Flavivirus NS1 Triggers Tissue-Specific Disassembly of Intercellular Junctions Leading to Barrier Dysfunction and Vascular Leak in a GSK-3β-Dependent Manner

Puerta-Guardo, Henry; Biering, Scott B.; Sousa, Francielle Tramontini Gomes de; Shu, Jeffrey; Glasner, Dustin R.; Li, Jeffrey; Blanc, Sophie F.; Beatty, P. Robert; Harris, Eva

doi:10.3390/pathogens11060615

Cited by 18 publications

(22 citation statements)

References 98 publications

(169 reference statements)

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“…We and others have described a phenomenon by which viral proteins, such as the flavivirus non-structural protein 1 (NS1), interact with endothelial cells to trigger signaling cascades that mediate disruption of cellular structures required for endothelial barrier integrity, including the endothelial glycocalyx layer (EGL) and intercellular junctional complexes 29 – 34 . Here, we investigated whether SARS-CoV-2 S contributes to endothelial and epithelial barrier dysfunction in vitro and vascular leak in vivo.…”

Section: Introductionmentioning

confidence: 99%

SARS-CoV-2 Spike triggers barrier dysfunction and vascular leak via integrins and TGF-β signaling

et al. 2022

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Severe COVID-19 is associated with epithelial and endothelial barrier dysfunction within the lung as well as in distal organs. While it is appreciated that an exaggerated inflammatory response is associated with barrier dysfunction, the triggers of vascular leak are unclear. Here, we report that cell-intrinsic interactions between the Spike (S) glycoprotein of SARS-CoV-2 and epithelial/endothelial cells are sufficient to induce barrier dysfunction in vitro and vascular leak in vivo, independently of viral replication and the ACE2 receptor. We identify an S-triggered transcriptional response associated with extracellular matrix reorganization and TGF-β signaling. Using genetic knockouts and specific inhibitors, we demonstrate that glycosaminoglycans, integrins, and the TGF-β signaling axis are required for S-mediated barrier dysfunction. Notably, we show that SARS-CoV-2 infection caused leak in vivo, which was reduced by inhibiting integrins. Our findings offer mechanistic insight into SARS-CoV-2-triggered vascular leak, providing a starting point for development of therapies targeting COVID-19.

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Section: Introductionmentioning

confidence: 99%

SARS-CoV-2 Spike triggers barrier dysfunction and vascular leak via integrins and TGF-β signaling

et al. 2022

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“…NS1, however, exists as an intracellular monomer, a membrane-bound dimer, and an extracellular hexamer ( Rastogi et al., 2016 ). During infection, secreted NS1 disrupts endothelial junctions, resulting in tissue permeabilization and vascular leakage ( Puerta-Guardo et al., 2019 ; Puerta-Guardo et al., 2022 ), so neutralizing NS1 may be an important protective strategy. Indeed, passive transfer studies using serum from mice vaccinated with NS1 protected against a lethal ZIKV challenge ( Bailey et al., 2019 ).…”

Section: Discussionmentioning

confidence: 99%

Replication in the presence of dengue convalescent serum impacts Zika virus neutralization sensitivity and fitness

Marano

Weger-Lucarelli

2023

Front. Cell. Infect. Microbiol.

