Determining age dependent factors driving COVID-19 disease severity using experimental human paediatric and adult models of SARS-CoV-2 infection
Lead Research Organisation:
University College London
Department Name: Institute of Child Health
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a worldwide pandemic of COVID-19. Our lack of understanding about the pathogenesis of SARS-CoV-2 in the human airways is an important barrier to developing effective treatment strategies for COVID-19. So far, we know that COVID-19 disease severity increases with age, with very few severe cases among children. In China, the case fatality ratio was 60-year olds, rising to 15% in those >80.
This project will focus on two aspects of the host response of infection, studying how the cells that line the airway respond to the virus, and whether the recruited immune cells (focussing on the behaviour of neutrophils that are the first to respond to infection) the help reduce viral load or contribute to airway damage and the build-up of debris in the airways. We will use an experimental SARS-CoV-2 infection model of the airway epithelium from young children, adults and the elderly. Our objectives are to determine:
1) if the primary site of infection - the nasal epithelium - of the elderly exhibit increased viral replication and increased inflammatory response to SARS-CoV-2 infection compared to children.
2) the molecular mechanisms that govern age-determinants of COVID-19 disease severity using single-cell genomic analysis of cultured cells and comparing outputs to the same data from age-matched COVID-19 patients using scRNAseq.
3) whether the innate immune response to SARS-CoV2 infected airway epithelium exacerbates inflammation and contributes to the severity of illness in the elderly by measuring a) the intensity of immune cell (neutrophil) recruitment to the airway b) epithelial damage and c) neutrophil phenotype and function.
It is important that we understand the reasons for these fundamental differences in responses in order to help determine what the most appropriate therapy is for COVID-19 disease in these age groups. There is much debate about the utility of novel and exsiting anti-virals as well as immune modulator therapies including NSAIDs and Tocilizumab. This project will rapidly deliver new understanding about the viral pathogenesis and the cause of these age-related disparties in disease outcomes. Not only will this help support the development of effective therapeutics in the short-term, underlining the relevance of this model for the preclinical evaluation of antiviral candidates, but it may also highlight important risk factors or protective mechanisms that could be used to develop early interventional or prophylaxtic therapies in the long-term.
This project will focus on two aspects of the host response of infection, studying how the cells that line the airway respond to the virus, and whether the recruited immune cells (focussing on the behaviour of neutrophils that are the first to respond to infection) the help reduce viral load or contribute to airway damage and the build-up of debris in the airways. We will use an experimental SARS-CoV-2 infection model of the airway epithelium from young children, adults and the elderly. Our objectives are to determine:
1) if the primary site of infection - the nasal epithelium - of the elderly exhibit increased viral replication and increased inflammatory response to SARS-CoV-2 infection compared to children.
2) the molecular mechanisms that govern age-determinants of COVID-19 disease severity using single-cell genomic analysis of cultured cells and comparing outputs to the same data from age-matched COVID-19 patients using scRNAseq.
3) whether the innate immune response to SARS-CoV2 infected airway epithelium exacerbates inflammation and contributes to the severity of illness in the elderly by measuring a) the intensity of immune cell (neutrophil) recruitment to the airway b) epithelial damage and c) neutrophil phenotype and function.
It is important that we understand the reasons for these fundamental differences in responses in order to help determine what the most appropriate therapy is for COVID-19 disease in these age groups. There is much debate about the utility of novel and exsiting anti-virals as well as immune modulator therapies including NSAIDs and Tocilizumab. This project will rapidly deliver new understanding about the viral pathogenesis and the cause of these age-related disparties in disease outcomes. Not only will this help support the development of effective therapeutics in the short-term, underlining the relevance of this model for the preclinical evaluation of antiviral candidates, but it may also highlight important risk factors or protective mechanisms that could be used to develop early interventional or prophylaxtic therapies in the long-term.
