chIFITM knockdown/knockout technology as a platform technology for increased vaccine yields.

Lead Research Organisation: The Pirbright Institute
Department Name: Pirbright Laboratory

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.

Publications

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Description This Funding provided essential data for a successful application to the Bill and Melinda Gates foundation.
Exploitation Route Numerous vaccines are produced in hen's eggs or cell lines, but the amount that can be produced is limited by the immune responses that prevent replication of vaccine viruses. The funding will enable previous research carried out by the Genetics and Genomics group at Pirbright to be continued. The researchers were the first to describe a set of immune proteins in the chicken called chIFITMs, which prevent viruses from multiplying in cells.

Many vaccines, for both animal and human, are produced by growing a weakened form of the virus in chicken eggs or cells, which are then extracted for use. Although chIFITM may help protect chickens from viral infection, the protein actually hinders vaccine production, as it prevents the weakened virus from replicating at high levels and reduces the amount of vaccine that can be made.

Our new research will involve using a gene editing system called CRISPR/Cas9 to remove the chIFITM genes in chicken cells, therefore overcoming one of the barriers for viral replication, and boosting the levels of vaccine virus produced. For example, the flu vaccine currently requires two eggs to produce a single dose, but inactivating chIFITM genes could mean only a single egg is needed per dose.

This increase in yield will make vaccines cheaper to produce and more accessible to livestock owners in developing nations, the main target of this funding. Initially the scientists will focus on increasing flu vaccine yields, but the technique can be applied to multiple livestock viral diseases and potentially human diseases too. Once the researchers have piloted this technique, for which a patent has been filed, they will work with commercial partners to bring this new technology to market.

By 2024, global poultry meat consumption is expected to rise by over 20 million tonnes compared to 2015, and it is therefore essential to maximise outputs worldwide. Poultry production is persistently reduced through viral infection; developing efficient and affordable vaccines against viral diseases will alleviate poverty in developing countries, where these diseases cause devastating consequences for subsistence poultry farming.
This research has the potential to increase vaccine yields between five and ten-fold, which, in an industry where the total revenue for vaccines produced in eggs and cell lines is approximately $14.3 billion, can make vast differences to manufacturing costs which will in turn enable vaccine prices to be reduced. The project is set to run for four years, so this new technology could be commercially available as early as 2021.
Sectors Agriculture, Food and Drink,Healthcare,Pharmaceuticals and Medical Biotechnology

URL https://www.pirbright.ac.uk/our-science/avian-viral-diseases/genetics-and-genomics
 
Description Numerous vaccines are produced in hen's eggs or cell lines, but the amount that can be produced is limited by the immune responses that prevent replication of vaccine viruses. The funding will enable previous research carried out by the Genetics and Genomics group at Pirbright to be continued. The researchers were the first to describe a set of immune proteins in the chicken called chIFITMs, which prevent viruses from multiplying in cells. Many vaccines, for both animal and human, are produced by growing a weakened form of the virus in chicken eggs or cells, which are then extracted for use. Although chIFITM may help protect chickens from viral infection, the protein actually hinders vaccine production, as it prevents the weakened virus from replicating at high levels and reduces the amount of vaccine that can be made. Our new research will involve using a gene editing system called CRISPR/Cas9 to remove the chIFITM genes in chicken cells, therefore overcoming one of the barriers for viral replication, and boosting the levels of vaccine virus produced. For example, the flu vaccine currently requires two eggs to produce a single dose, but inactivating chIFITM genes could mean only a single egg is needed per dose. This increase in yield will make vaccines cheaper to produce and more accessible to livestock owners in developing nations, the main target of this funding. Initially the scientists will focus on increasing flu vaccine yields, but the technique can be applied to multiple livestock viral diseases and potentially human diseases too. Once the researchers have piloted this technique, for which a patent has been filed, they will work with commercial partners to bring this new technology to market.
First Year Of Impact 2017
Sector Agriculture, Food and Drink,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
Impact Types Societal

 
Description LVIF
Amount $1,500,000 (CAD)
Organisation Bill and Melinda Gates Foundation 
Sector Charity/Non Profit
Country United States
Start 04/2018 
End 09/2020
 
Description Pathfinder BB/R012431/1 Grant holder: Dr Mark Fife. Grant title: chIFITM knockdown/knockout technology as a platform technology for increased vaccine yields in SPF Eggs.
Amount £12,500 (GBP)
Funding ID BB/R012431/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 11/2017 
End 02/2018
 
