Biology subjects

Power, U. F.

Publications and source records attributed to Power, U. F..

9 recordsLinked to original sources

bioRxiv2026

A host ATPase essential for rhinovirus replication is an antiviral target with a high barrier to resistance

Rhinoviruses are the leading cause of acute respiratory illnesses and comprise more than 170 types that constantly circulate in humans worldwide. Beyond common colds, rhinoviruses can trigger severe symptoms, particularly in young children, older adults and people with asthma or chronic obstructive pulmonary disease. Despite their clinical and socio-economic impact, no approved vaccine or antiviral treatment exist. Here, we uncovered the interaction of the host AAA+ ATPase RUVBL1/2 with rhinovirus non-structural protein 2C and we demonstrated that RUVBL1/2 is strictly and specifically required for the replication of the viral RNA of the most prevalent and pathogenic rhinovirus species. Pharmacological inhibition of RUVBL1/2 ATPase activity efficiently inhibited rhinovirus replication in a human nasal epithelium model, even post-infection. Moreover, serial viral passaging in the presence of a RUVBL1/2 inhibitor did not lead to the emergence of resistance. These findings reveal an unexpected and strong host dependency with promising potential for antiviral targeting.

microbiology↗

bioRxiv2025

Age-dependent expression and antiviral activity of interferon epsilon in respiratory epithelium

Respiratory syncytial virus (RSV) disease burden is greatest between six weeks and 6 months of life, with young age the most common risk factor among hospitalised children. A robust innate immune response in the airway epithelium is crucial for mitigating RSV-associated disease, but early-life immune responses to infection remain largely unexplored. RNA-seq analysis of RSV-infected primary nasal epithelial cell cultures from healthy infants at birth and one year revealed diminished expression of interferon epsilon (IFNE), a poorly characterised type I IFN, in newborns versus one-year. We hypothesised, therefore, that IFNE plays an important role during infant RSV infection. We found that IFNE is endogenously expressed in airway epithelial cell lines. Recombinant human IFN{varepsilon} (rhIFN{varepsilon}) induced an antiviral state against an RSV clinical isolate and related Sendai virus, but not SARS-CoV-2 under the conditions tested. The antiviral potency of rhIFN{varepsilon} was diminished relative to rhIFN beta (rhIFN{beta}1) or rhIFN lambda-1 (rhIFN{lambda}1), as evidenced by IC50 data. Importantly, rhIFN{varepsilon} induced similar ISGs as rhIFN{beta}1 but demonstrated a transient temporal expression profile that differed from both rhIFN{beta}1 and rhIFN{lambda}1. These results suggest that lower IFN{varepsilon} expression at birth may contribute to increased susceptibility to severe RSV-associated disease, offering insights into potential therapeutic interventions.

microbiology↗

bioRxiv2024

The cold-inducible RNA-binding protein RBM3 stabilises viral mRNA at the cooler temperatures of the upper respiratory tract.

