegeneration

109

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“…High-resolution atomic force microscopy studies with GSDMD-N showed that monomers can first associate forming arcs or slit structures that later evolve into transmembrane pores with a ring shape. Notably, all these structures are able to perforate the membranes forming holes in the 10-30 nm range [28]. The fact that incomplete rings of GSDMD-N promote the opening of the membrane clearly indicates the direct involvement of lipids in pore opening.…”

Section: Pore Formation By Gsdms In Pyroptosismentioning

confidence: 96%

“…The lipid membrane provides a suitable environment to trigger the conformational changes required for their structural evolution from soluble monomers to membrane-embedded oligomers [27]. In recent years, our understanding of the mechanism of GSDMs pore formation has dramatically increased with the structural characterization of different variants in solution [10] and in membranes [27,28]. The analysis of the crystal structure of full-length GSDMA3 [10] and the cryo-electron microscopy structure of a GSDMA3-N pore isolated from reconstituted liposomes [27] shed light on the mechanisms of membrane insertion and pore assembly.…”

Section: Pore Formation By Gsdms In Pyroptosismentioning

confidence: 99%

Partners in Crime: The Interplay of Proteins and Membranes in Regulated Necrosis

Ros

Pedrera

García‐Sáez

2020

IJMS

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Pyroptosis, necroptosis, and ferroptosis are well-characterized forms of regulated necrosis that have been associated with human diseases. During regulated necrosis, plasma membrane damage facilitates the movement of ions and molecules across the bilayer, which finally leads to cell lysis and release of intracellular content. Therefore, these types of cell death have an inflammatory phenotype. Each type of regulated necrosis is mediated by a defined machinery comprising protein and lipid molecules. Here, we discuss how the interaction and reshaping of these cellular components are essential and distinctive processes during pyroptosis, necroptosis, and ferroptosis. We point out that although the plasma membrane is the common target in regulated necrosis, different mechanisms of permeabilization have emerged depending on the cell death form. Pore formation by gasdermins (GSDMs) is a hallmark of pyroptosis, while mixed lineage kinase domain-like (MLKL) protein facilitates membrane permeabilization in necroptosis, and phospholipid peroxidation leads to membrane damage in ferroptosis. This diverse repertoire of mechanisms leading to membrane permeabilization contributes to define the specific inflammatory and immunological outcome of each type of regulated necrosis. Current efforts are focused on new therapies that target critical protein and lipid molecules on these pathways to fight human pathologies associated with inflammation.

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“…Author Manuscript Published OnlineFirst on November 9, 2020; DOI: 10.1158/0008-5472. CAN-20-2569 The AFM tips (NPG-10) were functionalized with anti-PFR or anti-SLO antibody as described before (21). In brief, NPG-10 tips were coated with the crosslinker of NHS-PEG-maleimide for 1h at room temperature.…”

Section: Downloaded Frommentioning

confidence: 99%

Cell Softness Prevents Cytolytic T-cell Killing of Tumor-Repopulating Cells

Liu

Zhang

et al. 2021

Cancer Research

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Biomechanics is a fundamental feature of a cell. However, the manner by which actomysin tension affects tumor immune evasion remains unclear. Here we show that although cytotoxic T lymphocytes (CTL) can effectively destroy stiff differentiated tumor cells, they fail to kill soft tumor-repopulating cells (TRC). TRC softness prevented membrane pore formation caused by CTL-released perforin. Perforin interacting with nonmuscle myosin heavy-chain 9 transmitted forces to less F-actins in soft TRC, thus generating an inadequate contractile force for perforin pore formation. Stiffening TRC allowed perforin the ability to drill through the membrane, leading to CTL-mediated killing of TRC. Importantly, overcoming mechanical softness in human TRC also enhanced TRC cell death caused by human CTL, potentiating a mechanics-based immunotherapeutic strategy. These findings reveal a mechanics-mediated tumor immune evasion, thus potentially providing an alternative approach for tumor immunotherapy. Significance: Tumor-repopulating cells evade CD8+ cytolytic T-cell killing through a mechanical softness mechanism, underlying the impediment of perforin pore formation at the immune synapse site.

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Visualization of perforin/gasdermin/complement-formed pores in real cell membranes using atomic force microscopy

Cited by 39 publications

References 34 publications

NLRP12 collaborates with NLRP3 and NLRC4 to promote pyroptosis inducing ganglion cell death of acute glaucoma

NLRP12 collaborates with NLRP3 and NLRC4 to promote pyroptosis inducing ganglion cell death of acute glaucoma

Partners in Crime: The Interplay of Proteins and Membranes in Regulated Necrosis

Cell Softness Prevents Cytolytic T-cell Killing of Tumor-Repopulating Cells

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