Title Comprehensive analysis of genetic variants across immune loci in indigenous breeds. 
Description The interferon-induced transmembrane (IFITM) protein family comprises a class of restriction factors widely characterised in humans for their potent antiviral activity. Their biological activity is well documented in several animal species, but their genetic variation and biological mechanism is less well understood, particularly in avian species. Here we report the complete sequence of the domestic chicken Gallus gallus IFITM locus from a wide variety of chicken breeds to examine the detailed pattern of genetic variation of the locus on chromosome 5, including the flanking genes ATHL1 and B4GALNT4. We have generated chIFITM sequences from commercial breeds (supermarket-derived chicken breasts), ancient bone samples, indigenous chickens from Nigeria (Nsukka) and Ethiopia, European breeds and inbred chicken lines from The Pirbright Institute, totalling of 211 chickens. Through mapping of genetic variants to the latest chIFITM consensus sequence our data reveal that the chIFITM locus does not show structural variation in the locus across the populations analysed, despite spanning diverse breeds from different geographic locations. However, single nucleotide variants (SNVs) in functionally important regions of the proteins within certain groups of chickens were detected, in particular the European breeds and indigenous birds from Ethiopia and Nigeria. In addition, we also found that two out of four SNVs located in the chIFITM1 (Ser36 and Arg77) and chIFITM3 (Val103) proteins were simultaneously under positive selection. Together these data suggest that IFITM genetic variation may contribute to the capacities of different chicken populations to resist viral infection. 
Type Of Material Model of mechanisms or symptoms - non-mammalian in vivo 
Year Produced 2019 
Provided To Others? Yes  
Impact This paper is currently in press in. BMC Genomics. 
 
Description Horizon Discovery Ltd Cambridge Research Park, United Kingdom 
Organisation Horizon Discovery Group plc
PI Contribution Verify IFITM knock-out cell by qPCR and immuno-blotting. Verification of further knock-out cell lines by DNA sequencing, immuno-blotting and qPCR Cell infection with Avian Influenza Viruses Assess the viral titre of cells infected with Influenza Viruses in K/O and wildtype cells. Quantitative analysis of viral infection in IFITM K/O cells Demonstrate increased AIV viral titres at lab scale and estimation of what this may equate to with respect to vaccine dose yields. Analysis of IFITM K/O cell lines permissivity to a range of additional animal viruses (Relevant to the LVIF). Milestone 7: Demonstrate permissivity increased and viral titres for a range of LVIF relevant viruses in the edited cell lines.
Collaborator Contribution Develop a CRISPR/cas9 gene editing system to specifically target the chIFITM locus in avian cell culture
Impact Grant funding
Start Year 2017
 
Description ILRI Research Collaboration (Kenya, March 2017) 
Organisation International Livestock Research Institute (ILRI)
Country Kenya 
Sector Charity/Non Profit 
PI Contribution Exchange of ideas and samples for indigenous breeds of livestock
Collaborator Contribution Contributed samples for analysis.
Impact Analysis of indigenous breed genomic sequences for Immune function genes.
Start Year 2017
 
Description Industrial partnership-Aviagen 2018 
Organisation Aviagen Group
Country United States 
Sector Private 
PI Contribution We have established a significant industrial partnership with Aviagen group Ltd. The nature of this work remains confidential.
Collaborator Contribution The nature of this work remains confidential.
Impact The nature of this work remains confidential.
Start Year 2018
 
Description Research Collaboration with Nigeria (23rd - 28th January 2017) 
Organisation University of Nigeria
Country Nigeria 
Sector Academic/University 
PI Contribution Analysis of rare breed indigenous chicken in Nigeria.
Collaborator Contribution Helped with sampling and background information of the breed.
Impact Analysis is ongoing.
Start Year 2017
 
