Biology subjects

Woodall, M. N. J.

Publications and source records attributed to Woodall, M. N. J..

4 recordsLinked to original sources

bioRxiv2026

The canine respiratory epithelium is a permissive ecosystem for influenza interspecies transmission and emergence

The outcome of virus spillover ranges from dead-end infections to pandemics and is underpinned by host-pathogen interactions as well as evolutionary and epidemiological processes. The emergence of novel influenza A viruses (IAVs) has been associated with reassortment events involving multiple species, highlighting the importance of reservoir and intermediate hosts in viral emergence. Highly pathogenic H5N1 IAVs of the 2.3.4.4b genotype have caused a panzootic affecting a broad range of mammals. The role of dogs--arguably the most popular companion animal and a natural host of IAVs--in the ecology of IAVs under this new zooepidemiological scenario is unknown. To address this, we characterised the glycome of the dog respiratory epithelium, infected canine tracheal explants with multiple IAVs (including canine H3N2 and H3N8, equine H3N8, avian H3N8 and H5N1, swine H1N1, human H1N1 and H3N2, and bovine H5N1 viruses), and determined their cellular tropism. We show that the respiratory tract of dogs presents abundant sialylated glycans known to act as IAV receptors. Further, most IAVs (including 2.3.4.4b viruses) infected and replicated in dog tracheas, targeting mainly ciliated cells. Serological testing showed evidence of influenza spillover infections in dogs from the UK. Overall, our results show that the canine respiratory tract can provide a suitable environment for the generation of new IAVs. Given the multi-host contact networks of dogs in nature, they could act as recipients, bridging hosts, and/or mixing vessels for multiple IAV lineages, playing a central role in the ecology of influenza emergence.

microbiology↗

bioRxiv2026

Bovine H5N1 influenza viruses have adapted to more efficiently use receptors abundant in cattle

Sustained mammal-to-mammal transmission of high pathogenicity H5N1 avian influenza viruses is reshaping the host range of these pathogens. One of the longest-running mammalian transmission chains involves the B3.13 genotype circulating in U.S. dairy cattle which was detected in early 2024. Genomic analyses revealed selection and rapid fixation of haemagglutinin mutations D104G and V147M. We demonstate, via glycomic profiling, that bovine tissues, including the mammary gland, are enriched in N- and O-linked glycans capped with N-glycolylneuraminic acid (NeuGc), a sialic acid absent in humans and birds, which instead express only N-acetylneuraminic acid (NeuAc). Early cattle H5 viruses poorly recognized NeuGc, but D104G and V147M enabled efficient engagement of both NeuAc- and NeuGc-containing receptors. These mutations enhanced replication in bovine mammary tissue without major attenuation of replication in human lung and primary nasal epithelial cells. NeuGc-driven receptor adaptation therefore promotes viral fitness in cattle while potentially limiting immediate zoonotic risk. Deep mutational scanning further identified alternative haemagglutinin substitutions that confer NeuGc usage and represent surveillance markers for emerging cattle H5 lineages.

microbiology↗

bioRxiv2023

Lung viral infection modelling in a bioengineered whole-organ

Lung infections are one of the leading causes of death worldwide, and this situation has been exacerbated by the emergence of COVID-19. Pre-clinical modelling of viral infections has relied on cell cultures that lack 3D structure and the context of lung extracellular matrices. Here, we propose a bioreactor-based, whole-organ lung model of viral infection. The bioreactor takes advantage of an automated system to achieve efficient decellularization of a whole rat lung, and recellularization of the scaffold using primary human bronchial cells. Automatization allowed for the dynamic culture of airway epithelial cells in a breathing-mimicking setup that led to an even distribution of lung epithelial cells throughout the distal regions. In the sealed bioreactor system, we demonstrate proof-of-concept for viral infection with the engineered lung by infecting primary human airway epithelial cells. Moreover, to assess the possibility of drug screening in this model, we demonstrate the efficacy of the broad-spectrum antiviral Remdesivir. This whole-organ scale lung infection model represents a step towards modelling viral infection of human cells in a 3D context, providing a powerful tool to investigate the mechanisms of the early stages of pathogenic infections and the development of effective treatment strategies for respiratory diseases.

bioengineering↗

bioRxiv2022

K978C CFTR restores essential epithelial function with greater efficiency than wildtype CFTR when expressed in CF airway cells.

Class Ia/b CFTR variants cause severe cystic fibrosis (CF) lung disease in ~10% of CF patients and are untreatable with small molecule pharmaceuticals. Genetic replacement strategies offer a potential cure for all patients but so far, have displayed limited efficiency in vivo. We hypothesised that increasing protein abundance and/or activity of introduced CFTRs would more effectively restore function to CF bronchial epithelial cells (CFBE) in the presence of CF sputum (CFS) than wildtype (WT)-CFTR. We investigated codon optimised CFTR (hCAI), increased open probability CFTR (K978C) and codon optimised plus K978C (h^K978C) as candidates for gene therapy. Transfection of HEK293T with hCAI and h^K978C produced ~10-fold more CFTR protein than WT or K978C CFTRs. hCAI and h^K978C also displayed ~4-fold greater anion transport than WT in a halide-sensitive YFP quenching assay. However, functionality of modified CFTR cDNAs expressed in CFBE were profoundly different. 10% transduction of CFBE with K978C, compared to 22% transduction with WT, restored Cl- transport to similar levels as that recorded from non-CF cells. K978C increased ASL height and pH more effectively than WT-CFTR, while hCAI and h^K978C had limited impact. Further investigation indicated that codon optimised CFTRs mis-localised in CFBE and compromised vectoral Cl- transport. These data provide further evidence that codon optimised CFTR cDNAs may be unsuitable for gene therapy practices that employ high activity promoters. However, increased activity CFTR cDNAs such as K978C, that potentially mimic the effect of potentiators, may provide more potent recovery of function than WT-CFTR cDNA in CF airways. Significance StatementCystic fibrosis (CF) disease is associated with genetic malfunction of the Cl- channel CFTR, leading to dehydration and decreased pH in the fluid lining the airways. Replacement of CFTR by gene therapy/gene editing offers potential therapeutic benefit but efficiency is poor. We show that gain of activity K978C CFTR under the control of a high activity promoter fully restored Cl- transport, hydration and pH to CF bronchial epithelial cells (CFBE) in the presence of CF sputum and more efficiently than wild type CFTR. Codon optimised forms of CFTR were much less effective and proteins were mis-localised/mis-processed in CFBE. Thus, K978C could offer improved therapeutic potential.

molecular biology↗