Intrinsic Immune Mechanisms Restricting Human Cytomegalovirus Replication

Schilling, Eva-Maria; Scherer, Myriam; Stamminger, Thomas

doi:10.3390/v13020179

Cited by 14 publications

(11 citation statements)

References 101 publications

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“…to the HCMV genome, resulting in the formation of a repressive chromatin structure and epigenetic silencing of viral gene transcription. 6,29,30 To test whether PML is involved in MORC3-mediated MIEP…”

Section: Overexpressing Morc3 Suppresses Hcmv Replicationmentioning

confidence: 99%

MORC3 restricts human cytomegalovirus infection by suppressing the major immediate‐early promoter activity

Yao

Wang

et al. 2022

Journal of Medical Virology

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During the long coevolution of human cytomegalovirus (HCMV) and humans, the host has formed a defense system of multiple layers to eradicate the invader, and the virus has developed various strategies to evade host surveillance programs. The intrinsic immunity primarily orchestrated by promyelocytic leukemia (PML) nuclear bodies (PML-NBs) represents the first line of defense against HCMV infection. Here, we demonstrate that microrchidia family CW-type zinc finger 3 (MORC3), a PML-NBs component, is a restriction factor targeting HCMV infection. We show that depletion of MORC3 through knockdown by RNA interference or knockout by CRISPR-Cas9 augmented immediate-early protein 1 (IE1) gene expression and subsequent viral replication, and overexpressing MORC3 inhibited HCMV replication by suppressing IE1 gene expression. To relief the restriction, HCMV induces transient reduction of MORC3 protein level via the ubiquitin-proteasome pathway during the immediate-early to early stage. However, MORC3 transcription is upregulated, and the protein level recovers in the late stages. Further analyses with temporal-controlled MORC3 expression and the major immediate-early promoter (MIEP)-based reporters show that MORC3 suppresses MIEP activity and consequent IE1 expression with the assistance of PML. Taken together, our data reveal that HCMV enforces temporary loss of MORC3 to evade its repression against the initiation of immediate-early gene expression.

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Section: Overexpressing Morc3 Suppresses Hcmv Replicationmentioning

confidence: 99%

MORC3 restricts human cytomegalovirus infection by suppressing the major immediate‐early promoter activity

Yao

Wang

et al. 2022

Journal of Medical Virology

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“…This might be advantageous in the presence of antiviral host defense by reducing the time until viral immune evasion genes are expressed. Additionally, it is conceivable that such a high virus dose is necessary for HCMV in vivo to overcome cellular restriction factors that are upregulated by interferons [ 67 , 68 , 69 ]. Consistent with this, cell-associated spread has recently been reported to be more resistant to interferon-induced antiviral factors in comparison to cell-free spread [ 37 ].…”

Section: Discussionmentioning

confidence: 99%

Peptide Derivatives of Platelet-Derived Growth Factor Receptor Alpha Inhibit Cell-Associated Spread of Human Cytomegalovirus

Braun¹,

Fischer²,

Sampaio³

et al. 2021

Viruses

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Cell-free human cytomegalovirus (HCMV) can be inhibited by a soluble form of the cellular HCMV-receptor PDGFRα, resembling neutralization by antibodies. The cell-associated growth of recent HCMV isolates, however, is resistant against antibodies. We investigated whether PDGFRα-derivatives can inhibit this transmission mode. A protein containing the extracellular PDGFRα-domain and 40-mer peptides derived therefrom were tested regarding the inhibition of the cell-associated HCMV strain Merlin-pAL1502, hits were validated with recent isolates, and the most effective peptide was modified to increase its potency. The modified peptide was further analyzed regarding its mode of action on the virion level. While full-length PDGFRα failed to inhibit HCMV isolates, three peptides significantly reduced virus growth. A 30-mer version of the lead peptide (GD30) proved even more effective against the cell-free virus, and this effect was HCMV-specific and depended on the viral glycoprotein O. In cell-associated spread, GD30 reduced both the number of transferred particles and their penetration. This effect was reversible after peptide removal, which allowed the synchronized analysis of particle transfer, showing that two virions per hour were transferred to neighboring cells and one virion was sufficient for infection. In conclusion, PDGFRα-derived peptides are novel inhibitors of the cell-associated spread of HCMV and facilitate the investigation of this transmission mode.

