UK-China partnership on Global Food Security: Combating avian tumor diseases for sustainable poultry production
Lead Research Organisation:
The Pirbright Institute
Department Name: Avian Oncogenic Viruses
Abstract
Infectious diseases result in direct and indirect losses at various steps of poultry farming and amongst them neoplastic disease caused by viruses is a major economic problem faced by the poultry industry worldwide. The oncogenic viruses causing neoplastic infection in chickens are herpesviruses comprising of Marek's disease virus (MDV) and retroviruses comprising of reticuloendotheliosis virus (REV) and avian leucosis virus (ALV). ALV is the most common naturally occurring avian retrovirus that can cause a variety of neoplastic disease conditions in chickens. In addition to causing neoplasia, ALV is known to be associated with reduced productivity and other production problems in affected flocks. Based on the properties of viral envelope glycoproteins, ALV is classified into six subgroups: A, B, C, D, E and J. Originally isolated by the Pirbright Institute in the late 1980s, ALV-J spreads widely and induces myeloid leukosis and a broad spectrum of additional disorders, such as histiocytic sarcoma, hemangiomas and erythroblastosis. The original strains have almost been eradicated from most of the commercial poultry flocks in Europe and the United States but in Asia, the virus evolved into a great variety and recently emerging Chinese isolates coarsely induce various tumours in both commercial laying hen flocks as well as native Chinese breeds of chickens. Defining the molecular background of the tumorigenesisis is needed to develop more effective control and eradication programmes in China. In this proposal, we aim to obtain detailed comprehensive picture on the molecular events to understand and predict the molecular pathways to ALV-J induced cancer. This includes non-coding RNA profiling for the role of non-coding RNAs in ALV-J induced oncogenesis, oncogene myc targetome identification and the role of myc-binding in maintaining the transformed phenotype and myc knockout by CRISPR/Cas9 system for the role of myc in maintaining the transformed phenotype. As the co-infection of ALV-J with other tumor viruses such as Marek's disease virus and Reticuloendotheliosis virus is very common and caused more serious pathogenic effects, we are going to elucidate the synergistic mechanism of ALV-J+REV/ALV-J+MDV co-infection in vitro and in vivo. The findings will be very valuable in developing new approaches for the eradication of ALV-J in China and control of cancers caused by oncogenic viruses. We also want to establish the resistant cell line by editing ALV-J receptor NHE1 using CRISPR/Cas9 system to explore genetic resistant to ALV-J. Finally, we will develop the rapid diagnostic kits to detect ALV-J infection in the field to aid the ALV-J control and eradication in China.
Planned Impact
Poultry industry is a rapidly growing sector crucial for the global food security, acting as a major source of protein for the growing world population. Avian leukosis viruses (ALV) are retroviruses associated with the induction of tumours of haemopoietic cells in birds. Although diseases due to ALV have largely been controlled in Europe and USA through successful eradication programmes, it continues to be a major problem in many countries including China, where it has caused serious economic losses in recent years. In particular, myeloid leukosis and other tumours due to ALV-J remains a major disease that threatens the Chinese poultry industry. Detailed understanding of the molecular basis of tumours induced by ALV-J, generation of resistant cell line, and development of rapid diagnostic tools for ALV-J detection in the field all will benefit development of new strategies for control and eradication of ALV-J in Chinese poultry industry. The beneficiaries of this research will include academic scientists, the poultry breeding companies, chicken farms, diagnostic professionals, the Pirbright Institute, the BBSRC and its stakeholders such as Defra and the UK poultry farming industry.
The research will have general impact with the wider scientific community, veterinary and medical practitioners, students and general public. Engagement with these diverse groups will be achieved via meetings, articles in the trade press, tailored web pages, press releases to the media and outreach events in schools.
In the longer term the research may lead to medical benefits by improving control of human virus pathogens which will benefit the UK MRC and UK department of Health, the pharmaceutical industry and international organization such as the World Health Organization.
If the proposed studies lead to new approaches for controlling ALV-J or other viral diseases, additional funding will be sought from relevant funding agencies and other sources for further development. There is extensive experience within the Pirbright Institute of patent applications and commercialisations, new opportunities will feed into an established system for technology development and knowledge transfer by the Pirbright Business Development group.
Rapid-onset transformation induced by myc is also a good model for studying cancers in other species including humans as myc is one of the most frequently deregulated oncogenes in human malignancies.
The research will have general impact with the wider scientific community, veterinary and medical practitioners, students and general public. Engagement with these diverse groups will be achieved via meetings, articles in the trade press, tailored web pages, press releases to the media and outreach events in schools.