Temperature is a critical determinant of host-pathogen interactions in the respiratory tract, where influenza A virus (IAV) has adapted to the cooler environment of the upper respiratory tract (URT) to enable efficient replication and transmission. The cold-inducible RNA-binding protein RBM3 is highly expressed in nasopharyngeal tissue and is known to stabilise mRNAs under hypothermic conditions; however, its role during viral infection has not been defined. Here, we identify RBM3 as a key host factor that facilitates IAV replication at sub-physiological temperatures. siRNA-mediated knockdown of RBM3 significantly impaired viral replication, while its constitutive overexpression at 37{degrees}C restored replication to levels typically observed at 33{degrees}C. Mechanistically, RBM3 binds directly to viral nucleoprotein (NP) mRNA, prolonging mRNA half-life and inevitably leading to increased viral production. This effect was abolished in an RNA-binding-deficient RBM3 mutant, confirming the requirement for direct RNA interaction, visualised by smiFISH-PLA. Crucially, this positive regulation of NP mRNA and interaction was validated in well-differentiated primary nasal epithelial cells, highlighting the physiological relevance of RBM3 in the human URT. These results reveal a temperature-sensitive host-virus interaction that promotes IAV replication in the cooler URT, a key site for viral shedding and transmission. By linking environmental temperature, host RNA-binding proteins, and viral mRNA stability, this study uncovers a novel mechanism of respiratory virus adaptation and identifies RBM3 as a potential therapeutic target for limiting early-stage viral replication and transmission. Author SummaryTo establish productive infection and ensure transmission, respiratory viruses such as influenza A virus (IAV) must adapt to the diverse environments of the human respiratory tract. One key challenge is the temperature gradient that exists between the upper and lower respiratory tract. The upper respiratory tract (URT), the main site of viral entry and transmission, maintains a lower physiological temperature ([~]33{degrees}C) than the lower respiratory tract ([~]37{degrees}C, LRT), creating a distinct cellular environment. In this study, we investigated how this cooler URT temperature alters the landscape of RNA-binding proteins (RBPs), which play a central role in regulating gene expression after transcription. Using mass spectrometry-based profiling and molecular virology approaches, we identified a cold-inducible RBP, RBM3, as significantly enriched at 33{degrees}C and acting as a key proviral host factor during IAV infection. We show that RBM3 binds directly to viral nucleoprotein (NP) mRNA, mainly in the cytoplasm, in both immortalised cell models and primary nasal epithelial cells. This interaction promotes the stability of NP mRNA thereby enhancing the overall production of infectious virions. Importantly, disrupting RBM3s RNA-binding ability abolished this proviral effect. These findings reveal that RBM3, elevated in the cooler URT environment, directly supports IAV replication. This work highlights how subtle physiological differences in host tissue can reshape post-transcriptional regulation and influence the outcome of respiratory virus infection.

microbiology↗

bioRxiv2021

Inhibition of the TLR4 signalling pathway with TAK-242 reduces RSV infection and cytokine release in primary airway epithelial cells

Respiratory syncytial virus (RSV) infection is the leading cause of hospitalisation in children worldwide, but there is still no vaccine or anti-viral treatment available. RSV has been implicated in the development of respiratory diseases such as asthma. Toll like receptor 4 (TLR4) has been well characterised in the immune responses to RSV. However, the role of TLR4 in RSV infection remains unclear. To study RSV in the lung epithelium, where RSV preferentially infects ciliated cells, we used a well-differentiated primary airway epithelial cell (WD-PAEC) model: a pseudostratified epithelium that produces mucus and beating cilia. We demonstrate in this physiologically relevant model that TLR4 is a pro-viral factor. Inhibition of TLR4 using TAK-242 significantly reduces RSV titres in WD-PAECs in a dose-dependent manner but has no effect on RSV growth kinetics in a range of immortalised respiratory-derived cell lines. Specific inhibition of a range of downstream effectors of TLR4 signalling in the WD-PAEC model identified p38 MAPK as a pro-viral factor, whereas inhibition of MEK1/2 significantly increased RSV titres. Our data demonstrate a role for TLR4 in RSV infection and highlight the importance of biologically relevant models to study virus-host interactions. Author summaryRespiratory Syncytial Virus (RSV) can cause severe respiratory infection in young children and is responsible for approximately 200,000 deaths worldwide every year. Despite decades of research since the identification of this virus in the 1950s there is still no vaccine or treatment available. Advances in research have led to the development of cell cultures that are very similar to the cells that line human airways. These cultures provide an opportunity to study how viruses interacts with airway cells in a representative model and may provide insights that traditional research models have not yet been able to answer. Using this experimental model we show that a drug, TAK-242 which targets a pathogen recognition receptor on the surface of cells, reduces growth of RSV and dampens the immune response to infection in these airway cells. Our data demonstrate potential targets for RSV treatments and also highlight the importance of using relevant experimental models.

