Dataset Information

Nexinhib20 Inhibits Neutrophil Adhesion and β₂ Integrin Activation by Antagonizing Rac-1-Guanosine 5'-Triphosphate Interaction.

ABSTRACT: Neutrophils are critical for mediating inflammatory responses. Inhibiting neutrophil recruitment is an attractive approach for preventing inflammatory injuries, including myocardial ischemia-reperfusion (I/R) injury, which exacerbates cardiomyocyte death after primary percutaneous coronary intervention in acute myocardial infarction. In this study, we found out that a neutrophil exocytosis inhibitor Nexinhib20 inhibits not only exocytosis but also neutrophil adhesion by limiting β₂ integrin activation. Using a microfluidic chamber, we found that Nexinhib20 inhibited IL-8-induced β₂ integrin-dependent human neutrophil adhesion under flow. Using a dynamic flow cytometry assay, we discovered that Nexinhib20 suppresses intracellular calcium flux and β₂ integrin activation after IL-8 stimulation. Western blots of Ras-related C3 botulinum toxin substrate 1 (Rac-1)-GTP pull-down assays confirmed that Nexinhib20 inhibited Rac-1 activation in leukocytes. An in vitro competition assay showed that Nexinhib20 antagonized the binding of Rac-1 and GTP. Using a mouse model of myocardial I/R injury, Nexinhib20 administration after ischemia and before reperfusion significantly decreased neutrophil recruitment and infarct size. Our results highlight the translational potential of Nexinhib20 as a dual-functional neutrophil inhibitory drug to prevent myocardial I/R injury.

SUBMITTER: Liu W

PROVIDER: S-EPMC9529951 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature

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Nexinhib20 Inhibits Neutrophil Adhesion and β<sub>2</sub> Integrin Activation by Antagonizing Rac-1-Guanosine 5'-Triphosphate Interaction.

Liu Wei W Cronin Chunxia G CG Cao Ziming Z Wang Chengliang C Ruan Jianbin J Pulikkot Sunitha S Hall Alexxus A Sun Hao H Groisman Alex A Chen Yunfeng Y Vella Anthony T AT Hu Liang L Liang Bruce T BT Fan Zhichao Z

Journal of immunology (Baltimore, Md. : 1950) 20220907 8

Neutrophils are critical for mediating inflammatory responses. Inhibiting neutrophil recruitment is an attractive approach for preventing inflammatory injuries, including myocardial ischemia-reperfusion (I/R) injury, which exacerbates cardiomyocyte death after primary percutaneous coronary intervention in acute myocardial infarction. In this study, we found out that a neutrophil exocytosis inhibitor Nexinhib20 inhibits not only exocytosis but also neutrophil adhesion by limiting β<sub>2</sub> in ...[more]

PMID: 36165184

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Project description:Neutrophil extracellular traps (NETs), originally recognized as a host defense mechanism, were reported to promote thrombosis and metastatic dissemination of cancer cells. Here we tested the role of integrins α5β1 and ανβ3 in the adhesion of cancer cells to NETs. Neutrophil-like cells stimulated with calcium ionophore (A23187) were used as a stable source of cell-free NETs-enriched suspensions. Using NETs as an adhesion substrate, two human K562 cell lines, differentially expressing α5β1 and ανβ3 integrins, were subjected to adhesion assays in the presence or absence of DNAse 1, blocking antibodies against α5β1 or ανβ3, alone or in combination with DNAse 1, and Proteinase K. As expected DNAse 1 treatment strongly inhibited adhesion of both cell lines to NETs. An equivalent significant reduction of cell adhesion to NETs was obtained after treatment of cells with blocking antibodies against α5β1 or ανβ3 indicating that both integrins were able to mediate cell adhesion to NETs. Furthermore, the combination of DNAse 1 and anti-integrin antibody treatment almost completely blocked cell adhesion. Western blot analysis and immunoprecipitation experiments showed a dose-dependent increase of fibronectin levels in samples from stimulated neutrophil-like cells and a direct or indirect interaction of fibronectin with histone H3. Finally, co-immunolocalization studies with confocal microscopy showed that fibronectin and citrullinated histone H3 co-localize inside the web-structure of NETs. In conclusion, our study showed that α5β1 and ανβ3 integrins mediate cell adhesion to NETs by binding to their common substrate fibronectin. Therefore, in addition to mechanical trapping and aspecific adsorption of different cell types driven by DNA/histone complexes, NETs may provide specific binding sites for integrin-mediated cell adhesion of neutrophils, platelets, endothelial and cancer cells thus promoting intimate interactions among these cells.

Dataset Information

Nexinhib20 Inhibits Neutrophil Adhesion and β₂ Integrin Activation by Antagonizing Rac-1-Guanosine 5'-Triphosphate Interaction.

Publications

Nexinhib20 Inhibits Neutrophil Adhesion and β<sub>2</sub> Integrin Activation by Antagonizing Rac-1-Guanosine 5'-Triphosphate Interaction.

Similar Datasets

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Dataset Information

Nexinhib20 Inhibits Neutrophil Adhesion and β2 Integrin Activation by Antagonizing Rac-1-Guanosine 5'-Triphosphate Interaction.

Publications

Nexinhib20 Inhibits Neutrophil Adhesion and β&lt;sub&gt;2&lt;/sub&gt; Integrin Activation by Antagonizing Rac-1-Guanosine 5'-Triphosphate Interaction.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Nexinhib20 Inhibits Neutrophil Adhesion and β₂ Integrin Activation by Antagonizing Rac-1-Guanosine 5'-Triphosphate Interaction.

Nexinhib20 Inhibits Neutrophil Adhesion and β<sub>2</sub> Integrin Activation by Antagonizing Rac-1-Guanosine 5'-Triphosphate Interaction.