The role of LC3-associated phagocytosis during virus infection
Lead Research Organisation:
University of Liverpool
Department Name: Institute of Infection and Global Health
Abstract
LC3-associated phagocytosis (LAP) is a recently-characterised phagocytic pathway that plays important roles during immunity to infectious agents including viruses. LAP shares many features with the well-characterised autophagy pathway but differs in signalling events upstream of LC3 recruitment to membranes. Thus, the precise role played by each pathway 'in vivo' has been very difficult to dissect. The unique aspect of this grant proposal is based on our generation of mice that are deficient in LAP, but have normal autophagy. This allows us to focus specifically on the role played by LAP in the control of viral infection. Our preliminary data using Influenza A virus (IAV) infection of LAP deficient mice reveal that there are higher virus titres, more inflammation and more clinical signs in the absence of LAP. We therefore believe that LAP plays an important role in host defence in the respiratory tract.
The aims of this project are to:
1. Discover the precise way that LAP acts in the defence mechanisms to IAV infection. This will involve comparing the progress of viral infection in genetically modified mice deficient in LAP with normal mice. We will also study whether LAP influences inflammatory responses, various types of immune responses (antibody and cytotoxic T cell) as well as other anti-viral responses such as interferon.
2. Discover whether LAP specifically in macrophages or epithelial cells plays a role in the defence against IAV. To do this, we generate conditional knockout mice that are lacking LAP in macrophages but not in all other tissues. These mice will be infected with IAV and then investigated as in Aim 1.
The results will significantly enhance our understanding of fundamental aspects of defence to virus infection as well as aspects of respiratory biology.
The research will be carried out at the Universities of Liverpool and UEA by a multi-disciplinary team comprising members of the Medical and Veterinary Faculties using well-equipped facilities currently situated at these sites. Defects in autophagy are associated with aberrant host defence, inflammatory disease and age-related disorders. It is possible that a defect in LAP, rather than autophagy, accounts for some of these pathological conditions. Characterisation LAP separate from autophagy, will allow examination of pathologies linked to autophagic machinery in a new light. In turn this may lead to novel approaches to improving antiviral therapies or therapeutic interventions against the consequences of viral infection
The aims of this project are to:
1. Discover the precise way that LAP acts in the defence mechanisms to IAV infection. This will involve comparing the progress of viral infection in genetically modified mice deficient in LAP with normal mice. We will also study whether LAP influences inflammatory responses, various types of immune responses (antibody and cytotoxic T cell) as well as other anti-viral responses such as interferon.
2. Discover whether LAP specifically in macrophages or epithelial cells plays a role in the defence against IAV. To do this, we generate conditional knockout mice that are lacking LAP in macrophages but not in all other tissues. These mice will be infected with IAV and then investigated as in Aim 1.
The results will significantly enhance our understanding of fundamental aspects of defence to virus infection as well as aspects of respiratory biology.
The research will be carried out at the Universities of Liverpool and UEA by a multi-disciplinary team comprising members of the Medical and Veterinary Faculties using well-equipped facilities currently situated at these sites. Defects in autophagy are associated with aberrant host defence, inflammatory disease and age-related disorders. It is possible that a defect in LAP, rather than autophagy, accounts for some of these pathological conditions. Characterisation LAP separate from autophagy, will allow examination of pathologies linked to autophagic machinery in a new light. In turn this may lead to novel approaches to improving antiviral therapies or therapeutic interventions against the consequences of viral infection
Technical Summary
LC3-associated phagocytosis (LAP) is a recently characterised phagocytic pathway that plays important roles during innate defence and immunity to pathogens. LAP shares many components with the well-characterised autophagy pathway but differs in signalling events upstream of LC3 recruitment to membranes. Thus, the precise role played by each pathway 'in vivo' has been very difficult to dissect. The unique aspect of this grant proposal is based on our generation of mice that are deficient in LAP, but have normal autophagy. This allows us to focus specifically on the role played by LAP in the control of viral infection. Our preliminary data using Influenza A virus (IAV) infection of LAP deficient mice reveal that there are higher virus titres, more inflammation and more clinical signs in the absence of LAP. We therefore believe that LAP plays an important role in host defence in the respiratory tract.
