Transcriptome profiling highlights regulated biological processes and type III interferon antiviral responses upon Crimean-Congo hemorrhagic fever virus infection

Mo, Qiong; Feng, Kuan; Dai, Shiyu; Wu, Qiaoli; Zhang, Zhong; Ali, Ashaq; Deng, Fei; Wang, Hualin; Ning, Yun‐Jia

doi:10.1016/j.virs.2022.09.002

Cited by 4 publications

(5 citation statements)

References 75 publications

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“…Besides, unassembled glycoproteins can be deployed to ubiquitylation and proteasomal degradation from ER by the ERAD pathway 43 . In our study, some Gn/Gc-interacting chaperone proteins are involved in ER stress and ERAD pathway, including members of HSP70 family (HSPA5, HSPA8 and HSPA1B), SSR4, THBS4 and UGGT1, which is consistent with ER stress caused by CCHFV infection as reported previously by us and others 44,45 . The data obtained here may provide clues and host targets for further depiction of the molecular mechanisms underlying CCHFV-infection-triggered stress responses.…”

Section: Hax1 Inhibits Cchfv Gp-mediated Virion Packaging and Acts As...supporting

confidence: 92%

“…The HAX1 RNAi constructs were generated by cloning double-stranded oligonucleotides targeting specific sequences into the shRNA-expressing vector, pLKO.1 as previously described 45,57 . The target sequences of each shRNA plasmid were listed in Table S3.…”

Section: Gene Silencingmentioning

confidence: 99%

“…The single-guide RNA (sgRNA) sequence targeting exo2 of HAX1 was designed with the online CRISPR Design tool (http://crispor.tefor.net/) and cloned into pSpCas9(BB)-2A-Puro (PX459) V2.0 vector as previously described 45,57 . HEK293 cells were transfected with the constructed pX459-based plasmids for 48 h and selected with 3 μg/mL puromycin (Gibco, Cat#A1113803) for 72 h. Then single clonal cell lines were isolated from the transfected cells by serial dilution, and identified by gene sequencing and Western blot analysis.…”

Section: Generation Of Hax1-ko Cell Lines By Crispr-cas9mentioning

confidence: 99%

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Interactome analysis of Crimean-Congo hemorrhagic fever virus glycoproteins reveals HAX1 as a host restriction factor against the virion packaging

Dai

Feng

et al. 2023

Preprint

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Crimean-Congo hemorrhagic fever virus (CCHFV) is a biosafety level-4 and World Health Organization-recognized priority pathogen requiring urgent Research & Development attention. The glycoproteins of CCHFV, Gn and Gc, are considered to play multiple roles in the viral life cycle by interactions with host cells; however, these interactions remain largely unclear to date. Here, we analyzed the cellular interactomes of CCHFV glycoproteins by affinity purification-quantitative mass spectrometry and identified 45 host proteins as high-confidence Gn/Gc interactors. These host molecules are involved in transmembrane transport, metabolism, oxidative stress, protein folding and glycosylation, etc., in line with the potential physiological actions of the viral glycoproteins. As a proof of principle, we detailedly elucidated the role of an identified cellular protein, HAX1, and therefore uncovered an interesting host antiviral strategy. HAX1 interacts with Gn by its C-terminus, while its N-terminal region leads to mitochondrial localization. By the strong interaction, HAX1 sequestrates Gn to mitochondria, thus depriving Gn of its normal Golgi localization that is indispensable for functional glycoprotein-mediated progeny virion packaging. Consistently, the notable inhibitory activity of HAX1 against viral packaging and hence propagation was further elucidated in the contexts of pseudotyped and authentic CCHFV infections by series of loss/gain-of-function assays in cellular and animal infection models. Together, the findings not only provide a systematic CCHFV Gn/Gc-host protein-protein interaction map, but also unravel a novel cellular antiviral mechanism, which may facilitate further studies on CCHFV biology and therapeutic approaches.

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Section: Hax1 Inhibits Cchfv Gp-mediated Virion Packaging and Acts As...supporting

confidence: 92%

Section: Gene Silencingmentioning

confidence: 99%

Section: Generation Of Hax1-ko Cell Lines By Crispr-cas9mentioning

confidence: 99%

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Interactome analysis of Crimean-Congo hemorrhagic fever virus glycoproteins reveals HAX1 as a host restriction factor against the virion packaging

Dai

Feng

et al. 2023

Preprint

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“…Our earlier study reported system-level host's metabolic reprogramming toward CCM with distinct upregulation of OXPHOS during acute CCHFV infection ( 8 ). This corroborated with the transcriptomics or proteomics performed in vitro infection models in cancer cell lines ( 13 ) and in vivo infection studies in nonhuman primates ( 14 ) and mice ( 15 ), showing dysregulation of the metabolic process and upregulation in the IFN pathways. The temporal quantitative proteomics analysis by liquid chromatography–mass spectrometry (LC–MS) in the CCHFV-infected human hepatocellular carcinoma cell line, Huh7 model reported by us showed dynamic changes in the ISGs and metabolic process during the progressive replication ( 4 ).…”

