Investigating the role of ANP32A in the replication of Avian influenza Virus
Lead Research Organisation:
Imperial College London
Department Name: Infectious Disease
Abstract
Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
Technical Summary
Influenza viruses infect domestic animals including poultry and pigs, causing loss of income, social and economic disruption. Avian influenza remains one of the major threats to food security. Adaptation of avian influenza in animal hosts leads to viruses that are transmitted in humans and carries the risk of driving emergent influenza pandemics. We have investigated a family of host proteins, ANP32, that are co-opted by influenza virus to support its replication. The difference between ANP32A in chickens and mammalian hosts accounts for one of the host range barriers that protect pigs and humans from zoonotic infections by avian influenza viruses. Avian influenza virus RNA-dependent RNA polymerase is unable to utilise the mammalian ANP32A protein unless it adapts by mutation of the viral PB2 protein. We will first identify the specific regions of ANP32A needed for viral protein interactions. We will use new genome editing tools that we have developed to modify chicken cells, to identify functional regions/amino acids by testing the effectiveness of specific mutations to abrogate these host:viral interactions. We will determine any global changes in the RNA transcriptome of the edited cells containing modified ANP32A protein and characterise any other effects of these modifications, unrelated to avian influenza infection. We will investigate the function of ANP32A in genome edited animals.
Planned Impact
Agricultural losses to avian influenza have major impacts through recurring outbreaks in tropical environments mostly in developing countries. Small holder farmers and subsistence farmers lose valuable resources (in terms of livestock) during these outbreaks. The Gates Foundation has invested in promoting scavenger chicken to improve the livelihoods of subsistence farmers, with a strong focus on women who are usually the primary owners of rural chicken. The Gates Foundation is also investing in research at The Roslin Institute (through the Centre for Tropical Livestock Genetics and Health) to develop disease resistance using genetic approaches in poultry, to alleviate the need for vaccination programmes.
Agricultural losses by high pathogenic avian influenza are also severe in developed countries. The 'bird flu' outbreak in the USA in 2015 led to the culling of 48 million layer chickens and turkeys. The economic loss from this outbreak is estimated to be $1.3 billion. Most of these losses were due to the loss of the export market as live poultry products can no longer be exported from affected countries during HPAI incidents.
The major advances in understanding the interaction between specific host genes and influenza virus replication that we propose to investigate here will inform genetic strategies for control of avian influenza in production chickens.
Another primary beneficiary of our research will be the research community, especially those researchers working in avian biology or poultry production. We will exemplify the development and use of new genetic tools for investigating gene function and viral resistance in poultry. This will enable researchers to replace some animal model challenges with cell-based challenges. Poultry researchers will also benefit by the example of creating chicken genetic models that can be used to investigate pathways of infections and resistance.
Agricultural losses by high pathogenic avian influenza are also severe in developed countries. The 'bird flu' outbreak in the USA in 2015 led to the culling of 48 million layer chickens and turkeys. The economic loss from this outbreak is estimated to be $1.3 billion. Most of these losses were due to the loss of the export market as live poultry products can no longer be exported from affected countries during HPAI incidents.
The major advances in understanding the interaction between specific host genes and influenza virus replication that we propose to investigate here will inform genetic strategies for control of avian influenza in production chickens.
Another primary beneficiary of our research will be the research community, especially those researchers working in avian biology or poultry production. We will exemplify the development and use of new genetic tools for investigating gene function and viral resistance in poultry. This will enable researchers to replace some animal model challenges with cell-based challenges. Poultry researchers will also benefit by the example of creating chicken genetic models that can be used to investigate pathways of infections and resistance.
Publications

Carrique L
(2020)
Host ANP32A mediates the assembly of the influenza virus replicase.
in Nature

Idoko-Akoh A
(2023)
Creating resistance to avian influenza infection through genome editing of the ANP32 gene family.
in Nature communications


Long JS
(2019)
Species specific differences in use of ANP32 proteins by influenza A virus.
in eLife

McKay PF
(2022)
Polymer formulated self-amplifying RNA vaccine is partially protective against influenza virus infection in ferrets.
in Oxford open immunology

Mistry B
(2020)
Elucidating the Interactions between Influenza Virus Polymerase and Host Factor ANP32A.
in Journal of virology

Peacock T
(2020)
Swine ANP32A supports avian influenza virus polymerase


Peacock TP
(2021)
SARS-CoV-2 one year on: evidence for ongoing viral adaptation.
in The Journal of general virology

