Zika Virus Infects Trabecular Meshwork and Causes Trabeculitis and Glaucomatous Pathology in Mouse Eyes

ABSTRACT Zika virus (ZIKV) infection during pregnancy leads to devastating fetal outcomes, including neurological (microcephaly) and ocular pathologies such as retinal lesions, optic nerve abnormalities, chorioretinal atrophy, and congenital glaucoma. Only clinical case reports have linked ZIKV infection to causing glaucoma, a major blinding eye disease. In the present study, we have investigated the role of ZIKV in glaucoma pathophysiology using in vitro and in vivo experimental models. We showed that human primary trabecular meshwork (Pr. TM) cells, as well as a human GTM3 cell line, were permissive to ZIKV infection. ZIKV induced the transcription of various genes expressing pattern recognition receptors (TLR2, TLR3, and RIG-I), cytokines/chemokines (TNF-α, IL-1β, CCL5, and CXCL10), interferons (IFN-α2, IFN-β1, and IFN-γ), and interferon-stimulated genes (ISG15 and OAS2) in Pr. TM cells. ZIKV infection in IFNAR1−/− and wild-type (WT) mouse eyes resulted in increased intraocular pressure (IOP) and the development of chorioretinal atrophy. Anterior chamber (AC) inoculation of ZIKV caused infectivity in iridocorneal angle and TM, leading to the death of TM cells in the mouse eyes. Moreover, anterior segment tissue of infected eyes exhibited increased expression of inflammatory mediators and interferons. Furthermore, ZIKV infection in IFNAR1−/− mice resulted in retinal ganglion cell (RGC) death and loss, coinciding with optic nerve infectivity and disruption of anterograde axonal transport. Because of similarity in glaucomatous pathologies in our study and other experimental glaucoma models, ZIKV infection can be used to study infectious triggers of glaucoma, currently an understudied area of investigation. IMPORTANCE Ocular complications due to ZIKV infection remains a major public health concern because of their ability to cause visual impairment or blindness. Most of the previous studies have shown ZIKV-induced ocular pathology in the posterior segment (i.e., retina) of the eye. However, some recent clinical reports from affected countries highlighted the importance of ZIKV in affecting the anterior segment of the eye and causing congenital glaucoma. Because glaucoma is the second leading cause of blindness worldwide, it is imperative to study ZIKV infection in causing glaucoma to identify potential targets for therapeutic intervention. In this study, we discovered that ZIKV permissively infects human TM cells and evokes inflammatory responses causing trabeculitis. Using a mouse model, we demonstrated that ZIKV infection resulted in higher IOP, increased RGC loss, and optic nerve abnormalities, the classical hallmarks of glaucoma. Collectively, our study provides new insights into ocular ZIKV infection resulting in glaucomatous pathology.

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Autophagy in Zika Virus Infection: A Possible Therapeutic Target to Counteract Viral Replication

International Journal of Molecular Sciences ◽

10.3390/ijms20051048 ◽

2019 ◽

Vol 20 (5) ◽

pp. 1048 ◽

Cited By ~ 11

Author(s):

Rossella Gratton ◽

Almerinda Agrelli ◽

Paola Tricarico ◽

Lucas Brandão ◽

Sergio Crovella

Keyword(s):

Public Health ◽

Viral Replication ◽

Zika Virus ◽

Health Concern ◽

Mtor Signaling Pathway ◽

Catabolic Pathway ◽

Public Health Concern ◽

Zikv Infection ◽

Fundamental Process

Zika virus (ZIKV) still constitutes a public health concern, however, no vaccines or therapies are currently approved for treatment. A fundamental process involved in ZIKV infection is autophagy, a cellular catabolic pathway delivering cytoplasmic cargo to the lysosome for degradation—considered as a primordial form of innate immunity against invading microorganisms. ZIKV is thought to inhibit the Akt-mTOR signaling pathway, which causes aberrant activation of autophagy promoting viral replication and propagation. It is therefore appealing to study the role of autophagic molecular effectors during viral infection to identify potential targets for anti-ZIKV therapeutic intervention.

