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Article ; Online: Measuring Circadian Neutrophil Infiltration in Tissues by Paired Whole-Mount Tissue Clearing and Flow Cytometry.

Vicanolo, Tommaso / Hidalgo, Andres / Adrover, Jose M

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2482, Page(s) 265–284

Abstract: Neutrophils infiltrate most tissues in the organism in the steady state, often following circadian patterns. Neutrophil infiltration is also key to immune defense under inflammatory conditions. In all cases, accurate measurements of the absolute number ... ...

Abstract	Neutrophils infiltrate most tissues in the organism in the steady state, often following circadian patterns. Neutrophil infiltration is also key to immune defense under inflammatory conditions. In all cases, accurate measurements of the absolute number of infiltrated cells and of their localization are important to understand steady-state or inflammatory migration patterns and kinetics. Here we present a method to obtain accurate information on both neutrophil number and distribution that can be successfully applied to circadian studies of neutrophil (or any other cell of interest) migration in vivo. Moreover, this method can be also used to obtain information on activation states or effector functions, for example, by measurement of neutrophil extracellular trap formation in tissues.
MeSH term(s)	Extracellular Traps ; Flow Cytometry ; Neutrophil Infiltration ; Neutrophils
Language	English
Publishing date	2022-05-19
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2249-0_18
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Article ; Online: In Vivo Imaging of Circadian NET Formation During Lung Injury by Four-Dimensional Intravital Microscopy.

Aroca-Crevillén, Alejandra / Hidalgo, Andres / Adrover, Jose M

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2482, Page(s) 285–300

Abstract: Neutrophil extracellular traps (NETs) are toxic extracellular structures deployed by neutrophils in response to pathogens and sterile danger signals. NETs are circadian in nature as mouse and human neutrophils preferentially deploy them at night or early ...

Abstract	Neutrophil extracellular traps (NETs) are toxic extracellular structures deployed by neutrophils in response to pathogens and sterile danger signals. NETs are circadian in nature as mouse and human neutrophils preferentially deploy them at night or early morning. Traditionally, NETs have been quantified using a plethora of methods including immunofluorescence and ELISA-based assays; however few options are available to visualize them in vivo. Here we describe a method to directly visualize and quantify NET formation and release in the microvasculature of the lung using intravital imaging in a model of acute lung injury. The method allows four-dimensional capture and quantification of NET formation dynamics over time and should be a useful resource for those interested in visualizing neutrophil responses in vivo.
MeSH term(s)	Acute Lung Injury/diagnostic imaging ; Animals ; Extracellular Traps ; Intravital Microscopy ; Lung ; Mice ; Neutrophils/physiology
Language	English
Publishing date	2022-05-19
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2249-0_19
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Article ; Online: NETworking with cancer: The bidirectional interplay between cancer and neutrophil extracellular traps.

Adrover, Jose M / McDowell, Sheri A C / He, Xue-Yan / Quail, Daniela F / Egeblad, Mikala

Cancer cell

2023 Volume 41, Issue 3, Page(s) 505–526

Abstract: Neutrophils are major effectors and regulators of the immune system. They play critical roles not only in the eradication of pathogens but also in cancer initiation and progression. Conversely, the presence of cancer affects neutrophil activity, ... ...

Abstract	Neutrophils are major effectors and regulators of the immune system. They play critical roles not only in the eradication of pathogens but also in cancer initiation and progression. Conversely, the presence of cancer affects neutrophil activity, maturation, and lifespan. By promoting or repressing key neutrophil functions, cancer cells co-opt neutrophil biology to their advantage. This co-opting includes hijacking one of neutrophils' most striking pathogen defense mechanisms: the formation of neutrophil extracellular traps (NETs). NETs are web-like filamentous extracellular structures of DNA, histones, and cytotoxic granule-derived proteins. Here, we discuss the bidirectional interplay by which cancer stimulates NET formation, and NETs in turn support disease progression. We review how vascular dysfunction and thrombosis caused by neutrophils and NETs underlie an elevated risk of death from cardiovascular events in cancer patients. Finally, we propose therapeutic strategies that may be effective in targeting NETs in the clinical setting.
MeSH term(s)	Humans ; Extracellular Traps/metabolism ; Neutrophils ; Histones/metabolism ; Thrombosis/etiology ; Thrombosis/metabolism ; DNA/metabolism ; Neoplasms/metabolism
Chemical Substances	Histones ; DNA (9007-49-2)
Language	English
Publishing date	2023-02-23
Publishing country	United States
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2078448-X
ISSN	1878-3686 ; 1535-6108
ISSN (online)	1878-3686
ISSN	1535-6108
DOI	10.1016/j.ccell.2023.02.001
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Article ; Online: Circadian Features of Neutrophil Biology.

