Activated PMN Exosomes: Pathogenic Entities Causing Matrix Destruction and Disease in the Lung

Genschmer, Kristopher R.; Russell, Derek W.; Lal, Charitharth Vivek; Szul, Tomasz; Bratcher, Preston E.; Noerager, Brett D.; Roda, Mojtaba Abdul; Xu, Xin; Rezonzew, Gabriel; Viera, Liliana; Dobosh, Brian; Margaroli, Camilla; Abdalla, Tarek; King, R. Glenn; McNicholas, Carmel M.; Wells, James M.; Dransfield, Mark T.; Tirouvanziam, Rabindra; Gaggar, Amit; Blalock, J. Edwin

doi:10.1016/j.cell.2018.12.002

Cited by 339 publications

(330 citation statements)

References 53 publications

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“…In pathological states, dysregulation of EV cargo can have profound effects in disease by fundamentally altering the microenvironment and cellular phenotypes through disruption of normal homeostatic pathways while augmenting pathological signals. To this end, several studies reported EVs having a functional role in pulmonary diseases, such as acute lung injury, chronic obstructive pulmonary disease, and lung cancer (25)(26)(27)(28)(29)(30)(31). Only recently have studies begun unraveling the role that EVs play in mediating fibroproliferation (32)(33)(34)(35)(36)(37).…”

Section: Introductionmentioning

confidence: 99%

Syndecan-1 promotes lung fibrosis by regulating epithelial reprogramming through extracellular vesicles

Parimon¹,

Yao²,

Habiel³

et al. 2019

JCI Insight

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Section: Introductionmentioning

confidence: 99%

Syndecan-1 promotes lung fibrosis by regulating epithelial reprogramming through extracellular vesicles

Parimon¹,

Yao²,

Habiel³

et al. 2019

JCI Insight

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“…On the other hand, neutrophil EV-associated effector functions are also known to contribute to innate immune pathology. For example, the surface-bound neutrophil elastase of EVs has been shown to cause extracellular matrix destruction and disease in the lungs of COPD patients and could help explain the antifungal activity of afEVs (57).…”

Section: Discussionmentioning

confidence: 99%

Human neutrophils produce antifungal extracellular vesicles againstAspergillus fumigatus

Shopova

Belyaev

Dasari

et al. 2019

Preprint

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AbstractPolymorphonuclear granulocytes (PMNs) are indispensable for controlling life-threatening fungal infections. In addition to various effector mechanisms, PMNs also produce extracellular vesicles (EVs). Their contribution to antifungal defense has remained unexplored. We reveal that the clinically important human pathogenic fungus Aspergillus fumigatus triggers PMNs to release a distinct set of antifungal EVs (afEVs). Proteome analyses indicated that afEVs are enriched in antimicrobial proteins. The cargo and release kinetics of EVs are modulated by the fungal strain confronted. Tracking of afEVs indicated that they associated with fungal cells and even entered fungal hyphae, resulting in alterations in the morphology of the fungal cell wall and dose-dependent antifungal effects. Two human proteins enriched in afEVs, cathepsin G and azurocidin, were heterologously expressed in fungal hyphae, which led to reduced fungal growth relative to a control retinol binding protein 7 producing strain. In conclusion, the production of afEVs by PMNs offers an intriguing, previously overlooked mechanism of antifungal defense against A. fumigatus.ImportanceInvasive fungal infections caused by the mold Aspergillus fumigatus are a growing concern in the clinic due to the increasing use of immunosuppressive therapies and increasing antifungal drug resistance. These infections result in high mortality as treatment and diagnostic options remain limited. In healthy individuals, neutrophilic granulocytes are critical for elimination of A. fumigatus from the host; however, the exact extracellular mechanism of neutrophil-mediated antifungal activity remains unresolved. Here, we present a mode of antifungal defense employed by human neutrophils against A. fumigatus not previously described. We find that extracellular vesicles produced by neutrophils in response to A. fumigatus infection are able to associate with the fungus, limit growth, and elicit cell damage by delivering antifungal cargo. In the end, antifungal extracellular vesicle biology provides a significant step forward in our understanding of A. fumigatus host pathogenesis and opens up novel diagnostic and therapeutic possibilities.

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“…Our study shows for the first time that the plasma membrane undergoes dramatic changes during NETosis, including shedding of microvesicles and regulated permeabilization. The shedding of microvesicles is of particular interest, as they could be important systemic messengers, providing activated neutrophils undergoing NETosis with additional ways of signaling stress (52) or contributing to disease (53). For instance, microvesicles are procoagulant, thus could potentially contribute to the pro-thrombotic activity of NETs (54).…”

Section: Discussionmentioning

confidence: 99%

NETosis proceeds by cytoskeleton and endomembrane disassembly and PAD4-mediated chromatin de-condensation and nuclear envelope rupture

Waterman¹,

Thiam

Wong

et al. 2019

Preprint

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Neutrophil extracellular traps (NETs) are web-like DNA structures decorated with histones and cytotoxic proteins that are released by activated neutrophils to trap and neutralize pathogens during the innate immune response, but also form in and exacerbate sterile inflammation. Peptidylarginine deiminase 4 (PAD4) citrullinates histones and is required for NET formation (NETosis) in mouse neutrophils. While the in vivo impact of NETs is accumulating, the cellular events driving NETosis and the role of PAD4 in these events are unclear. We performed high resolution time-lapse microscopy of mouse and human neutrophils (PMN) and differentiated HL-60 neutrophil-like cells (dHL-60) labelled with fluorescent markers of organelles and stimulated with ionomycin or lipopolysaccharides to induce NETosis. Upon stimulation, cells exhibited rapid disassembly of the actin cytoskeleton, followed by shedding of plasma membrane microvesicles, disassembly and remodeling of the microtubule and vimentin cytoskeletons, ER vesiculation, chromatin de-condensation and nuclear rounding, progressive plasma membrane and nuclear envelope (NE) permeabilization, nuclear lamin meshwork and then NE rupture to release DNA into the cytoplasm, and finally plasma membrane rupture and discharge of extracellular DNA. Inhibition of actin disassembly blocked NET release. Mouse and dHL-60 cells bearing genetic alteration of PAD4 showed that chromatin de-condensation, lamin meshwork and NE rupture and extracellular DNA release required the enzymatic and nuclear localization activities of PAD4. Thus, NETosis proceeds by a step-wise sequence of cellular events culminating in the PAD4-mediated expulsion of DNA. Significance Statement:Neutrophils are white blood cells specialized as the first line of host defense in the immune system. One way they protect organisms is through NETosis, in which they expel their DNA to form a web-like trap that ensnares pathogens and promotes clotting. However, NETs also mediate sterile inflammation, causing damage to the body. We used high-resolution live-cell microscopy to perform the first systematic characterization of the timing of dynamic cellular events leading to NETosis in human and mouse neutrophils and a neutrophil-like cell line. We discovered that NETosis proceeds by a step-wise sequence of cellular events that is conserved across species, and requires the activity of the PAD4 enzyme for DNA to be released from the nucleus and cell membrane.

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Activated PMN Exosomes: Pathogenic Entities Causing Matrix Destruction and Disease in the Lung

Cited by 339 publications

References 53 publications

Syndecan-1 promotes lung fibrosis by regulating epithelial reprogramming through extracellular vesicles

Syndecan-1 promotes lung fibrosis by regulating epithelial reprogramming through extracellular vesicles

Human neutrophils produce antifungal extracellular vesicles againstAspergillus fumigatus

NETosis proceeds by cytoskeleton and endomembrane disassembly and PAD4-mediated chromatin de-condensation and nuclear envelope rupture

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