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Zika virus is transmitted in neural progenitor cells via cell-to-cell spread and infection is inhibited by the autophagy inducer trehalose

Journal of Virology ◽

10.1128/jvi.02024-20 ◽

2020 ◽

pp. JVI.02024-20

Author(s):

Alex E Clark ◽

Zhe Zhu ◽

Florian Krach ◽

Jeremy N Rich ◽

Gene W. Yeo ◽

...

Keyword(s):

Viral Replication ◽

Progenitor Cells ◽

Zika Virus ◽

Neural Progenitor Cells ◽

Neutralizing Antibody ◽

Neural Progenitor ◽

Health Concern ◽

Free Virus ◽

Zikv Infection ◽

Infected Cells

Zika virus (ZIKV) is a mosquito-borne human pathogen that causes congenital Zika syndrome and neurological symptoms in some adults. There are currently no approved treatments or vaccines for ZIKV, and exploration of therapies targeting host processes could avoid viral development of drug resistance. The purpose of our study was to determine if the non-toxic and widely used disaccharide trehalose, which showed antiviral activity against Human Cytomegalovirus (HCMV) in our previous work, could restrict ZIKV infection in clinically relevant neural progenitor cells (NPCs). Trehalose is known to induce autophagy, the degradation and recycling of cellular components. Whether autophagy is proviral or antiviral for ZIKV is controversial and depends on cell type and specific conditions used to activate or inhibit autophagy. We show here that trehalose treatment of NPCs infected with recent ZIKV isolates from Panama and Puerto Rico significantly reduces viral replication and spread. In addition, we demonstrate that ZIKV infection in NPCs spreads primarily cell-to-cell as an expanding infectious center, and NPCs are infected via contact with infected cells far more efficiently than by cell-free virus. Importantly, ZIKV was able to spread in NPCs in the presence of neutralizing antibody.Importance Zika virus causes birth defects and can lead to neurological disease in adults. While infection rates are currently low, ZIKV remains a public health concern with no treatment or vaccine available. Targeting a cellular pathway to inhibit viral replication is a potential treatment strategy that avoids development of antiviral resistance. We demonstrate in this study that the non-toxic autophagy-inducing disaccharide trehalose reduces spread and output of ZIKV in infected neural progenitor cells (NPCs), the major cells infected in the fetus. We show that ZIKV spreads cell-to-cell in NPCs as an infectious center and that NPCs are more permissive to infection by contact with infected cells than by cell-free virus. We find that neutralizing antibody does not prevent the spread of the infection in NPCs. These results are significant in demonstrating anti-ZIKV activity of trehalose and in clarifying the primary means of Zika virus spread in clinically relevant target cells.

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Zika Virus: New Clinical Syndromes and Its Emergence in the Western Hemisphere

Journal of Virology ◽

10.1128/jvi.00252-16 ◽

2016 ◽

Vol 90 (10) ◽

pp. 4864-4875 ◽

Cited By ~ 279

Author(s):

Helen M. Lazear ◽

Michael S. Diamond

Keyword(s):

Zika Virus ◽

Antiviral Agents ◽

Western Hemisphere ◽

Health Concern ◽

Global Public Health ◽

Public Health Concern ◽

Zikv Infection ◽

Clinical Complications ◽

Clinical Syndromes ◽

Potential Interactions

Zika virus (ZIKV) had remained a relatively obscure flavivirus until a recent series of outbreaks accompanied by unexpectedly severe clinical complications brought this virus into the spotlight as causing an infection of global public health concern. In this review, we discuss the history and epidemiology of ZIKV infection, recent outbreaks in Oceania and the emergence of ZIKV in the Western Hemisphere, newly ascribed complications of ZIKV infection, including Guillain-Barré syndrome and microcephaly, potential interactions between ZIKV and dengue virus, and the prospects for the development of antiviral agents and vaccines.