Publications
Tommasini F
(2023)
Lung viral infection modelling in a bioengineered whole-organ.
in Biomaterials
Maughan EF
(2022)
Cell-intrinsic differences between human airway epithelial cells from children and adults.
in iScience
Robinson E
(2023)
Trans-epithelial migration is essential for neutrophil activation during RSV infection.
in Journal of leukocyte biology
Ellis S
(2023)
Salivary IgA and vimentin differentiate in vitro SARS-CoV-2 infection: A study of 290 convalescent COVID-19 patients.
in Mucosal immunology
Yoshida M
(2022)
Local and systemic responses to SARS-CoV-2 infection in children and adults.
in Nature
Stephenson E
(2021)
Single-cell multi-omics analysis of the immune response in COVID-19.
in Nature medicine
Woodall M
(2024)
Age-specific nasal epithelial responses to SARS-CoV-2 infection
in Nature Microbiology
Lee DDH
(2022)
COVID-19: Extensive epithelial damage and ciliary dyskinesia in hospitalised patients.
in Rhinology
Kembou-Ringert JE
(2022)
Applications of the hollow-fibre infection model (HFIM) in viral infection studies.
in The Journal of antimicrobial chemotherapy
Woodall MNJ
(2021)
Human models for COVID-19 research.
in The Journal of physiology
Description | When it comes to COVID-19, children are less likely to develop severe respiratory symptoms, while older people are at higher risk of death, even if they have been vaccinated or have access to better treatment options. To better understand why, we have studied how the virus responsible for COVID-19, SARS-CoV-2, affects cells in the nose of people of different ages. Our team of researchers took a multidisciplinary approach to investigate the differences in cellular response to SARS-CoV-2 infection in nasal epithelial cells (NECs) from children, adults, and the elderly. We found that the virus exhibited differential tropism, or the ability to infect different cell types, depending on the age of the host. Ciliated cells are a viral replication center across all age groups, but a distinct goblet inflammatory subtype emerges in infected pediatric cultures, identifiable by high expression of interferon-stimulated genes and truncated viral genomes. Goblet cells are specialized cells that produce mucus to protect the airways, and they play a crucial role in defending against respiratory viruses. We found that infected pediatric goblet cells mounted a robust innate antiviral response to SARS-CoV-2, which correlated with truncated viral reads, greater subgenomic viral RNA, and less infectious progeny compared to older adult cultures. In contrast, infected cultures from elderly individuals showed a proportional increase in ITGB6hi progenitors, which facilitate viral spread and are associated with dysfunctional epithelial repair pathways. NECs are the cells that line the nasal cavity and help to protect against pathogens that enter through the nose. We found that SARS-CoV-2 infection of elderly cultures resulted in epithelial damage and early signs of repair, including cell migration and proliferation of basal NECs to repopulate damaged areas, events that were not detected in cultures derived from younger age groups. Increases in ITGB6, ITGAV and VIM expression, which are associated with epithelial-mesenchymal transition (EMT)and fibrosis, may contribute to the pathological processes seen in elderly people with severe COVID-19. Overall, this study highlights the how the virus affects the airway cells from different age groups and how dysfunctional repair pathways in the elderly may contribute to COVID-19 pathogenesis. |
Exploitation Route | Our findings provide new insights into how the virus affects different age groups and how the nasal epithelium responds to infection. This research will lead to the nucleation of a new research area focused using in vitro models to understand the age-related differences in viral infection and developing targeted therapies for different age groups. The study suggests that dysfunctional repair pathways and the emergence of basaloid-like cells may contribute to COVID-19 pathogenesis in the elderly, which also could potentially open up new avenues for research. The study also highlights the potential role of specific protein, ITGB6, in facilitating viral entry and increasing viral spread in the elderly cells. This finding could lead to new research in understanding how the virus interacts with these proteins and potentially developing drugs that target them to inhibit viral entry. Overall, this study provides important new insights into the pathogenesis of COVID-19 in the elderly and could lead to the nucleation of a new research area focused on understanding the age-related differences in viral infection and developing targeted therapies for different age groups. These insights could aid in the development of new treatments for COVID-19, particularly for older individuals who are at greater risk of severe infection. |
