Zika Virus Infection Prevents Host mRNA Nuclear Export by Disrupting UPF1 Function

AbstractZika virus (ZIKV) is a mosquito-borne RNA virus that can infect fetuses in utero causing characteristic neurodevelopmental disorders including microcephaly. We previously showed that ZIKV infection downregulates expression of up-frameshift protein 1 (UPF1), a helicase/ATPase and central regulator of the nonsense-mediated mRNA decay pathway. Here, we identify a novel function of nuclear UPF1 in mRNA export. Using crosslinking immunoprecipitation of UPF1 followed by sequencing of associated transcripts as well as fluorescence in situ hybridization experiments, we find widespread mRNA accumulation in the nucleus of human neural progenitor cells (NPCs) upon ZIKV infection or UPF1 knockdown. Knockdown of FREM2, a top UPF1 target transcript encoding an extra-cellular matrix protein critical in fetal development, decreased expression of pluripotency markers and increased expressed neuronal differentiation in NPCs, consistent with the model that trapping FREM2 mRNA in the nucleus perturbs proper NPC function. Collectively, our data uncover a new posttranscriptional mechanism by which ZIKV “shuts off” host mRNA export via UPF1. As we find UPF1 linked to many neurodevelopment pathways, we propose that the lack of host mRNA export contributes to the neurodevelopmental defects associated with ZIKV infection.

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Zika Virus Infection Induces DNA Damage Response in Human Neural Progenitors That Enhances Viral Replication

Journal of Virology ◽

10.1128/jvi.00638-19 ◽

2019 ◽

Vol 93 (20) ◽

Cited By ~ 10

Author(s):

Christy Hammack ◽

Sarah C. Ogden ◽

Joseph C. Madden ◽

Angelica Medina ◽

Chongchong Xu ◽

...

Keyword(s):

Cell Cycle ◽

Dna Damage ◽

Dna Replication ◽

Zika Virus ◽

Neural Progenitor Cells ◽

S Phase ◽

Zikv Infection ◽

Phase Arrest ◽

Damage Response ◽

Human Neural Progenitor Cells

ABSTRACT Zika virus (ZIKV) infection attenuates the growth of human neural progenitor cells (hNPCs). As these hNPCs generate the cortical neurons during early brain development, the ZIKV-mediated growth retardation potentially contributes to the neurodevelopmental defects of the congenital Zika syndrome. Here, we investigate the mechanism by which ZIKV manipulates the cell cycle in hNPCs and the functional consequence of cell cycle perturbation on the replication of ZIKV and related flaviviruses. We demonstrate that ZIKV, but not dengue virus (DENV), induces DNA double-strand breaks (DSBs), triggering the DNA damage response through the ATM/Chk2 signaling pathway while suppressing the ATR/Chk1 signaling pathway. Furthermore, ZIKV infection impedes the progression of cells through S phase, thereby preventing the completion of host DNA replication. Recapitulation of the S-phase arrest state with inhibitors led to an increase in ZIKV replication, but not of West Nile virus or DENV. Our data identify ZIKV’s ability to induce DSBs and suppress host DNA replication, which results in a cellular environment favorable for its replication. IMPORTANCE Clinically, Zika virus (ZIKV) infection can lead to developmental defects in the cortex of the fetal brain. How ZIKV triggers this event in developing neural cells is not well understood at a molecular level and likely requires many contributing factors. ZIKV efficiently infects human neural progenitor cells (hNPCs) and leads to growth arrest of these cells, which are critical for brain development. Here, we demonstrate that infection with ZIKV, but not dengue virus, disrupts the cell cycle of hNPCs by halting DNA replication during S phase and inducing DNA damage. We further show that ZIKV infection activates the ATM/Chk2 checkpoint but prevents the activation of another checkpoint, the ATR/Chk1 pathway. These results unravel an intriguing mechanism by which an RNA virus interrupts host DNA replication. Finally, by mimicking virus-induced S-phase arrest, we show that ZIKV manipulates the cell cycle to benefit viral replication.

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A global lipid map defines a network essential for Zika virus replication

10.1101/2020.01.27.910919 ◽

2020 ◽

Author(s):

Hans C. Leier ◽

Jules B. Weinstein ◽

Jennifer E. Kyle ◽

Joon-Yong Lee ◽

Lisa M. Bramer ◽

...

Keyword(s):

Zika Virus ◽

Neural Progenitor Cells ◽

Critical Role ◽

Ectopic Expression ◽

Cell Types ◽

Zikv Infection ◽

Human Neural Progenitor Cells ◽

Replication Sites ◽

Global Concern ◽

Sphingolipid Biosynthesis

AbstractZika virus (ZIKV), an arbovirus of global concern, remodels intracellular membranes to form replication sites. How ZIKV dysregulates lipid networks to allow this, and consequences for disease, is poorly understood. Here, we performed comprehensive lipidomics to create a lipid network map during ZIKV infection. We found that ZIKV significantly alters host lipid composition, with the most striking changes seen within subclasses of sphingolipids. Ectopic expression of ZIKV NS4B protein resulted in similar changes, demonstrating a role for NS4B in modulating sphingolipid pathways. Disruption of sphingolipid biosynthesis in various cell types, including human neural progenitor cells, blocked ZIKV infection. Additionally, the sphingolipid ceramide redistributes to ZIKV replication sites and increasing ceramide levels by multiple pathways sensitizes cells to ZIKV infection. Thus, we identify a sphingolipid metabolic network with a critical role in ZIKV replication and show that ceramide flux is a key mediator of ZIKV infection.