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Resveratrol affects Zika virus replication in vitro

Scientific Reports ◽

10.1038/s41598-019-50674-3 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 10

Author(s):

Azirah Mohd ◽

Nurhafiza Zainal ◽

Kim-Kee Tan ◽

Sazaly AbuBakar

Keyword(s):

Cell Cultures ◽

Zika Virus ◽

Copy Number ◽

Virus Titer ◽

Health Concern ◽

Dependent Manner ◽

Zikv Infection ◽

Mrna Copy Number ◽

Short Period

Abstract Zika virus (ZIKV) infection is a serious public health concern. ZIKV infection has been associated with increased occurrences of microcephaly among newborns and incidences of Guillain-Barré syndrome among adults. No specific therapeutics or vaccines are currently available to treat and protect against ZIKV infection. Here, a plant-secreted phytoalexin, resveratrol (RES), was investigated for its ability to inhibit ZIKV replication in vitro. Several RES treatment regimens were used. The ZIKV titers of mock- and RES-treated infected cell cultures were determined using the focus-forming assay and the Zika mRNA copy number as determined using qRT-PCR. Our results suggested that RES treatment reduced ZIKV titers in a dose-dependent manner. A reduction of >90% of virus titer and ZIKV mRNA copy number was achieved when infected cells were treated with 80 µM of RES post-infection. Pre-incubation of the virus with 80 µM RES showed >30% reduction in ZIKV titers and ZIKV mRNA copy number, implying potential direct virucidal effects of RES against the virus. The RES treatment reduced >70% virus titer in the anti-adsorption assay, suggesting the possibility that RES also interferes with ZIKV binding. However, there was no significant decrease in ZIKV titer when a short-period of RES treatment was applied to cells before ZIKV infection (pre-infection) and after the virus bound to the cells (virus internalization inhibition), implying that RES acts through its continuous presence in the cell cultures after virus infection. Overall, our results suggested that RES exhibited direct virucidal activity against ZIKV and possessed anti-ZIKV replication properties, highlighting the need for further exploration of RES as a potential antiviral molecule against ZIKV infection.

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Syndromic Multisuture Craniosynostosis With Associated Anterior Segment Dysgenesis, Optic Nerve Hypoplasia, and Congenital Glaucoma

The Cleft Palate-Craniofacial Journal ◽

10.1177/1055665618820481 ◽

2018 ◽

Vol 56 (6) ◽

pp. 823-826

Author(s):

Kelly P. Schultz ◽

Claire J. Wiggins ◽

Haley Streff ◽

Veeral S. Shah ◽

Edward P. Buchanan

Keyword(s):

Optic Nerve ◽

Anterior Segment ◽

Optic Nerve Hypoplasia ◽

Congenital Glaucoma ◽

Anterior Segment Dysgenesis

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Neural progenitor cell pyroptosis contributes to Zika virus-induced brain atrophy and represents a therapeutic target

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2007773117 ◽

2020 ◽

Vol 117 (38) ◽

pp. 23869-23878 ◽

Cited By ~ 2

Author(s):

Zhenjian He ◽

Shu An ◽

Jiahui Chen ◽

Shuqing Zhang ◽

Chahui Tan ◽

...

Keyword(s):

Neural Development ◽

Zika Virus ◽

Brain Atrophy ◽

Progenitor Cell ◽

Inflammatory Responses ◽

Neural Progenitor Cell ◽

Neural Progenitor ◽

Zikv Infection ◽

Caspase 1

Mounting evidence has associated Zika virus (ZIKV) infection with congenital malformations, including microcephaly, which raises global alarm. Nonetheless, mechanisms by which ZIKV disrupts neurogenesis and causes microcephaly are far from being understood. In this study, we discovered direct effects of ZIKV on neural progenitor cell development by inducing caspase-1– and gasdermin D (GSDMD)-mediated pyroptotic cell death, linking ZIKV infection with the development of microcephaly. Importantly, caspase-1 depletion or its inhibitor VX-765 treatment reduced ZIKV-induced inflammatory responses and pyroptosis, and substantially attenuated neuropathology and brain atrophy in vivo. Collectively, our data identify caspase-1– and GSDMD-mediated pyroptosis in neural progenitor cells as a previously unrecognized mechanism for ZIKV-related pathological effects during neural development, and also provide treatment options for ZIKV-associated diseases.

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Coordination of Zika Virus Infection and Viroplasm Organization by Microtubules and Microtubule-Organizing Centers

Cells ◽

10.3390/cells10123335 ◽

2021 ◽

Vol 10 (12) ◽

pp. 3335

Author(s):

Rebecca A. Buchwalter ◽

Sarah C. Ogden ◽

Sara B. York ◽

Li Sun ◽

Chunfeng Zheng ◽

...

