The Epithelial-to-Mesenchymal Transition-Like Process Induced by TGF-β1 Enhances Rubella Virus Binding and Infection in A549 Cells via the Smad Pathway

Pham, Ngan Thi Kim; Trinh, Quang Duy; Takada, Kazuko; Takano, Chika; Sasano, Mari; Okitsu, Shoko; Ushijima, Hiroshi; Komine‐Aizawa, Shihoko; Hayakawa, Satoshi

doi:10.3390/microorganisms9030662

Cited by 6 publications

(7 citation statements)

References 32 publications

(37 reference statements)

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“…To further confirm the functional significance and assess the targetability, we inhibited p-EMT activation in UM‑Chor1 cells and primary cultures of SBC cells by treatment with TGF-βR1 inhibitors (LY364947, Vactosertib, PF06952229, YL-13027) for 48 h 44 , 45 . The high (++) and low (+) concentrations of LY364947, Vactosertib, and YL13027 were 100 µM and 10 µM, respectively.…”

Section: Resultsmentioning

confidence: 99%

Single-cell transcriptome reveals cellular hierarchies and guides p-EMT-targeted trial in skull base chordoma

et al. 2022

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Skull base chordoma (SBC) is a bone cancer with a high recurrence rate, high radioresistance rate, and poorly understood mechanism. Here, we profiled the transcriptomes of 90,691 single cells, revealed the SBC cellular hierarchies, and explored novel treatment targets. We identified a cluster of stem-like SBC cells that tended to be distributed in the inferior part of the tumor. Combining radiated UM-Chor1 RNA-seq data and in vitro validation, we further found that this stem-like cell cluster is marked by cathepsin L (CTSL), a gene involved in the packaging of telomere ends, and may be responsible for radioresistance. Moreover, signatures related to partial epithelial–mesenchymal transition (p-EMT) were found to be significant in malignant cells and were related to the invasion and poor prognosis of SBC. Furthermore, YL-13027, a p-EMT inhibitor that acts through the TGF-β signaling pathway, demonstrated remarkable potency in inhibiting the invasiveness of SBC in preclinical models and was subsequently applied in a phase I clinical trial that enrolled three SBC patients. Encouragingly, YL-13027 attenuated the growth of SBC and achieved stable disease with no serious adverse events, underscoring the clinical potential for the precision treatment of SBC with this therapy. In summary, we conducted the first single-cell RNA sequencing of SBC and identified several targets that could be translated to the treatment of SBC.

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Section: Resultsmentioning

confidence: 99%

Single-cell transcriptome reveals cellular hierarchies and guides p-EMT-targeted trial in skull base chordoma

et al. 2022

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“…The cells were collected 24 hpi and subjected to intracellular staining of RuV capsid protein. The viral titer (in infectious units (IUs)) of a sample was calculated as the average of 3 titers measured in 3 consecutive wells with a percentage of RuV-infected cells lower than 40% and higher than 0.3%, as described previously [17,18].…”

Section: Viral Infectionmentioning

confidence: 99%

Low Susceptibility of Rubella Virus in First-Trimester Trophoblast Cell Lines

Pham

Trinh

Takada

et al. 2022

Viruses

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We recently published an article about myelin oligodendrocyte glycoprotein-independent rubella infection of keratinocytes in vitro, in which first-trimester trophoblast cells were shown as rubella virus (RuV)-resistant. Given an incident rate as high as 90% of congenital rubella syndrome in the first eight weeks of pregnancy, the RuV infection of first-trimester trophoblasts is considered key to opening the gate to transplacental transmission mechanisms. Therefore, with this study, we aimed to verify the susceptibility/resistance of first-trimester trophoblast cell lines, HTR-8/SVneo and Swan.71, against RuV. Cells cultured on multi-well plates were challenged with a RuV clinical strain at a multiplicity of infection from 5 to 10 for 3 h. The infectivity was investigated by immunofluorescence (IF) assay and flow cytometry (FCM) analysis. Supernatants collected during the post-infection period were used to determine virus-progeny production. The scattered signaling of RuV infection of these cells was noted by IF assay, and the FCM analysis showed an average of 4–5% of gated cells infected with RuV. In addition, a small but significant production of virus progeny was also observed. In conclusion, by employing appropriate approaches, we determined the low infectivity of RuV in first-trimester trophoblast cell lines but not resistance as in our previous report.

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“…The viral stock solution was prepared by propagating the virus in Vero cells and concentrating the viral particles via ultracentrifugation at 52,000 × g for 90 min in a Himac CS100GX microultracentrifuge with an S50A rotor (Hitachi Koki Co., Ltd., Ibaraki, Japan). Viral titers were estimated with the TCID50 method or flow cytometry (FCM) analysis, as described previously (Grigorov et al, 2011;Pham et al, 2021).…”

Section: Cell Culture and Virusmentioning

confidence: 99%

“…Viral titers [in infectious units (IUs)] were determined by FCM analysis. The viral titer in a sample was calculated as the average of 3 titers measured in 3 consecutive wells with a percentage of RuV-infected cells lower than 40% and higher than 0.3%, as described previously (Grigorov et al, 2011;Pham et al, 2021).…”

Section: Viral Infectionmentioning

confidence: 99%

Enhancement of Rubella Virus Infection in Immortalized Human First-Trimester Trophoblasts Under Low-Glucose Stress Conditions

et al. 2022

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Rubella virus (RuV) infections in pregnant women, especially first-trimester infections, can lead to congenital rubella syndrome (CRS). However, the mechanisms of fetal RuV infection are not completely understood, and it is not observed in every pregnant woman infected with RuV. As gestational diabetes mellitus is a risk factor for congenital viral infections, we investigated the possible roles of hypoglycemia-related endoplasmic reticulum (ER) stress as a key factor for vertical RuV infection using immortalized human first-trimester trophoblasts. Low-glucose stress was induced prior to RuV infection by culturing HTR-8/SVneo and Swan.71 cells in low-glucose (LG) medium for 24 h or high-glucose medium for 6 h and then LG medium for an additional 18 h. Clinically isolated RuV was inoculated at a multiplicity of infection of 5 to 10. The intracellular localization of the RuV capsid protein was investigated 24 to 48 h post-infection (pi) with flow cytometry (FCM) analysis and fluorescence microscopy. Viral progeny production was monitored by FCM analysis. Increases in RuV infection in LG-induced ER-stressed trophoblasts were observed. No significant increase in apoptosis of RuV-infected cells was noted at days 2 and 5 pi, and substantial viral progeny production was observed until day 5 pi. An approximate fivefold increase in viral binding was noted for the LG-stressed cells. Although the detailed mechanisms underlying viral entry into LG-stressed cells are not known and require further investigation, these findings suggest that a certain degree of LG stress in early pregnancy may facilitate infection and cause CRS.

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The Epithelial-to-Mesenchymal Transition-Like Process Induced by TGF-β1 Enhances Rubella Virus Binding and Infection in A549 Cells via the Smad Pathway

Cited by 6 publications

References 32 publications

Single-cell transcriptome reveals cellular hierarchies and guides p-EMT-targeted trial in skull base chordoma

Single-cell transcriptome reveals cellular hierarchies and guides p-EMT-targeted trial in skull base chordoma

Low Susceptibility of Rubella Virus in First-Trimester Trophoblast Cell Lines

Enhancement of Rubella Virus Infection in Immortalized Human First-Trimester Trophoblasts Under Low-Glucose Stress Conditions

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