We will use mice deficient in LAP, and mice where LAP is specifically removed from macrophages or epithelial cells. The transgenic models will be infected with IAV and the pathogenesis of infection analysed for virological, inflammatory and immunological parameters. IAV is relevant to disease in animals and has good sets of specific reagents that will allow the study of the host response in mice. This will allow us to determine the role played by LAP during an acute, cytopathic infection in the respiratory tract that is normally cleared rapidly generating immunity to re-infection.
The results will significantly enhance our understanding of fundamental aspects of host responses to virus infection in the lung as well as respiratory biology. In turn this may lead to novel approaches to improving antiviral therapies or therapeutic interventions against the consequences of viral infection.
We will use mice deficient in LAP, and mice where LAP is specifically removed from macrophages or epithelial cells. The transgenic models will be infected with IAV and the pathogenesis of infection analysed for virological, inflammatory and immunological parameters. IAV is relevant to disease in animals and has good sets of specific reagents that will allow the study of the host response in mice. This will allow us to determine the role played by LAP during an acute, cytopathic infection in the respiratory tract that is normally cleared rapidly generating immunity to re-infection.
The results will significantly enhance our understanding of fundamental aspects of host responses to virus infection in the lung as well as respiratory biology. In turn this may lead to novel approaches to improving antiviral therapies or therapeutic interventions against the consequences of viral infection.
Planned Impact
Respiratory virus infections are of global significance to both the human and animal populations. The zoonotic potential of influenza and the implications of emerging new strains are well recognised by both the scientific community and the general public. Improved understanding of the pathogenesis of infection and the host response to these pathogens is critical in improving treatment and management of respiratory disease and the associated morbidity and mortality. This work aims to further this understanding by looking at the role of LC3-associate phagocytosis (LAP) in the innate and adaptive responses of the host respiratory tract following infection with influenza virus.
The academic impact of this work therefore will be to further the knowledge of the host response to viral infection, providing the foundations of academic knowledge and understanding on which future advancements in treatment and disease control can be built. The use of the multi-disciplinary approach as proposed here brings together expertise in cell biology, molecular virology, immunology and veterinary pathology, thereby maximising the potential outputs of the research. This is also advantageous to the RA in providing the experience and training involving multiple fields fostering a multi-disciplinary approach and its advantages for enhanced, productive science as well as allowing them to develop a range of transferable skills that will enhance their career development. This is achieved by collaboration between the Universities of East Anglia and Liverpool.
The societal impact of furthering the understanding of the host response to infection with Influenza virus on the health of the individual and the wider population is significant. Contribution to the improved treatment of individual high risk patients where infection with Influenza virus has a higher morbidity and mortality than the general population, or increased understanding of the risk factors associated with epidemic strains, both are important in the future of prevention and control of Influenza infection, in both veterinary species and the human population.
The academic impact of this work therefore will be to further the knowledge of the host response to viral infection, providing the foundations of academic knowledge and understanding on which future advancements in treatment and disease control can be built. The use of the multi-disciplinary approach as proposed here brings together expertise in cell biology, molecular virology, immunology and veterinary pathology, thereby maximising the potential outputs of the research. This is also advantageous to the RA in providing the experience and training involving multiple fields fostering a multi-disciplinary approach and its advantages for enhanced, productive science as well as allowing them to develop a range of transferable skills that will enhance their career development. This is achieved by collaboration between the Universities of East Anglia and Liverpool.
The societal impact of furthering the understanding of the host response to infection with Influenza virus on the health of the individual and the wider population is significant. Contribution to the improved treatment of individual high risk patients where infection with Influenza virus has a higher morbidity and mortality than the general population, or increased understanding of the risk factors associated with epidemic strains, both are important in the future of prevention and control of Influenza infection, in both veterinary species and the human population.
Publications
Box HJ
(2024)
Lack of antiviral activity of probenecid in vitro and in Syrian golden hamsters.
in The Journal of antimicrobial chemotherapy
Charman M
(2021)
Constitutive TRIM22 Expression in the Respiratory Tract Confers a Pre-Existing Defence Against Influenza A Virus Infection.
in Frontiers in cellular and infection microbiology
Gallardo-Toledo E
(2023)
Chemoprophylactic Assessment of Combined Intranasal SARS-CoV-2 Polymerase and Exonuclease Inhibition in Syrian Golden Hamsters
in Viruses
Heckmann B
(2020)
Noncanonical function of an autophagy protein prevents spontaneous Alzheimer's disease
in Science Advances
Hiscox JA
(2021)
Shutting the gate before the horse has bolted: is it time for a conversation about SARS-CoV-2 and antiviral drug resistance?