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IntroductionFlaviviruses like dengue virus (DENV) and Zika virus (ZIKV) are mosquito-borne viruses that cause febrile, hemorrhagic, and neurological diseases in humans, resulting in 400 million infections annually. Due to their co-circulation in many parts of the world, flaviviruses must replicate in the presence of pre-existing adaptive immune responses targeted at serologically closely related pathogens, which can provide protection or enhance disease. However, the impact of pre-existing cross-reactive immunity as a driver of flavivirus evolution, and subsequently the implications on the emergence of immune escape variants, is poorly understood. Therefore, we investigated how replication in the presence of convalescent dengue serum drives ZIKV evolution.MethodsWe used an in vitro directed evolution system, passaging ZIKV in the presence of serum from humans previously infected with DENV (anti-DENV) or serum from DENV-naïve patients (control serum). Following five passages in the presence of serum, we performed next-generation sequencing to identify mutations that arose during passaging. We studied two non-synonymous mutations found in the anti-DENV passaged population (E-V355I and NS1-T139A) by generating individual ZIKV mutants and assessing fitness in mammalian cells and live mosquitoes, as well as their sensitivity to antibody neutralization.Results and discussionBoth viruses had increased fitness in Vero cells with and without the addition of anti-DENV serum and in human lung epithelial and monocyte cells. In Aedes aegypti mosquitoes—using blood meals with and without anti-DENV serum—the mutant viruses had significantly reduced fitness compared to wild-type ZIKV. These results align with the trade-off hypothesis of constrained mosquito-borne virus evolution. Notably, only the NS1-T139A mutation escaped neutralization, while E-V335I demonstrated enhanced neutralization sensitivity to neutralization by anti-DENV serum, indicating that neutralization escape is not necessary for viruses passaged under cross-reactive immune pressures. Future studies are needed to assess cross-reactive immune selection in humans and relevant animal models or with different flaviviruses.

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“…NS1, however, exists as an intracellular monomer, a membrane-bound dimer, and an extracellular hexamer [78]. During infection, secreted NS1 disrupts endothelial junctions, resulting in tissue permeabilization and vascular leakage [79, 80], so neutralizing NS1 may be an important protective strategy. Indeed, passive transfer studies using serum from mice vaccinated with NS1 protected against a lethal ZIKV challenge [81].…”

Section: Discussionmentioning

confidence: 99%

Replication in the presence of dengue convalescent serum impacts Zika virus neutralization sensitivity and fitness

Marano

Weger-Lucarelli

2022

Preprint

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Flaviviruses like dengue virus (DENV) and Zika virus (ZIKV) are mosquito-borne viruses that cause febrile, hemorrhagic, and neurological diseases in humans, resulting in 400 million infections annually. Due to their co-circulation in many parts of the world, flaviviruses must replicate in the presence of pre-existing adaptive immune responses targeted at serologically closely related pathogens, which can provide protection or enhance disease. However, the impact of pre-existing cross-reactive immunity as a driver of flavivirus evolution, and subsequently the implications on the emergence of immune escape variants, is poorly understood. Therefore, we investigated how replication in the presence of convalescent dengue serum drives ZIKV evolution. We used anin vitrodirected evolution system, passaging ZIKV in the presence of serum from humans previously infected with DENV (anti-DENV) or serum from DENV-na&iumlve patients (control serum). Following five passages in the presence of serum, we performed next-generation sequencing to identify mutations that arose during passaging. We studied two non-synonymous mutations found in the anti-DENV passaged population (E-V355I and NS1-T139A) by generating individual ZIKV mutants and assessing fitness in mammalian cells and live mosquitoes, as well as their sensitivity to antibody neutralization. Both viruses had increased fitness in Vero cells with and without the addition of anti-DENV serum and in human lung epithelial and monocyte cells. InAedes aegyptimosquitoes — using blood meals with and without anti-DENV serum — the mutant viruses had significantly reduced fitness compared to wild-type ZIKV. These results align with the trade-off hypothesis of constrained mosquito-borne virus evolution. Notably, only the NS1-T139A mutation escaped neutralization, while E-V335I demonstrated enhanced neutralization sensitivity to neutralization by anti-DENV serum, indicating that neutralization escape is not necessary for viruses passaged under cross-reactive immune pressures. Future studies are needed to assess cross-reactive immune selection in humans and relevant animal models or with different flaviviruses.

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Flavivirus NS1 Triggers Tissue-Specific Disassembly of Intercellular Junctions Leading to Barrier Dysfunction and Vascular Leak in a GSK-3β-Dependent Manner

Cited by 18 publications

References 98 publications

SARS-CoV-2 Spike triggers barrier dysfunction and vascular leak via integrins and TGF-β signaling

SARS-CoV-2 Spike triggers barrier dysfunction and vascular leak via integrins and TGF-β signaling

Replication in the presence of dengue convalescent serum impacts Zika virus neutralization sensitivity and fitness

Replication in the presence of dengue convalescent serum impacts Zika virus neutralization sensitivity and fitness

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