Aroca-Crevillén, Alejandra / Adrover, José M / Hidalgo, Andrés

Frontiers in immunology

2020 Volume 11, Page(s) 576

Abstract: Rhythms in immunity manifest in multiple ways, but perhaps most prominently by the recurrent onset of inflammation at specific times of day. These patterns are of importance to understand human disease and are caused, in many instances, by the action of ... ...

Abstract	Rhythms in immunity manifest in multiple ways, but perhaps most prominently by the recurrent onset of inflammation at specific times of day. These patterns are of importance to understand human disease and are caused, in many instances, by the action of neutrophils, a myeloid leukocyte with striking circadian features. The neutrophil's short life, marked diurnal variations in number, and changes in phenotype while in the circulation, help explain the temporal features of inflammatory disease but also uncover core features of neutrophil physiology. Here, we summarize well-established concepts and introduce recent discoveries in the biology of these cells as they relate to circadian rhythms. We highlight that although the circadian features of neutrophils are better known and relevant to understand disease, they may also influence important aspects of organ function even in the steady-state. Finally, we discuss the possibility of targeting these temporal features of neutrophils for therapeutic benefit.
MeSH term(s)	Animals ; Circadian Rhythm/immunology ; Humans ; Neutrophils/physiology
Language	English
Publishing date	2020-04-03
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2020.00576
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Article ; Online: Neutrophils in Homeostasis, Immunity, and Cancer.

Nicolás-Ávila, José Ángel / Adrover, José M / Hidalgo, Andrés

Immunity

2017 Volume 46, Issue 1, Page(s) 15–28

Abstract: Neutrophils were among the first leukocytes described and visualized by early immunologists. Prominent effector functions during infection and sterile inflammation classically placed them low in the immune tree as rapid, mindless aggressors with poor ... ...

Abstract	Neutrophils were among the first leukocytes described and visualized by early immunologists. Prominent effector functions during infection and sterile inflammation classically placed them low in the immune tree as rapid, mindless aggressors with poor regulatory functions. This view is currently under reassessment as we uncover new aspects of their life cycle and identify transcriptional and phenotypic diversity that endows them with regulatory properties that extend beyond their lifetime in the circulation. These properties are revealing unanticipated roles for neutrophils in supporting homeostasis, as well as complex disease states such as cancer. We focus this review on these emerging functions in order to define the true roles of neutrophils in homeostasis, immunity, and disease.
MeSH term(s)	Animals ; Homeostasis/immunology ; Humans ; Immunity, Innate/immunology ; Neoplasms/immunology ; Neutrophils/immunology
Language	English
Publishing date	2017-01-17
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	1217235-2
ISSN	1097-4180 ; 1074-7613
ISSN (online)	1097-4180
ISSN	1074-7613
DOI	10.1016/j.immuni.2016.12.012
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Zs.A 4227: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 69: Show issues

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Article ; Online: Activated platelets jam up the plaque.

Adrover, José M / Hidalgo, Andrés

Circulation research

2015 Volume 116, Issue 4, Page(s) 557–559

MeSH term(s)	Animals ; Aorta/metabolism ; Aortic Diseases/etiology ; Atherosclerosis/etiology ; Blood Platelets/metabolism ; Carotid Artery Diseases/etiology ; Cell Adhesion Molecules/deficiency ; Female ; Humans ; Hyperlipidemias/complications ; Male ; Platelet Aggregation ; Receptors, Cell Surface/deficiency
Chemical Substances	Cell Adhesion Molecules ; F11r protein, mouse ; Receptors, Cell Surface
Language	English
Publishing date	2015-02-13
Publishing country	United States
Document type	Comment ; Editorial ; Research Support, Non-U.S. Gov't
ZDB-ID	80100-8
ISSN	1524-4571 ; 0009-7330 ; 0931-6876
ISSN (online)	1524-4571
ISSN	0009-7330 ; 0931-6876
DOI	10.1161/CIRCRESAHA.115.305823
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Article ; Online: Aging: A Temporal Dimension for Neutrophils.

