mRNA and miRNA profiling of Zika virus-infected human umbilical cord mesenchymal stem cells identifies miR-142-5p as an antiviral factor

Abstract

Zika virus (ZIKV) infection during pregnancy is associated with congenital brain abnormalities, a finding that highlights the urgent need to understand mother-to-fetus transmission mechanisms. Human umbilical cord mesenchymal stem cells (hUCMSCs) are susceptible to ZIKV infection but the underlying mechanisms of viral susceptibility remain largely unexplored. In this study, we have characterized and compared host mRNA and miRNA expression profiles in hUCMSCs after infection with two lineages of ZIKV, African (MR766) and Asian (PRVABC59). RNA sequencing analysis identified differentially expressed genes involved in anti-viral immunity and mitochondrial dynamics following ZIKV infection. In particular, ZIKV-infected hUCMSCs displayed mitochondrial elongation and the treatment of hUCMSCs with mitochondrial fission inhibitor led to a dose-dependent increase in ZIKV gene expression and decrease in antiviral signalling pathways. Moreover, small RNA sequencing analysis identified several significantly up-or downregulated microRNAs. Interestingly, miR-142-5p was significantly downregulated upon ZIKV infection, whereas cellular targets of miR-142-5p, IL6ST and ITGAV, were upregulated. Overexpression of miR-142-5p resulted in the suppression of ZIKV replication. Furthermore, blocking ITGAV expression resulted in a significant suppression of ZIKV binding to cells, suggesting a potential role of ITGAV in ZIKV entry. In conclusion, these results demonstrate both common and specific host responses to African and Asian ZIKV lineages and indicate miR-142-5p as a key regulator of ZIKV replication in the umbilical cords.

SARS-CoV-2 Nucleocapsid Protein Targets RIG-I-Like Receptor Pathways to Inhibit the Induction of Interferon Response

“…A standard plaque assay was performed to determine viral titers in MDCK cells [ 13 , 14 ]. ZIKV MR766 (African lineage) was purchased from ATCC and propagated in Vero cells, as previously described [ 15 , 16 , 17 ].…”

“…Mito-dsRED plasmid (a kind gift of Dr. Woong Sun, Korea University School of Medicine) was used to indicate mitochondria. Cells were washed with phosphate-buffered saline (PBS), fixed in 4% paraformaldehyde, and permeabilized with 0.1% Triton X-100 as described previously [ 17 ]. Cells were then blocked and incubated with the indicated primary antibodies followed by incubation with a fluorescent secondary antibody.…”

Zika Virus Induces Tumor Necrosis Factor-Related Apoptosis Inducing Ligand (TRAIL)-Mediated Apoptosis in Human Neural Progenitor Cells

“…ZIKV MR766 (African lineage) and PRVABC59 (Asian lineage) strains were purchased from ATCC and propagated in Vero cells. Viral titers were determined using a standard plaque assay as described previously [ 27 ]. Viral stocks were aliquoted and stored at −80 °C until use.…”

“…Total RNA (0.5 µg) was isolated with Trizol reagent (Invitrogen) and reverse transcribed to generate cDNA using an ImProm-II Reverse Transcription System (Promega, Madison, WI, USA) for 1 h at 42 • C. The resulting cDNA was used as a template for qRT-PCR quantification of ZIKV and host transcript levels using a Power SYBR Green PCR Master Mix (Thermo Fisher Scientific, Waltham, MA, USA). The primer sequences used in this study are listed in Table 1 [27]. Quantification was carried out on a QuantStudio 6 Flex Real-time PCR system (Thermo Fisher Scientific, Waltham, MA, USA) for cDNA amplification with Power SYBR ® Green Master Mix (Invitrogen) under the following conditions: 95°C for 10 min, followed by 40 cycles of 95°C for 30 s, and 60°C for 1 min.…”

Identifying the functions of two biomarkers in human oligodendrocyte progenitor cell development

“…After obtaining the target cells, we studied the gene levels of these cells. Single-cell RNA sequencing (scRNA-seq) can be used to study the different subgroups of hOPCs before sorting [ 15 , 16 ], while RNA sequencing (RNA-seq) can be used to perform overall differential gene expression analysis and functional enrichment analysis on the sorted cell population [ 17 , 18 ]. In vivo and in vitro cell function is assessed mainly through in vitro proliferation [ 19 ] and migration experiments [ 20 ] as well as the evaluation of the myelinating ability of the sorted cells transplanted into the shiverer mouse corpus callosum [ 21 , 22 ].…”