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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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Molecular signatures associated with ZIKV exposure in human cortical neural progenitors

10.1101/071183 ◽

2016 ◽

Author(s):

Feiran Zhang ◽

Christy Hammack ◽

Sarah C. Ogden ◽

Yichen Cheng ◽

Emily M. Lee ◽

...

Keyword(s):

Neural Progenitor Cells ◽

Expression Profiles ◽

Molecular Signatures ◽

Zikv Infection ◽

Host Interactions ◽

Expression Of Genes ◽

Dna Replication And Repair ◽

Human Neural Progenitor Cells ◽

Induced Apoptosis ◽

And Function

AbstractZika virus (ZIKV) infection causes microcephaly and has been linked to other brain abnormalities. How ZIKV impairs brain development and function is unclear. Here we systematically profiled transcriptomes of human neural progenitor cells exposed to Asian ZIKVC, African ZIKVM, and dengue virus (DENV). In contrast to the robust global transcriptome changes induced by DENV, ZIKV has a more selective and larger impact on expression of genes involved in DNA replication and repair. While overall expression profiles are similar, ZIKVC, but not ZIKVM, induces upregulation of viral response genes and TP53. P53 inhibitors can block the apoptosis induced by both ZIKVC and ZIKVM in hNPCs, with higher potency against ZIKVC-induced apoptosis. Our analyses reveal virus- and strain-specific molecular signatures associated with ZIKV infection. These datasets will help to investigate ZIKV-host interactions and identify neurovirulence determinants of ZIKV.

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Zika virus induces mitotic catastrophe in human neural progenitors by triggering unscheduled mitotic entry in the presence of DNA damage while functionally depleting nuclear PNKP

10.1101/2021.08.27.458001 ◽

2021 ◽

Author(s):

Malgorzata Rychlowska ◽

Abigail Agyapong ◽

Michael Weinfeld ◽

Luis M Schang

Keyword(s):

Dna Damage ◽

Zika Virus ◽

Cortical Neurons ◽

Genome Stability ◽

Neural Progenitor Cells ◽

Cellular Level ◽

Mitotic Catastrophe ◽

Zikv Infection ◽

Mitotic Entry ◽

Checkpoint Kinases

Vertical transmission of Zika virus (ZIKV) leads with high frequency to congenital ZIKV syndrome (CZS), whose worse outcome is microcephaly. However, the mechanisms of congenital ZIKV neurodevelopmental pathologies, including direct cytotoxicity to neural progenitor cells (NPC), placental insufficiency, and immune responses, remain incompletely understood. At the cellular level, microcephaly typically results from death or insufficient proliferation of NPC or cortical neurons. NPCs replicate fast, requiring efficient DNA damage responses to ensure genome stability. Like congenital ZIKV infection, mutations in the polynucleotide 5’-kinase 3’-phosphatase (PNKP) gene, which encodes a critical DNA damage repair enzyme, results in recessive syndromes often characterized by congenital microcephaly with seizures (MCSZ). We thus tested whether there were any links between ZIKV and PNKP. Here we show that a PNKP phosphatase inhibitor inhibits ZIKV replication. PNKP relocalized from the nucleus to the cytoplasm in infected cells, co-localizing with the marker of ZIKV replication factories (RF) NS1 and resulting in functional nuclear PNKP depletion. Although infected NPC accumulated DNA damage, they failed to activate the DNA damage checkpoint kinases Chk1 and Chk2. ZIKV also induced activation of cytoplasmic CycA/CDK1 complexes, which trigger unscheduled mitotic entry. Inhibition of CDK1 activity inhibited ZIKV replication and the formation of RF, supporting a role of cytoplasmic CycA/CDK1 in RF morphogenesis. In brief, ZIKV infection induces mitotic catastrophe resulting from unscheduled mitotic entry in the presence of DNA damage. PNKP and CycA/CDK1 are thus host factors participating in ZIKV replication in NPC, and probably pathogenesis.

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Zika virus alters DNA methylation status of genes involved in Hippo signaling pathway in human neural progenitor cells

Epigenomics ◽

10.2217/epi-2018-0180 ◽

2019 ◽

Vol 11 (10) ◽

pp. 1143-1161 ◽

Cited By ~ 5

Author(s):

Deepika Kandilya ◽

Silambarasan Maskomani ◽

Sukanya Shyamasundar ◽

Paul Anantharajah Tambyah ◽

Chan Shiao Yng ◽

...