Adrover, José M / Nicolás-Ávila, José A / Hidalgo, Andrés

Trends in immunology

2016 Volume 37, Issue 5, Page(s) 334–345

Abstract: Neutrophils are first-responders, providing early protection against invading pathogens. Recent findings have revealed a temporal dimension to neutrophil function, associated with the clearance cycles for aging neutrophils, and also with a program that ... ...

Abstract	Neutrophils are first-responders, providing early protection against invading pathogens. Recent findings have revealed a temporal dimension to neutrophil function, associated with the clearance cycles for aging neutrophils, and also with a program that endows circulating neutrophils with distinct phenotypic and functional properties at different times of the day, before they are cleared from blood. We review here the process of neutrophil aging and its impact on homeostasis and inflammation. We outline the features of aged neutrophils, examine proposed mechanisms that drive aging, and discuss how these processes may contribute to tissue homeostasis and pathology. In this context we propose that neutrophil aging may optimize host defense by allowing neutrophils to anticipate infections while avoiding permanent activation and subsequent damage.
MeSH term(s)	Aging/immunology ; Animals ; Homeostasis ; Humans ; Immunity, Innate ; Inflammation/immunology ; Neutrophils/immunology
Language	English
Publishing date	2016-05
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	2036831-8
ISSN	1471-4981 ; 1471-4906
ISSN (online)	1471-4981
ISSN	1471-4906
DOI	10.1016/j.it.2016.03.005
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Zs.A 1601: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Chronic stress increases metastasis via neutrophil-mediated changes to the microenvironment.

Cancer cell

2024 Volume 42, Issue 3, Page(s) 474–486.e12

Abstract: Chronic stress is associated with increased risk of metastasis and poor survival in cancer patients, yet the reasons are unclear. We show that chronic stress increases lung metastasis from disseminated cancer cells 2- to 4-fold in mice. Chronic stress ... ...

Abstract	Chronic stress is associated with increased risk of metastasis and poor survival in cancer patients, yet the reasons are unclear. We show that chronic stress increases lung metastasis from disseminated cancer cells 2- to 4-fold in mice. Chronic stress significantly alters the lung microenvironment, with fibronectin accumulation, reduced T cell infiltration, and increased neutrophil infiltration. Depleting neutrophils abolishes stress-induced metastasis. Chronic stress shifts normal circadian rhythm of neutrophils and causes increased neutrophil extracellular trap (NET) formation via glucocorticoid release. In mice with neutrophil-specific glucocorticoid receptor deletion, chronic stress fails to increase NETs and metastasis. Furthermore, digesting NETs with DNase I prevents chronic stress-induced metastasis. Together, our data show that glucocorticoids released during chronic stress cause NET formation and establish a metastasis-promoting microenvironment. Therefore, NETs could be targets for preventing metastatic recurrence in cancer patients, many of whom will experience chronic stress due to their disease.
MeSH term(s)	Humans ; Animals ; Mice ; Neutrophils/pathology ; Extracellular Traps ; Lung Neoplasms/pathology ; Lung/pathology ; Tumor Microenvironment
Language	English
Publishing date	2024-02-22
Publishing country	United States
Document type	Journal Article
ZDB-ID	2078448-X
ISSN	1878-3686 ; 1535-6108
ISSN (online)	1878-3686
ISSN	1535-6108
DOI	10.1016/j.ccell.2024.01.013
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Article ; Online: Longitudinal Intravital Imaging Through Clear Silicone Windows.

Maiorino, Laura / Shevik, Margaret / Adrover, José M / Han, Xiao / Georgas, Elias / Wilkinson, John Erby / Seidner, Harrison / Foerschner, Leonie / Tuveson, David A / Qin, Yi-Xian / Egeblad, Mikala

Journal of visualized experiments : JoVE

2022 , Issue 179

Abstract: Intravital microscopy (IVM) enables visualization of cell movement, division, and death at single-cell resolution. IVM through surgically inserted imaging windows is particularly powerful because it allows longitudinal observation of the same tissue over ...