microbiology↗

bioRxiv2021

Collaboration Between Host and Viral Factors Shape SARS-CoV-2 Evolution

SARS-CoV-2 continues to evolve, resulting in several variants of concern with novel properties. The factors driving SARS-CoV-2 fitness and evolution in the human respiratory tract remain poorly defined. Here, we provide evidence that both viral and host factors co-operate to shape SARS-CoV-2 genotypic and phenotypic change. Through viral whole-genome sequencing, we explored the evolution of two clinical isolates of SARS-CoV-2 during passage in unmodified Vero-derived cell lines and in parallel, in well-differentiated primary nasal epithelial cell (WD-PNEC) cultures. We identify a consistent, rich genetic diversity arising in vitro, variants of which could rapidly rise to near-fixation with 2 passages. Within isolates, SARS-CoV-2 evolution was dependent on host cells, with Vero-derived cells facilitating more profound genetic changes. However, most mutations were not shared between strains. Furthermore, comparison of both Vero-grown isolates on WD-PNECs disclosed marked growth attenuation mapping to the loss of the polybasic cleavage site (PBCS) in Spike while the strain with mutations in NSP12 (T293I), Spike (P812R) and a truncation of ORF7a remained viable in WD-PNECs. Our work highlights the significant genetic plasticity of SARS-CoV-2 while uncovering an influential role for collaboration between viral and host cell factors in shaping viral evolution and fitness in human respiratory epithelium.

microbiology↗

bioRxiv2020

Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media.

The culture of differentiated human airway epithelial cells allows the study of pathogen-host interactions and innate immune responses in a physiologically relevant in vitro model. As the use of primary cell culture has gained popularity the availability of the reagents needed to generate these cultures has increased. In this study we assessed two different media, Promocell and PneumaCult, during the differentiation and maintenance of well-differentiated primary nasal epithelial cell cultures (WD-PNECs). We compared and contrasted the consequences of these media on WD-PNEC morphological and physiological characteristics and their responses to respiratory syncytial virus (RSV) infection. We found that cultures generated using PneumaCult resulted in greater total numbers of smaller, tightly packed, pseudostratified cells. However, cultures from both media resulted in similar proportions of ciliated and goblet cells. There were no differences in RSV growth kinetics, although more ciliated cells were infected in the PneumaCult cultures. There was also significantly more IL-29/IFN{lambda}1 secreted from PneumaCult compared to Promocell cultures. In conclusion, the type of medium used for the differentiation of primary human airway epithelial cells impacts experimental results.

cell biology↗

bioRxiv2019

Comparative therapeutic potential of ALX-0171 and palivizumab against RSV clinical isolate infection of well-differentiated primary pediatric bronchial epithelial cell cultures

Respiratory syncytial virus (RSV) causes severe lower respiratory tract infections in young infants. There are no RSV-specific treatments available. Ablynx has been developing an anti-RSV F-specific Nanobody(R), ALX-0171. To characterise the therapeutic potential of ALX-0171 we exploited our well-differentiated primary pediatric bronchial epithelial cell (WD-PBEC)/RSV infection model, which replicates several hallmarks of RSV disease in vivo. Using 2 clinical isolates (BT2a; Memphis 37), we compared the therapeutic potential of ALX-0171 with palivizumab, which is currently prescribed for RSV prophylaxis in high-risk infants. ALX-0171 treatment (900 mM) at 24 h post-infection reduced apically released RSV titers to near or below the limit of detection within 24 h for both strains. Progressively lower doses resulted in concomitantly diminished RSV neutralisation. ALX-0171 was approximately 3 fold more potent in this therapeutic RSV/WD-PBEC model than palivizumab (mean IC50 = 346.9-363.6 nM and 1048-1090 nM for ALX-0171 and palivizumab, respectively), irrespective of the clinical isolate. When viral genomic copies (GC) were measured by RT-qPCR, the therapeutic effect was considerably less and GCs were only moderately reduced (0.62 - 1.28 Log10 copies/mL) by ALX-0171 treatment at 300 and 900 nM. Similar findings were evident for palivizumab. Therefore, ALX-0171 was very potent at neutralising RSV released from apical surfaces but only had a limited impact on virus replication. The data indicate a clear disparity between viable virus neutralisation and GC viral load, the latter of which does not discriminate between viable and neutralised RSV. This study validates the RSV/WD-PBEC model for the pre-clinical evaluation of RSV antivirals.