Sectors | Healthcare Pharmaceuticals and Medical Biotechnology |
URL | https://www.biorxiv.org/content/10.1101/2023.01.16.524211v2.full |
Description | Investigating neutrophil function during SARS-CoV2 infection of human airway epithelial cells from different age groups. |
Amount | £86,141 (GBP) |
Funding ID | 182805 |
Organisation | The Child Health Research Charitable Incorporated Organisation |
Sector | Academic/University |
Country | United Kingdom |
Start | 12/2020 |
End | 11/2023 |
Description | Repurposing antiviral drugs for serious viral infections including SARS-CoV-2 |
Amount | £147,667 (GBP) |
Funding ID | V4022 |
Organisation | Great Ormond Street Hospital Children's Charity (GOSHCC) |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 07/2022 |
End | 07/2024 |
Title | COVID-19 across age in vivo integration |
Description | Integration of 8 single-cell datasets comprising 614, 695 cells from upper and lower airways from healthy and COVID-19 patients from paediatric (0-18y), adult (19-50y), and older adult (51-90y) samples. datatsets include: Yoshida, M. et al. Local and systemic responses to SARS-CoV-2 infection in children and adults. Nature vol. 602 321-327 Preprint at https://doi.org/10.1038/s41586-021-04345-x (2022). Loske, J. et al. Pre-activated antiviral innate immunity in the upper airways controls early SARS-CoV-2 infection in children. Nat. Biotechnol. 40, 319-324 (2022). Ziegler, C. G. K. et al. Impaired local intrinsic immunity to SARS-CoV-2 infection in severe COVID-19. Cell 184, 4713-4733.e22 (2021). Melms, J. C. et al. A molecular single-cell lung atlas of lethal COVID-19. Nature 595, 114-119 (2021). Chua, R. L. et al. COVID-19 severity correlates with airway epithelium-immune cell interactions identified by single-cell analysis. Nat. Biotechnol. 38, 970-979 (2020). Trump, S. et al. Hypertension delays viral clearance and exacerbates airway hyperinflammation in patients with COVID-19. Nat. Biotechnol. 39, 705-716 (2021). Delorey, T. M. et al. COVID-19 tissue atlases reveal SARS-CoV-2 pathology and cellular targets. Nature 595, 107-113 (2021). Bharat, A. et al. Lung transplantation for patients with severe COVID-19. Sci. Transl. Med. 12, (2020). |
Type Of Material | Data analysis technique |
Year Produced | 2024 |
Provided To Others? | Yes |
Impact | none yet |
URL | https://www.covid19cellatlas.org/ALI_COVID19/in-vivo/ |
Title | CellxGene Covid19 Cell Atlas for in-vitro ALI dataset |
Description | A web portal to interactively explore or download our RNA-seq data resulting from the in vitro model |
Type Of Material | Data analysis technique |
Year Produced | 2024 |
Provided To Others? | Yes |
Impact | none yet |
URL | https://www.covid19cellatlas.org/ALI_COVID19/in-vitro |
Title | Extended_figures_source_data_images_(Age-specific_nasal_epithelial_responses_to_SARS-CoV-2_infection) |
Description | Extended_figures_source_data_images_for manuscript: Age-specific_nasal_epithelial_responses_to_SARS-CoV-2_infection |
Type Of Material | Database/Collection of data |
Year Produced | 2024 |
Provided To Others? | Yes |
Impact | none yet |
URL | https://figshare.com/articles/media/Extended_figures_source_data_images_Age-specific_nasal_epithelia... |
Title | Main_figures_source_data_images_(Age-specific_nasal_epithelial_responses_to_SARS-CoV-2_infection) |
Description | Main_figures_source_data_images_for manuscript: Age-specific_nasal_epithelial_responses_to_SARS-CoV-2_infection |
Type Of Material | Database/Collection of data |
Year Produced | 2024 |
Provided To Others? | Yes |
Impact | none yet |
URL | https://figshare.com/articles/media/Main_figures_source_data_images_Age-specific_nasal_epithelial_re... |
Title | Supplementary_files_source_data_images_(Age-specific_nasal_epithelial_responses_to_SARS-CoV-2_infection) |
Description | Supplementary material 1: Whole mount stain of a 72hr infected Nasal epithelial culture with wounding. Images show stains for DAPI, dsRNA and Phalloidin. Image shows spread of dsRNA and increased signal at the wound site.Supplementary material 2: Transmission electron micrographs of Paediatric Goblet 2 inflammatory cells. Transmission electron micrographs of Goblet cells from a paediatric donor 72h p.i. with SARS-CoV-2. Master panels show different magnifications, the scale bar is given on the left (top panels) or above (bottom panels) of each image. Viral particles are indicated with white arrows.Supplementary material 3: Transmission electron micrograph of epithelial cell damage at 72h p.i with SARS-CoV-2. Master panel displays virions (white arrows), viral compartments (VC), Cytopathology including endocytosed cilia basal bodies, loss of tight junctions (grey arrows). |
Type Of Material | Database/Collection of data |