Keyword(s):

Dna Methylation ◽

Progenitor Cells ◽

Signaling Pathway ◽

Zika Virus ◽

Neural Progenitor Cells ◽

Methylation Status ◽

Neural Progenitor ◽

Hippo Signaling ◽

Hippo Signaling Pathway ◽

Human Neural Progenitor Cells

Aim: This study was aimed to understand if Zika virus (ZIKV) alters the DNA methylome of human neural progenitor cells (hNPCs). Materials & methods: Whole genome DNA methylation profiling was performed using human methylationEPIC array in control and ZIKV infected hNPCs. Results & conclusion: ZIKV infection altered the DNA methylation of several genes such as WWTR1 (TAZ) and RASSF1 of Hippo signaling pathway which regulates organ size during brain development, and decreased the expression of several centrosomal-related microcephaly genes, and genes involved in stemness and differentiation in human neural progenitor cells. Overall, ZIKV downregulated the Hippo signaling pathway genes which perturb the stemness and differentiation process in hNPCs, which could form the basis for ZIKV-induced microcephaly.

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Correction to: ZIKV infection effects changes in gene splicing, isoform composition and lncRNA expression in human neural progenitor cells

Virology Journal ◽

10.1186/s12985-019-1122-z ◽

2019 ◽

Vol 16 (1) ◽

Cited By ~ 1

Author(s):

Benxia Hu ◽

Yongxia Huo ◽

Liping Yang ◽

Guijun Chen ◽

Minhua Luo ◽

...

Keyword(s):

Progenitor Cells ◽

Neural Progenitor Cells ◽

Neural Progenitor ◽

Zikv Infection ◽

Gene Splicing ◽

Lncrna Expression ◽

Human Neural Progenitor Cells

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In Vitro Zika Virus Infection of Human Neural Progenitor Cells: Meta-Analysis of RNA-Seq Assays

Microorganisms ◽

10.3390/microorganisms8020270 ◽

2020 ◽

Vol 8 (2) ◽

pp. 270 ◽

Cited By ~ 3

Author(s):

Rossella Gratton ◽

Paola Maura Tricarico ◽

Almerinda Agrelli ◽

Heverton Valentim Colaço da Silva ◽

Lucas Coêlho Bernardo ◽

...

Keyword(s):

Progenitor Cells ◽

Zika Virus ◽

Neural Progenitor Cells ◽

Meta Analysis ◽

Fetal Brain ◽

Neural Progenitor ◽

Gene Expression Omnibus ◽

Human Neural Progenitor Cells ◽

Apoptosis Pathways

The Zika virus (ZIKV) is an emergent arthropod-borne virus (arbovirus) responsible for congenital Zika syndrome (CZS) and a range of other congenital malformations. Evidence shows that ZIKV infects human neural progenitor cells (hNPCs) in the fetal brain, prompting inflammation and tissue damage/loss. Despite recent advances, little is known about the pathways involved in CZS pathogenesis. We performed a meta-analysis, gene ontology (GO), and pathway analysis of whole transcriptome studies with the aim of clarifying the genes and pathways potentially altered during hNPCs infection with ZIKV. We selected three studies (17 samples of infected hPNCs compared to hPNCs uninfected controls) through a systematic search of the Gene Expression Omnibus (GEO) database. The raw reads were trimmed, counted, and normalized. Next, we performed a rank product meta-analysis to detect consistently differentially expressed genes (DEGs) in these independent experiments. We detected 13 statistically significant DEGs. GO ontology and reactome analysis showed an enrichment of interferon, pro-inflammatory, and chemokines signaling and apoptosis pathways in ZIKV-infected cells. Moreover, we detected three possible new candidate genes involved in hNPCs infection: APOL6, XAF1, and TNFRSF1. Our results confirm that interferon (IFN) signaling dominates the ZIKV response, and that a crucial contribution is given by apoptotic pathways, which might elicit the CZS phenotype.

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An Update on Sexual Transmission of Zika Virus

Pathogens ◽

10.3390/pathogens7030066 ◽

2018 ◽

Vol 7 (3) ◽

pp. 66 ◽

Cited By ~ 18

Author(s):

Hercules Sakkas ◽

Petros Bozidis ◽

Xenofon Giannakopoulos ◽

Nikolaos Sofikitis ◽

Chrissanthy Papadopoulou

Keyword(s):

Central Nervous System ◽

Nervous System ◽

Reproductive System ◽

Zika Virus ◽

Rna Virus ◽

Sexual Transmission ◽

Limited Disease ◽

Zikv Infection ◽

Blood Sucking

Zika virus (ZIKV) is a single-stranded RNA virus belonging to the arthropod-borne flaviviruses (arboviruses) which are mainly transmitted by blood-sucking mosquitoes of the genus Aedes. ZIKV infection has been known to be rather asymptomatic or presented as febrile self-limited disease; however, during the last decade the manifestation of ZIKV infection has been associated with a variety of neuroimmunological disorders including Guillain–Barré syndrome, microcephaly and other central nervous system abnormalities. More recently, there is accumulating evidence about sexual transmission of ZIKV, a trait that has never been observed in any other mosquito-borne flavivirus before. This article reviews the latest information regarding the latter and emerging role of ZIKV, focusing on the consequences of ZIKV infection on the male reproductive system and the epidemiology of human-to-human sexual transmission.

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