Keyword(s):

Zika Virus ◽

Infectious Virus ◽

Virus Production ◽

Health Concern ◽

Zikv Infection ◽

Virus Particles ◽

Microtubule Organizing Centers ◽

Infected Cells ◽

Congenital Microcephaly ◽

A Site

Zika virus (ZIKV) became a global health concern in 2016 due to its links to congenital microcephaly and other birth defects. Flaviviruses, including ZIKV, reorganize the endoplasmic reticulum (ER) to form a viroplasm, a compartment where virus particles are assembled. Microtubules (MTs) and microtubule-organizing centers (MTOCs) coordinate structural and trafficking functions in the cell, and MTs also support replication of flaviviruses. Here we investigated the roles of MTs and the cell’s MTOCs on ZIKV viroplasm organization and virus production. We show that a toroidal-shaped viroplasm forms upon ZIKV infection, and MTs are organized at the viroplasm core and surrounding the viroplasm. We show that MTs are necessary for viroplasm organization and impact infectious virus production. In addition, the centrosome and the Golgi MTOC are closely associated with the viroplasm, and the centrosome coordinates the organization of the ZIKV viroplasm toroidal structure. Surprisingly, viroplasm formation and virus production are not significantly impaired when infected cells have no centrosomes and impaired Golgi MTOC, and we show that MTs are anchored to the viroplasm surface in these cells. We propose that the viroplasm is a site of MT organization, and the MTs organized at the viroplasm are sufficient for efficient virus production.

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Zika virus dysregulates the expression of astrocytic genes involved in neurodevelopment

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0009362 ◽

2021 ◽

Vol 15 (4) ◽

pp. e0009362

Author(s):

Muhammad Adnan Shereen ◽

Nadia Bashir ◽

Rui Su ◽

Fang Liu ◽

Kailang Wu ◽

...

Keyword(s):

Brain Development ◽

Signaling Pathway ◽

Glial Cells ◽

Zika Virus ◽

Molecular Mechanisms ◽

French Polynesia ◽

Health Concern ◽

Hippo Signaling ◽

Sequencing Data ◽

Zikv Infection

Zika virus (ZIKV) is a kind of flavivirus emerged in French Polynesia and Brazil, and has led to a worldwide public health concern since 2016. ZIKV infection causes various neurological conditions, which are associated with fetus brain development or peripheral and central nervous systems (PNS/CNS) functional problems. To date, no vaccine or any specific antiviral therapy against ZIKV infection are available. It urgently needs efforts to explore the underlying molecular mechanisms of ZIKV-induced neural pathogenesis. ZIKV favorably infects neural and glial cells specifically astrocytes, consequently dysregulating gene expression and pathways with impairment of process neural cells. In this study, we applied a model for ZIKV replication in mouse primary astrocytes (MPAs) and profiled temporal alterations in the host transcriptomes upon ZIKV infection. Among the RNA-sequencing data of 27,812 genes, we examined 710 genes were significantly differentially expressed by ZIKV, which lead to dysregulation of numerous functions including neurons development and migration, glial cells differentiation, myelinations, astrocytes projection, neurogenesis, and brain development, along with multiple pathways including Hippo signaling pathway, tight junction, PI3K-Akt signaling pathway, and focal adhesion. Furthermore, we confirmed the dysregulation of the selected genes in MPAs and human astroglioma U251 cells. We found that PTBP1, LIF, GHR, and PTBP3 were upregulated while EDNRB and MBP were downregulated upon ZIKV infection. The current study highlights the ZIKV-mediated potential genes associated with neurodevelopment or related diseases.

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Cross-Reaction, Enhancement, and Neutralization Activity of Dengue Virus Antibodies against Zika Virus: A Study in the Mexican Population

Journal of Immunology Research ◽

10.1155/2019/7239347 ◽

2019 ◽

Vol 2019 ◽

pp. 1-14 ◽

Cited By ~ 7

Author(s):

Mayra R. Montecillo-Aguado ◽

Alfredo E. Montes-Gómez ◽

Julio García-Cordero ◽

Josselin Corzo-Gómez ◽

Héctor Vivanco-Cid ◽

...