Year Produced | 2024 |
Provided To Others? | Yes |
Impact | none yet |
URL | https://figshare.com/articles/media/Supplementary_files_source_data_images_Age-specific_nasal_epithe... |
Description | UCL-Sanger collaboration |
Organisation | The Wellcome Trust Sanger Institute |
Country | United Kingdom |
Sector | Charity/Non Profit |
PI Contribution | Samples provided to Sanger |
Collaborator Contribution | scRNAseq of human airway epithelail cells infected with SARS-CoV-2 or controls. Bioinformatics analysis |
Impact | None yet |
Start Year | 2021 |
Description | Image competition |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | A magazine article about the Great Ormond Street Hospital for Children NHS Foundation Trust research image competition 'A moment of Research' - Our entry, an image of adult nasal airway cells in culture was taken by our research technician Katie-Marie Case. Her image was on the shortlist of winners and was entitled 'From the Milky way to the Airway' |
Year(s) Of Engagement Activity | 2022 |
URL | https://www.sciencefocus.com/news/kaleidoscope-great-ormond-streets-image-awards/ |
Description | Lab Website |
Form Of Engagement Activity | Engagement focused website, blog or social media channel |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | In Feb 2022 I developed a website using MS Sway to showcase our research with links to open-accesss publications. MS Sway provides viewer analytics and so far this has been viewed 9 times. It is only accessible through a link on the UCL IRIS system for the PI at the moment, but this will continue to be developed upon and advertised widely using social media and UCL channels. |
Year(s) Of Engagement Activity | 2022 |
URL | https://sway.office.com/T9xNUhKrV0RgmvUe |
Description | Modelling COVID-19 with Air-Liquid Interface Cultures (for British Asscoaition for Lung Research (BALR) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Other audiences |
Results and Impact | presntation of our research models for clinicians, academics and researchers in lung research field |
Year(s) Of Engagement Activity | 2020 |
URL | https://www.balr.co.uk/meetings |
Description | Nature: The best science images of 2022 |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | Nature shortlisted 17 images as the Best Scince Image of 2022 and an image catured duing this study was included. This image was entitled 'Nasal swirl' with the caption:. These human nasal cells are covered in cilia - tiny hairs that trap and clear foreign bodies from the nose. While studying why COVID-19 affects certain age groups more than others, PhD student Katie-Marie Case noticed that these galaxy-like nasal cell spirals were present only in older patients. |
Year(s) Of Engagement Activity | 2022 |
URL | https://www.nature.com/immersive/d41586-022-04372-2/index.html |
Description | Observing the Airway Barrier during SARS-CoV2 Infection. For Physiological Society |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | The conference reviewed the challenges of understanding the pathophysiological changes occurring throughout the body following COVID-19 infection. The conference also shared current knowledge and thinking across many physiological systems, showcased the symbiotic relationship between physiology and clinical care and helped set the agenda for research to identify future treatments and therapies. |
Year(s) Of Engagement Activity | 2020 |
URL | https://www.youtube.com/watch?v=v2Y4uP0aiV8 |
Description | Science Focus: The moment of discovery: 11 massive medical breakthroughs, in pictures |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Another image captured duirng this study, this time using electron microscopy, was shortlisted as one of 11 as part of an artuicle in BBC Sceince Focus magazine. |
Year(s) Of Engagement Activity | 2023 |
URL | https://www.sciencefocus.com/the-human-body/the-moment-of-discovery-11-massive-medical-breakthroughs... |
Description | Video Abstract of Models used in this study on YouTube |
Form Of Engagement Activity | Engagement focused website, blog or social media channel |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Video abstract of a review article describing the types of methods that can be used to study COVID19 disease |
Year(s) Of Engagement Activity | 2021 |
URL | https://www.youtube.com/watch?v=DotawqMABDY&t=11s |
Description | Video Abstract of Work |
Form Of Engagement Activity | Engagement focused website, blog or social media channel |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | A video abtract was made by the PI and involved the first author of the paper to showcase the main findings of the work. This led to training in video content for the postdoc and outreach for our work. |
Year(s) Of Engagement Activity | 2022,2023 |
URL | https://youtu.be/uExP4bx6D_A |