Peacock TP
(2020)
Swine ANP32A Supports Avian Influenza Virus Polymerase.
in Journal of virology
Description | Work funded by this award has identified specific mutations which render the chicken host factor ANP32A unable to support replication of flu. Mutations at positions 129 and 130 of the chicken ANP32A gene have been introduced into chicken germ cells to demonstrate loss of support of flu virus in these cells. However, some replication can still occur so we are now looking to characterise escape mutants and to use recent structural data to identify specific mutations at the N-terminus of ANP32A in order to abrogate its recruitment of a second polymerase. Work is also ongoing into the potential role of ANP32E in support of influenza polymerase since it might be that editing of this second chicken gene will be require to completely abrogate influenza infection. We have now identified several different escape mutations in the viruses isolated from gene edited chickens that enable the virus to continue to replicate using the modified host factor. We can see that these affect the dimerization potential of the polymerase enzyme and this is revealing key regulatory mechanisms. we can also see the mutations may make it more likely that the virus can replicate in mammalian cells using mammalian ANP32 proteins that are shorted than avian ANP32A. This is worrisome and make the impact of our finding very significant. We are preparing a high profile publication to share these findings widely. |
Exploitation Route | These findings can be applied directly to the production of gene edited chickens which are resistant to flu. This has the potential to reduce bird culling in the poultry industry in response to flu outbreaks and potentially to reduce the incidence of human flu pandemics by removing poultry as a source of novel reassortant viruses. However importantly our findings show that simple modifications may also lead to virus mutations and the design of gene edits must be carefully assessed. |
Sectors | Agriculture Food and Drink |
Description | A patent application is being considered, based on our findings, to generate influenza resistant chickens. Advice has been given to policy makers around the generation and use of gene edited animals for farming. |
First Year Of Impact | 2022 |
Sector | Agriculture, Food and Drink |
Impact Types | Policy & public services |
Description | Nervtag advisor |
Geographic Reach | National |
Policy Influence Type | Implementation circular/rapid advice/letter to e.g. Ministry of Health |
Impact | Nervtag Advisor |
Description | invited talk to USA National academy of Science around genome edited animals |
Geographic Reach | North America |
Policy Influence Type | Participation in a guidance/advisory committee |
Description | Roslin collaboration |
Organisation | University of Edinburgh |
Department | The Roslin Institute |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We have collaborated with this group and are publishing a paper together, we have shared pertinent unpublished data together as well as laboratory reagents |
Collaborator Contribution | We have collaborated with this group and are publishing a paper together, we have shared pertinent unpublished data together as well as laboratory reagents |
Impact | A paper is currently in revisions and should be published soon in Journal of Virology. |
Start Year | 2019 |
Description | 11th October 2023 Creating resistance to avian influenza infection by genome editing if the anp32 gene family interview for the Today Programme BBC radio interviewed by elena.angelides.ext@bbc.co.uk; |
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 | Title Creating resistance to avian influenza infection by genome editing if the anp32 gene family |
Year(s) Of Engagement Activity | 2024 |
Description | Astbury conversation "Inside the cell with influenza virus." |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | Talk |
Year(s) Of Engagement Activity | 2022 |
URL | https://astburyconversation.leeds.ac.uk/ehome/index.php?eventid=200183132& |
Description | Australasian Virology Society's 21st Anniversary Meeting (AVS11), held in Australia, 5-8 December 2022. |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Virology in Australia to share and encourage collaborations and exchange of knowledge |
Year(s) Of Engagement Activity | 2022 |
URL | https://www.avs.org.au/ |
Description | BBC 1 radio inside science 7th February 2024 |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | general news item on BBC radio |
Year(s) Of Engagement Activity | 2024 |
Description | Cardiff School of Medicine, regular Science Seminar Series, Wendy Barclay talk title Perspectives on Pandemics |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | . The seminar is circulated across several Schools and posted on Eventbrite. Talks are 45-50 minutes plus questions and the audience is multi-disciplinary - immunologists, bioinformaticians, modellers etc |
Year(s) Of Engagement Activity | 2023 |
Description | Cardiff University Infection & Immunity meeting 2022 'Saving the world from the next pandemic' |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | Infection and Immunity annual meeting 2022 Cardiff City Stadium, Leckwith Road, Cardiff, CF11 8AZ, United Kingdom |
Year(s) Of Engagement Activity | 2022 |
URL | https://www.cardiff.ac.uk/systems-immunity |
Description | Clare Wilson |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | Interview about an article about bird flu - in a Nature paper finding that a gene in humans is important in determining human resistance to avian flu viruses, and identified mutations in virus genes that let them overcome this resistance. The authors have just held a press conference where they said it will be important to check for such mutations in pandemic flu surveillance processes. |
Year(s) Of Engagement Activity | 2023 |
Description | DMM Meeting Wendy Barclay, Organiser, Chair |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Disease Models & Mechanisms Meeting Infectious Diseases Through an Evolutionary Lens London, UK, Tuesday 17 - Thursday 19 October 2023 |
Year(s) Of Engagement Activity | 2023 |
URL | https://www.biologists.com/meetings/dmminfectious2023/ |
Description | Microbiology Seminar Series, washington University in St Louis USA. Wendy Barclay Keynote speaker Seminar Title: ANP:32 a key host cell factor for influenza virus replication |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Microbiology Seminar Series, Washington University in St Louis School of Medicine. |
Year(s) Of Engagement Activity | 2023 |
Description | NSV portugal organiser XVIII International Conference on Negative Strand Viruses |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | co-organiser |
Year(s) Of Engagement Activity | 2022 |
Description | Ri.Med Symposium Palermo, Italy October 23 Wendy Barclay keynote speaker ANP32: A key host factor for influenza virus replication , |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | lecture ANP32: A key host factor for influenza virus replication Wendy Barclay, |
Year(s) Of Engagement Activity | 2023 |
Description | Wendy Barclay invited speaker at the 70th Annual meeting of the Japanese Society for Virology, in Sendai, Japan September 2023 |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Keynote speaker - title of talk Innovation in vaccine development: Perspectives from global research leaders. |
Year(s) Of Engagement Activity | 2023 |
Description | isirv - OPTIONS for the Control of Influenza XI Belfast Keynote speaker talk title Influenza research in the post COVID era |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | To share knowledge and further study Influenza |
Year(s) Of Engagement Activity | 2022 |
URL | https://www.isirv.org/site/ |
Description | royal society : UK-Switzerland BIM June 2023 |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Wendy Barclay keynote talk: Host barriers to the emergence of influenza pandemics |
Year(s) Of Engagement Activity | 2023 |