Title Chicken IFITMs 
Description The present invention provides an avian cell in which the expression or activity of one or more of the following genes, or a homologue thereof: Chicken IFITM1 (SEQ ID No. 1); Chicken IFITM2 (SEQ ID No. 2) and Chicken IFITM3 (SEQ ID No. 3) is reduced. The invention also provides methods for passaging viruses in avian cells, embryos and/or avian cell lines which have reduced expression of one or more IFITM genes and methods which involve investigating the sequence of one or more of the following genes, or a homologue thereof: Chicken IFITM1 (SEQ ID No. 1); Chicken IFITM2 (SEQ ID No. 2) and Chicken IFITM3 (SEQ ID No. 3). 
IP Reference PCT/GB2014/051693 
Protection Patent granted
Year Protection Granted 2018
Licensed Commercial In Confidence
Impact The present invention provides an avian cell in which the expression or activity of one or more of the following genes, or a homologue thereof: Chicken IFITM1 (SEQ ID No. 1); Chicken IFITM2 (SEQ ID No. 2) and Chicken IFITM3 (SEQ ID No. 3) is reduced. The invention also provides methods for passaging viruses in avian cells, embryos and/or avian cell lines which have reduced expression of one or more IFITM genes and methods which involve investigating the sequence of one or more of the following genes, or a homologue thereof: Chicken IFITM1 (SEQ ID No. 1); Chicken IFITM2 (SEQ ID No. 2) and Chicken IFITM3 (SEQ ID No. 3). This patent has enabled us to leverage funding for continued work.
 
Description Chair of organising Committee for Wellcome Trust-Animal Genetics and Diseases 2017 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact This meeting brought together specialists working on the interface between genomics, genetic engineering and infectious disease with the aims of improving animal and human health and welfare.

Scientific sessions included:
Genetics of immune responses and disease resistance
Genetically engineered livestock (including genome editing)
Quantitative genetics and epigenetics applied to disease
Epidemiology and pathogen evolution
Bioinformatics, comparative and functional genomics
Precision medicine of animal companions
Year(s) Of Engagement Activity 2017
URL https://coursesandconferences.wellcomegenomecampus.org/events/item.aspx?e=635&dm_i=2SUU,HOGH,4R4AW1,...
 
Description Encouraging women into science and engineering STEM. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact MH Visited Warwick School, Redhill to talk to secondary students about careers in STEM. A very positive outcome and well received.
Year(s) Of Engagement Activity 2018
 
Description Holt School Surrey. Bee meadow & A-level outreach Talk. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact I discussed the importance of pollinators for biodiversity and food security with the younger students. I also discussed careers in science with A-Level students. There was good interaction and many questions surrounding both topics.
Year(s) Of Engagement Activity 2018
 
Description Kingdown School Warminster Wiltshire talk to 6th form students about career in science. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact I presented my career path to the students to engage them on STEM careers. I had a good level of discussion and many questions.
Year(s) Of Engagement Activity 2018
 
Description Organiser of the Animal Genetics and Diseases conference 08 - 10 May 2019 Wellcome Genome Campus, UK. Highlighting recent advances in animal genetics and genomic technologies. 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact I am on the organising committee for this conference. The second conference in this series will highlight recent advances in animal genetics and genomic technologies. It will bring together specialists working on the interface of genomics, genetic engineering and infectious disease with the aims of improving animal and human health and welfare.

Novel genomic technologies, mathematical modelling and quantitative genetics approaches, applied to host animals, as well as their pathogens, have transformed the understanding of animal diseases, host-pathogen interactions and epidemiology and their effects on productivity of farmed animal species and food supply chains.

This year's conference will not only put the spotlight on the immune response of host animals and epidemiology but also cover the genetics and genomics of pathogens and the impact of animal-human relationships.

We encourage registrations from researchers, breeders and technical specialists interested in learning and disseminating the latest cutting-edge techniques and methodologies across model species, wildlife, farmed animals and companion animals.
Year(s) Of Engagement Activity 2019
URL https://coursesandconferences.wellcomegenomecampus.org/our-events/animal-genetics-diseases-2019/?dm_...
 
Description Taiwan -UK Partnering Award: Surveillance of influenza viruses 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Attended the International Flu Virus Symposium for Celebrating IPM 47th Anniversary" in Tri-Service General Hospital in Taipei, Taiwan.

Tri-Service General Hospital address:
No. 325, Sec. 2, Chenggong Rd., Neihu Dist., Taipei City 114, Taiwan (R.O.C.)

Invited speaker for medical staff and research scientists.
Year(s) Of Engagement Activity 2019
URL https://www.cdc.gov.tw/rwd/english
 
Description Winston Churchill Careers Fair. Winston Churchill School, Hermitage Road, St Johns, Woking, Surrey, GU21 8TL 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact AS attended and presented at this event to stimulate increased interest in science and research.
Year(s) Of Engagement Activity 2018