Gene silencing of endothelial von Willebrand factor reduces the susceptibility of human endothelial cells to SARS-CoV-2 infection.

Autor: Furini G; Unit of Translational Critical Care Medicine, Laboratory of Basic and Applied Medical Sciences, Interdisciplinary Research Center 'Health Science', Scuola Superiore Sant'Anna, Pisa, Italy.; UOSVD Anesthesia and Resuscitation, Fondazione Toscana 'G. Monasterio', Pisa, Italy., De Carli A; Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, Retrovirus Center, University of Pisa, Italy., Fonnesu R; Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, Retrovirus Center, University of Pisa, Italy., Spezia PG; Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, Retrovirus Center, University of Pisa, Italy., Scebba F; BioMedicine Laboratory, Interdisciplinary Research Center 'Health Science', Scuola Superiore Sant'Anna, Pisa, Italy., Pistello M; Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, Retrovirus Center, University of Pisa, Italy.; Virology Unit, Pisa University Hospital, Italy., Lai M; Virology Section, Department of Translational Research and New Technologies in Medicine and Surgery, Retrovirus Center, University of Pisa, Italy.; CISUP - Centre for Instrumentation Sharing - University of Pisa, Italy., Lionetti V; Unit of Translational Critical Care Medicine, Laboratory of Basic and Applied Medical Sciences, Interdisciplinary Research Center 'Health Science', Scuola Superiore Sant'Anna, Pisa, Italy.; UOSVD Anesthesia and Resuscitation, Fondazione Toscana 'G. Monasterio', Pisa, Italy.; BioMedicine Laboratory, Interdisciplinary Research Center 'Health Science', Scuola Superiore Sant'Anna, Pisa, Italy.
Jazyk: angličtina
Zdroj: The FEBS journal [FEBS J] 2023 Sep; Vol. 290 (17), pp. 4300-4315. Date of Electronic Publication: 2023 May 10.
DOI: 10.1111/febs.16808
Abstrakt: Mechanisms underlying vascular endothelial susceptibility to infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are not fully understood. Emerging evidence indicates that patients lacking von Willebrand factor (vWF), an endothelial hallmark, are less severely affected by SARS-CoV-2 infection, yet the precise role of endothelial vWF in modulating coronavirus entry into endothelial cells is unknown. In the present study, we demonstrated that effective gene silencing by short interfering RNA (siRNA) for vWF expression in resting human umbilical vein endothelial cells (HUVECs) significantly reduced by 56% the cellular levels of SARS-CoV-2 genomic RNA. Similar reduction in intracellular SARS-CoV-2 genomic RNA levels was observed in non-activated HUVECs treated with siRNA targeting angiotensin-converting enzyme 2 (ACE2), the cellular gateway to coronavirus. By integrating quantitative information from real-time PCR and high-resolution confocal imaging, we demonstrated that ACE2 gene expression and its plasma membrane localization in HUVECs were both markedly reduced after treatment with siRNA anti-vWF or anti-ACE2. Conversely, siRNA anti-ACE2 did not reduce endothelial vWF gene expression and protein levels. Finally, SARS-CoV-2 infection of viable HUVECs was enhanced by overexpression of vWF, which increased ACE2 levels. Of note, we found a similar increase in interferon-β mRNA levels following transfection with untargeted, anti-vWF or anti-ACE2 siRNA and pcDNA3.1-WT-VWF. We envision that siRNA targeting endothelial vWF will protect against productive endothelial infection by SARS-CoV-2 through downregulation of ACE2 expression and might serve as a novel tool to induce disease resistance by modulating the regulatory role of vWF on ACE2 expression.
(© 2023 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)
Databáze: MEDLINE
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