Keyword(s):

Dengue Virus ◽

Acute Phase ◽

Zika Virus ◽

Neutralizing Antibodies ◽

Cross Reactivity ◽

Health Concern ◽

Serum Samples ◽

Zikv Infection ◽

Antibody Dependent Enhancement ◽

Homologous Antigen

Zika virus (ZIKV), an emerging mosquito-borne flavivirus, has quickly spread in many regions around the world where dengue virus (DENV) is endemic. This represents a major health concern, given the high homology between these two viruses, which can result in cross-reactivity. The aim of this study was to determine the cross-reacting antibody response of the IgM and IgG classes against the recombinant envelope protein of ZIKV (rE-ZIKV) in sera from patients with acute-phase infection of different clinical forms of dengue, i.e., dengue fever (DF) and dengue hemorrhagic fever (DHF) (before the arrival of ZIKV in Mexico 2010), as well as acute-phase sera of ZIKV patients, together with the implications in neutralization and antibody-dependent enhancement. Differences in IgM responses were observed in a number of DF and DHF patients whose sera cross-reacted with the rE-ZIK antigen, with 42% recognition between acute-phase DHF and ZIKV but 27% recognition between DF and ZIKV. Regarding IgG antibodies, 71.5% from the DF group showed cross-reactivity to rE-ZIKV in contrast with 50% and only 25% of DHF and ZIKV serum samples, respectively, which specifically recognized the homologous antigen. The DHF group showed more enhancement of ZIKV infection of FCRγ-expressing cells compared to the DF group. Furthermore, the DHF group also showed a higher cross-neutralizing ability than that of DF. This is the first report where DF and DHF serum samples were evaluated for cross-reactivity against Zika protein and ZIKV. Furthermore, DENV serum samples cross-protect against ZIKV through neutralizing antibodies but at the same time mediate antibody-dependent enhancement in the sequential ZIKV infection.

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A Chimeric Zika Virus between Viral Strains MR766 and BeH819015 Highlights a Role for E-glycan Loop in Antibody-mediated Virus Neutralization

Vaccines ◽

10.3390/vaccines7020055 ◽

2019 ◽

Vol 7 (2) ◽

pp. 55 ◽

Cited By ~ 10

Author(s):

Etienne Frumence ◽

Wildriss Viranaicken ◽

Sandra Bos ◽

Maria-Teresa Alvarez-Martinez ◽

Marjolaine Roche ◽

...

Keyword(s):

Zika Virus ◽

Protective Immunity ◽

Neutralizing Antibody ◽

Health Concern ◽

Zikv Infection ◽

Humoral Immune ◽

African Strain ◽

Antibody Titers ◽

Antibody Epitopes ◽

Viral Clone

Zika virus (ZIKV) is an emerging mosquito-borne flavivirus which is of major public health concern. ZIKV infection is recognized as the cause of congenital Zika disease and other neurological defects, with no specific prophylactic or therapeutic treatments. As the humoral immune response is an essential component of protective immunity, there is an urgent need for effective vaccines that confer protection against ZIKV infection. In the present study, we evaluate the immunogenicity of chimeric viral clone ZIKBeHMR-2, in which the region encoding the structural proteins of the African strain MR766 backbone was replaced with its counterpart from the epidemic strain BeH819015. Three amino-acid substitutions I152T, T156I, and H158Y were introduced in the glycan loop of the E protein (E-GL) making ZIKBeHMR-2 a non-glycosylated virus. Adult BALB/c mice inoculated intraperitoneally with ZIKBeHMR-2 developed anti-ZIKV antibodies directed against viral proteins E and NS1 and a booster dose increased antibody titers. Immunization with ZIKBeHMR-2 resulted in a rapid production of neutralizing anti-ZIKV antibodies. Antibody-mediated ZIKV neutralization was effective against viral strain MR766, whereas epidemic ZIKV strains were poorly sensitive to neutralization by anti-ZIKBeHMR-2 immune sera. From our data, we propose that the three E-GL residues at positions E-152, E-156, and E-158 greatly influence the accessibility of neutralizing antibody epitopes on ZIKV.

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