Abstract	Intravital microscopy (IVM) enables visualization of cell movement, division, and death at single-cell resolution. IVM through surgically inserted imaging windows is particularly powerful because it allows longitudinal observation of the same tissue over days to weeks. Typical imaging windows comprise a glass coverslip in a biocompatible metal frame sutured to the mouse's skin. These windows can interfere with the free movement of the mice, elicit a strong inflammatory response, and fail due to broken glass or torn sutures, any of which may necessitate euthanasia. To address these issues, windows for long-term abdominal organ and mammary gland imaging were developed from a thin film of polydimethylsiloxane (PDMS), an optically clear silicone polymer previously used for cranial imaging windows. These windows can be glued directly to the tissues, reducing the time needed for insertion. PDMS is flexible, contributing to its durability in mice over time-up to 35 days have been tested. Longitudinal imaging is imaging of the same tissue region during separate sessions. A stainless-steel grid was embedded within the windows to localize the same region, allowing the visualization of dynamic processes (like mammary gland involution) at the same locations, days apart. This silicone window also allowed monitoring of single disseminated cancer cells developing into micro-metastases over time. The silicone windows used in this study are simpler to insert than metal-framed glass windows and cause limited inflammation of the imaged tissues. Moreover, embedded grids allow for straightforward tracking of the same tissue region in repeated imaging sessions.
MeSH term(s)	Animals ; Cell Movement ; Diagnostic Imaging ; Intravital Microscopy/methods ; Mice ; Silicones ; Skull
Chemical Substances	Silicones
Language	English
Publishing date	2022-01-05
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/62757
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Article ; Online: Disulfiram inhibits neutrophil extracellular trap formation and protects rodents from acute lung injury and SARS-CoV-2 infection.

Adrover, Jose M / Carrau, Lucia / Daßler-Plenker, Juliane / Bram, Yaron / Chandar, Vasuretha / Houghton, Sean / Redmond, David / Merrill, Joseph R / Shevik, Margaret / tenOever, Benjamin R / Lyons, Scott K / Schwartz, Robert E / Egeblad, Mikala

JCI insight

2022 Volume 7, Issue 5

Abstract: Severe acute lung injury has few treatment options and a high mortality rate. Upon injury, neutrophils infiltrate the lungs and form neutrophil extracellular traps (NETs), damaging the lungs and driving an exacerbated immune response. Unfortunately, no ... ...

Abstract	Severe acute lung injury has few treatment options and a high mortality rate. Upon injury, neutrophils infiltrate the lungs and form neutrophil extracellular traps (NETs), damaging the lungs and driving an exacerbated immune response. Unfortunately, no drug preventing NET formation has completed clinical development. Here, we report that disulfiram - an FDA-approved drug for alcohol use disorder - dramatically reduced NETs, increased survival, improved blood oxygenation, and reduced lung edema in a transfusion-related acute lung injury (TRALI) mouse model. We then tested whether disulfiram could confer protection in the context of SARS-CoV-2 infection, as NETs are elevated in patients with severe COVID-19. In SARS-CoV-2-infected golden hamsters, disulfiram reduced NETs and perivascular fibrosis in the lungs, and it downregulated innate immune and complement/coagulation pathways, suggesting that it could be beneficial for patients with COVID-19. In conclusion, an existing FDA-approved drug can block NET formation and improve disease course in 2 rodent models of lung injury for which treatment options are limited.
MeSH term(s)	Acetaldehyde Dehydrogenase Inhibitors/pharmacology ; Acute Lung Injury/drug therapy ; Acute Lung Injury/etiology ; Animals ; COVID-19/complications ; COVID-19/virology ; Disease Models, Animal ; Disulfiram/pharmacology ; Extracellular Traps/drug effects ; Extracellular Traps/immunology ; Lung/immunology ; Rodentia ; SARS-CoV-2
Chemical Substances	Acetaldehyde Dehydrogenase Inhibitors ; Disulfiram (TR3MLJ1UAI)
Language	English
Publishing date	2022-03-08
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ISSN	2379-3708
ISSN (online)	2379-3708
DOI	10.1172/jci.insight.157342
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