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“…These restriction factors play various roles in the host immune response against CMV: Galectin-9 restricts entry of CMV into host cells, MXB interferes with the nuclear delivery of CMV genomes, PML nuclear bodies associate with CMV to induce epigenetic silencing, and BCL2L14 restricts CMV viral replication via regulation of the type I interferon response. 13 RSAD2 codes for the cytoplasmic antiviral protein viperin that is directly induced by CMV. 14 Our results thus provide the first human data on the involvement of these specific restriction factors in congenital CMV infection.…”

Section: Comment Principal Findingsmentioning

confidence: 99%

RNAseq of amniotic fluid cell-free RNA: insights into the pathophysiology of congenital cytomegalovirus infection

Hui

Catte

Beard

et al. 2022

Preprint

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Background Congenital cytomegalovirus (cCMV) is the most common perinatal infection and a significant cause of sensorineural hearing loss, cerebral palsy and neurodevelopmental disability. There is a paucity of human gene expression studies examining the pathophysiology of CMV infection. Objectives The aim of this study was to perform a whole transcriptomic assessment of amniotic fluid from pregnancies with live fetuses to identify differentially-expressed genes and enriched Gene Ontology categories associated with cCMV infection. Study design Amniotic fluid supernatant was prospectively collected from pregnant women undergoing amniocentesis for suspected cCMV due to first trimester maternal primary infection or ultrasound features suggestive of fetal infection. Women who had received therapy to prevent fetal infection were excluded. cCMV was diagnosed via viral PCR of amniotic fluid; CMV-infected fetuses were paired with noninfected controls, matched for gestational age and fetal sex. Paired-end RNA sequencing was performed on amniotic fluid cell-free RNA with the Novaseq6000 at a depth of 30 million reads/sample. Following quality control and filtering, reads were mapped to the human genome and counts summarised across genes. Differentially expressed genes were identified using two approaches: voomWithQualityWeights in conjunction with limma and RUVSeq with edgeR. Genes with a false discovery rate (FDR) < 0.05 were considered statistically significant. Differential exon usage was analysed using DEXSeq. Functional analysis was performed using Gene Set Enrichment Analysis and Ingenuity Pathway Analysis. Manual curation of differentially regulated genes was also performed. Results Amniotic fluid samples were collected from 50 women; 16 (32%) had cCMV confirmed by PCR. After excluding 3 samples without matched controls, 13 CMV-infected samples collected at 18-23 weeks and 13 CMV-negative gestation matched controls were submitted for RNA sequencing and analysis (total n=26). Ten of the 13 pregnancies with CMV-infected fetuses had amniocentesis due to serological evidence of maternal primary infection with normal fetal ultrasound, and three had amniocentesis due to ultrasound abnormality suggestive of CMV infection. Four CMV-infected pregnancies ended in termination (n=3) or fetal death (n=1), and 9 resulted in live births. Pregnancy outcomes were available on 11 of the 13 CMV-negative controls; all resulted in live births of clinically-well infants. Differential gene expression analysis revealed 309 up-regulated and 32 down-regulated genes in the CMV-infected group compared with the CMV-negative group. Gene set enrichment analysis showed significant enrichment of multiple Gene Ontology categories involving the innate immune response to viral infection and interferon signalling. Of the 32 significantly down-regulated genes, 8 were known to be involved in neurodevelopment and preferentially expressed by the brain. Six specific cellular restriction factors involved in host defence to CMV infection were upregulated in the CMV-infected group. Ingenuity Pathway Analysis predicted activation of pathways involved in progressive neurological disease, and inflammatory neurological disease. Conclusions In this next generation sequencing study, we reveal new insights into the pathophysiology of cCMV infection. These data on the upregulation of the intraamniotic innate immune response to CMV infection and the dysregulation of neurodevelopmental genes may inform future approaches to developing prognostic markers and assessing fetal responses to in utero therapy.

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Intrinsic Immune Mechanisms Restricting Human Cytomegalovirus Replication

Cited by 14 publications

References 101 publications

MORC3 restricts human cytomegalovirus infection by suppressing the major immediate‐early promoter activity

MORC3 restricts human cytomegalovirus infection by suppressing the major immediate‐early promoter activity

Peptide Derivatives of Platelet-Derived Growth Factor Receptor Alpha Inhibit Cell-Associated Spread of Human Cytomegalovirus

RNAseq of amniotic fluid cell-free RNA: insights into the pathophysiology of congenital cytomegalovirus infection

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