in The Journal of antimicrobial chemotherapy
Neary M
(2023)
Evaluation of Nafamostat as Chemoprophylaxis for SARS-CoV-2 Infection in Hamsters.
in Viruses
Serramito-Gómez I
(2020)
Regulation of cytokine signaling through direct interaction between cytokine receptors and the ATG16L1 WD40 domain.
in Nature communications
Wang Y
(2022)
Control of infection by LC3-associated phagocytosis, CASM, and detection of raised vacuolar pH by the V-ATPase-ATG16L1 axis.
in Science advances
Wang Y
(2021)
Non-canonical autophagy functions of ATG16L1 in epithelial cells limit lethal infection by influenza A virus.
in The EMBO journal
Description | Non-canonical autophagy is a cellular process that this conventionally thought of as being involved in the function of phagocytic cells and how they are able to mediate defence against invading pathogens. We have shown that during influenza infection non-canonical autophagy has an important role to play in the pathogenesis of influenza disease and that this process is important in the epithelial barrier not phagocytic cells. We predicted that cells involved in generating immune responses like anti-viral antibodies (dentritic cells and macrophages) would be affected by a lack of non-canonical autophagy. However, we have found that these so-called antigen presenting functions are not affected by non-canonical autophagy. |
Exploitation Route | Modulation of non-canonical autophagy by drugs might be a means of improving outcomes in influenza (and other respiratory virus such as SARS-CoV-2) infections |
Sectors | Agriculture Food and Drink Pharmaceuticals and Medical Biotechnology |
Description | Digard |
Organisation | University of Edinburgh |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Infection model of 3D tracheal epithelial cell cultures derived from wild type and BPIFA1 KO mice |
Collaborator Contribution | Reagents and expertise in terms of labelled IAV and highly concentrated preps of IAV |
Impact | Determination that BPIFA1 is involved in binding and entry of IAV into epithelial cells |
Start Year | 2015 |
Description | SPLUNC1 Tripp |
Organisation | University of Georgia |
Country | United States |
Sector | Academic/University |
PI Contribution | The Stewart group are investigating the host response to virus infection in mouse models |
Collaborator Contribution | The Tripp group have supplied expertise and access to reagents such as influenza strains as well a performed infections in mice with highly pathogenic influenza strains. |
Impact | 1. Leeming, G.H., Kipar, A., Hughes, D.J., Bingle, L., Bennett, E., Moyo, N., Tripp, R.A., Bigley, A., Bingle, C.D., Sample, J.T., Stewart, J.P. Gammaherpesvirus infection modulates the temporal and spatial expression of SCGB1A1 (CCSP) and BPIFA1 (SPLUNC1) in the respiratory tract. Laboratory Investigation, In Press. |
Start Year | 2011 |
Description | Simon Carding vaccine |
Organisation | Quadram Institute Bioscience |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Design and preliminary testing of bacterial OMVs containing influenza proteins for use as a mucosal vaccine against avian influenza |
Collaborator Contribution | Construction and production of bacterial OMVs containing influenza proteins for use as a mucosal vaccine against avian influenz |
Impact | none as yet |
Start Year | 2018 |
Description | Interview for Mail on Sunday |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Interview with Mail on Sunday journalist regarding co0infection with influenza A and in particular the influence of COVID on infleunza case rates |
Year(s) Of Engagement Activity | 2020 |
URL | https://www.dailymail.co.uk/health/article-8875201/Has-Covid-killed-flu.html |
Description | Interview for Mail on Sunday |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Interview with MAil on Sunday talking about the case rates for influenza for a second year and how they had been influenced by the COVID pandemic. Also regarding hte case rates of avian influenza and the risk to human health. |
Year(s) Of Engagement Activity | 2021 |
URL | https://www.dailymail.co.uk/health/article-10324309/Covids-killed-flu-SECOND-year.html |
Description | Podcast interview |
Form Of Engagement Activity | A broadcast e.g. TV/radio/film/podcast (other than news/press) |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Media (as a channel to the public) |
Results and Impact | Podacast for Guardian Science Weekly about how respiratory viruses cause disease focussed around the Covid-19 epidemic. |
Year(s) Of Engagement Activity | 2020 |