Section: Discussionsupporting

confidence: 71%

Systems-level temporal immune-metabolic profile in Crimean-Congo hemorrhagic fever virus infection

Ambikan,

Elaldi,

Svensson-Akusjärvi

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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Crimean-Congo hemorrhagic fever (CCHF) caused by CCHF virus (CCHFV) is one of the epidemic-prone diseases prioritized by the World Health Organisation as public health emergency with an urgent need for accelerated research. The trajectory of host response against CCHFV is multifarious and remains unknown. Here, we reported the temporal spectrum of pathogenesis following the CCHFV infection using genome-wide blood transcriptomics analysis followed by advanced systems biology analysis, temporal immune-pathogenic alterations, and context-specific progressive and postinfection genome-scale metabolic models (GSMM) on samples collected during the acute (T0), early convalescent (T1), and convalescent-phase (T2). The interplay between the retinoic acid-inducible gene-I-like/nucleotide-binding oligomerization domain-like receptor and tumor necrosis factor signaling governed the trajectory of antiviral immune responses. The rearrangement of intracellular metabolic fluxes toward the amino acid metabolism and metabolic shift toward oxidative phosphorylation and fatty acid oxidation during acute CCHFV infection determine the pathogenicity. The upregulation of the tricarboxylic acid cycle during CCHFV infection, compared to the noninfected healthy control and between the severity groups, indicated an increased energy demand and cellular stress. The upregulation of glycolysis and pyruvate metabolism potentiated energy generation through alternative pathways associated with the severity of the infection. The downregulation of metabolic processes at the convalescent phase identified by blood cell transcriptomics and single-cell type proteomics of five immune cells (CD4 + and CD8 + T cells, CD14 + monocytes, B cells, and NK cells) potentially leads to metabolic rewiring through the recovery due to hyperactivity during the acute phase leading to post-viral fatigue syndrome.

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“…The use of a low MOI was intended to minimize a potential inhibitory effect by interferon carryover on the initiation and establishment of infection. Interferon responses are a known antiviral mechanism against CCHFV infection [ 15 , 47 , 52 , 79 – 82 ] and these responses have been reported in Vero and SW-13 cells [ 15 ]. Pre-treatment (24 h or 2 h before the infection) or early treatment (1 h post-infection) but not delayed treatment (6 h post-infection) of cell cultures with interferon was previously found to result in reduced viral titers, suggesting that the early stage of CCHFV infection in the cell culture context is more sensitive to interferon responses and the sensitivity drops once the virus is replicating [ 79 ].…”

Section: Resultsmentioning

confidence: 99%

Comparative characterization of Crimean-Congo hemorrhagic fever virus cell culture systems with application to propagation and titration methods

Smith

Goolia

et al. 2023

Virol J

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Crimean-Congo hemorrhagic fever orthonairovirus (CCHFV) is a biosafety level 4 and World Health Organization top priority pathogen. Infection leads to an often fatal hemorrhagic fever disease in humans. The tick-borne virus is endemic in countries across Asia, Europe and Africa, with signs of spreading into new regions. Despite the severity of disease and the potential of CCHFV geographic expansion to cause widespread outbreaks, no approved vaccine or treatment is currently available. Critical for basic research and the development of diagnostics or medical countermeasures, CCHFV viral stocks are commonly produced in Vero E6 and SW-13 cell lines. While a variety of in-house methods are being used across different laboratories, there has been no clear, specific consensus on a standard, optimal system for CCHFV growth and titration. In this study, we perform a systematic, side-by-side characterization of Vero E6 and SW-13 cell lines concerning the replication kinetics of CCHFV under different culture conditions. SW-13 cells are typically cultured in a CO2-free condition (SW-13 CO2−) according to the American Type Culture Collection. However, we identify a CO2-compatible culture condition (SW-13 CO2+) that demonstrates the highest viral load (RNA concentration) and titer (infectious virus concentration) in the culture supernatants, in comparison to SW-13 CO2− and Vero E6 cultures. This optimal viral propagation system also leads to the development of two titration methods: an immunostaining-based plaque assay using a commercial CCHFV antibody and a colorimetric readout, and an antibody staining-free, cytopathic effect-based median tissue culture infectious dose assay using a simple excel calculator. These are anticipated to serve as a basis for a reproducible, standardized and user-friendly platform for CCHFV propagation and titration.

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Transcriptome profiling highlights regulated biological processes and type III interferon antiviral responses upon Crimean-Congo hemorrhagic fever virus infection

Cited by 4 publications

References 75 publications

Interactome analysis of Crimean-Congo hemorrhagic fever virus glycoproteins reveals HAX1 as a host restriction factor against the virion packaging

Interactome analysis of Crimean-Congo hemorrhagic fever virus glycoproteins reveals HAX1 as a host restriction factor against the virion packaging

Systems-level temporal immune-metabolic profile in Crimean-Congo hemorrhagic fever virus infection

Comparative characterization of Crimean-Congo hemorrhagic fever virus cell culture systems with application to propagation and titration methods

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