microbiology↗

bioRxiv2018

The Role of Toll-like receptor 4 in respiratory syncytial virus replication, interferon lambda 1 induction, and chemokine responses.

Respiratory syncytial virus (RSV) infection is the leading cause of severe lower respiratory tract infections (LRTI) in infants worldwide. The immune responses to RSV infection are implicated in RSV pathogenesis but RSV immunopathogenesis in humans remains poorly understood. We previously demonstrated that IFN-{lambda}1 is the principle interferon induced following RSV infection of infants and well-differentiated primary pediatric bronchial epithelial cells (WD-PBECs). Interestingly, RSV F interacts with the TLR4/CD14/MD2 complex to initiate secretion of pro-inflammatory cytokines, while TLR4 stimulation with house dust mite induces IFN-{lambda}1 production. However, the role of TLR4 in RSV infection and concomitant IFN-{lambda}1 induction remains unclear. Using our RSV/WD-PBEC infection model, we found that CLI-095 inhibition of TLR4 resulted in significantly reduced viral growth kinetics, and secretion of IFN-{lambda}1 and pro-inflammatory chemokines. To elucidate specific TLR4 signalling intermediates implicated in virus replication and innate immune responses we selected 4 inhibitors, including LY294002, U0126, SB203580 and JSH-23. SB203580, a p38 MAPK inhibitor, reduced both viral growth kinetics and IFN-{lambda}1 secretion, while JSH-23, an NF-{kappa}B inhibitor, reduced IFN-{lambda}1 secretion without affecting virus growth kinetics. Our data indicate that TLR4 plays a role in RSV entry and/or replication and IFN-{lambda}1 induction following RSV infection is mediated, in part, by TLR4 signalling through NF- {kappa}B and/or p38 MAPK. Therefore, targeting TLR4 or downstream effector proteins could present novel treatment strategies against RSV.\n\nImportanceThe role of TLR4 in RSV infection and IFN-{lambda}1 induction is controversial. Using our WD-PBEC model, which replicates many hallmarks of RSV infection in vivo, we demonstrated that the TLR4 pathway is involved in both RSV infection and/or replication and the concomitant induction of IFN-{lambda}1 and other pro-inflammatory cytokines. Increasing our understanding of the role of TLR4 in RSV immunopathogenesis may lead to the development of novel RSV therapeutics.

microbiology↗

bioRxiv2018

The Viral protein Corona Directs Viral Pathogenesis and Amyloid Aggregation

Artificial nanoparticles accumulate a protein corona layer in biological fluids, which significantly influences their bioactivity. As nanosized obligate intracellular parasites, viruses share many biophysical properties with artificial nanoparticles in extracellular environments and here we show that respiratory syncytial virus (RSV) and herpes simplex virus 1 (HSV-1) accumulate a rich and distinctive protein corona in different biological fluids. Moreover, we show that corona pre-coating differentially affects viral infectivity and immune cell activation. Additionally, we demonstrate that viruses bind amyloidogenic peptides in their corona and catalyze amyloid formation via surface-assisted heterogeneous nucleation. Importantly, we show that HSV-1 catalyzes the aggregation of the amyloid beta peptide (A{beta}42), a major constituent of amyloid plaques in Alzheimers disease, in-vitro and in animal models. Our results highlight the viral protein corona as an acquired structural layer that is critical for viral-host interactions and illustrate a mechanistic convergence between viral and amyloid pathologies.

microbiology↗