In the longer term the research may lead to medical benefits by improving control of human virus pathogens which will benefit the UK MRC and UK department of Health, the pharmaceutical industry and international organization such as the World Health Organization.
If the proposed studies lead to new approaches for controlling ALV-J or other viral diseases, additional funding will be sought from relevant funding agencies and other sources for further development. There is extensive experience within the Pirbright Institute of patent applications and commercialisations, new opportunities will feed into an established system for technology development and knowledge transfer by the Pirbright Business Development group.
Rapid-onset transformation induced by myc is also a good model for studying cancers in other species including humans as myc is one of the most frequently deregulated oncogenes in human malignancies.
Publications
Cui X
(2022)
TMT-based proteomic analysis reveals integrins involved in the synergistic infection of reticuloendotheliosis virus and avian leukosis virus subgroup J.
in BMC veterinary research
Zhao W
(2020)
The Isolation and Molecular Characterization of an Astrovirus From "Yellow" Chickens, China
in Frontiers in Veterinary Science
Zhang J
(2021)
Synergistic pathogenesis of chicken infectious anemia virus and J subgroup of avian leukosis virus.
in Poultry science
Zhou D
(2018)
Reticuloendotheliosis virus and avian leukosis virus subgroup J synergistically increase the accumulation of exosomal miRNAs
in Retrovirology
Qiao D
(2021)
Regulation of Avian Leukosis Virus Subgroup J Replication by Wnt/ß-Catenin Signaling Pathway.
in Viruses
Roy S
(2022)
Proviral ALV-LTR Sequence Is Essential for Continued Proliferation of the ALV-Transformed B Cell Line.
in International journal of molecular sciences
Xu M
(2021)
Novel mutation of avian leukosis virus subgroup J from Tibetan chickens.
in Poultry science
Description | In situ CRISPR/Cas9 gene editing tools for functional genomics in ALV-transformed cell lines: ALV is a typical retrovirus which induces the pathogenicity by integration into the host genome. After integration, LTR acts as promoter to activate the expression of the oncogene and induce tumors. The development of lymphoid leukosis is a result of cooperation of c-Myc and c-Bic (encoding the oncogenic miRNA mir-155) integration and activation by ALV. Using the CRISPR/Cas9 editing system, we have shown that both LTR and c-Myc but not miR-155 are essential for the continued proliferation of ALV transformed B cell line HP45. The role of c-Myc oncogene in regulating the transcriptional machinery of ALV transformed cells: c-Myc is a transcription factor that binds to specific signature sequences in the different promoters in regulating expression. To examine the role of Myc in ALV-induced transformation, we determined the global Myc-binding sites in the genome of ALV cell line HP45 using Myc-ChIP (chromatin immunoprecipitation) analysis after tagging c-Myc with AM tag which has got a good Chippable antibody. After the ChIP-seq assay, a complete data set of the myc binding sites in the whole genome of HP45 was obtained. Speci?c high-sensitivity enzymatic reporter unlocking (SHERLOCK) platform for diagnosis of ALV: As vaccines are not available for ALV, the current practice of eradication involves early detection and removal of infected birds to reduce contact and the incidence of horizontal spread. To aid the eradication program, rapid, sensitive, specific, easy-to-use and cost effective on-site diagnostic method is needed. We have developed a lateral flow detection method using Cas13a-based SHERLOCK platform for rapid detection of the most common ALV subgroups A, B, and J. |
Exploitation Route | Establishment of the pipeline for in situ CRISPR editing of lymphoma-derived cell lines, gives the opportunity for the identification of critical viral and host genes involved in features such as virus-host interactions, neoplastic transformation, and virus latency. Determination of the global myc-binding profiles in transformed macrophages could provide insights into its role in transformation. Achieving rapid detection of ALV infection is imperative in effective control of the spread of ALVs. |
Sectors | Agriculture, Food and Drink |
Description | BBSRC IAA The Pirbright Institute |
Amount | £300,000 (GBP) |
Funding ID | BB/S506680/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 03/2018 |
End | 03/2021 |
Description | CRISPR/Cas system-based molecular diagnostics for avian viral pathogens |
Amount | £21,263 (GBP) |
Organisation | The Pirbright Institute |
Sector | Academic/University |
Country | United Kingdom |
Start | 01/2020 |
End | 03/2020 |
Description | Collaborative partnership for establish of PhysioMimix™ OOC system, part of Pirbright Institute Flexible Talent Mobility Account |
Amount | £180,000 (GBP) |
Funding ID | BB/S507945/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 09/2020 |
End | 12/2020 |
Description | Development of HVT vectored vaccine |
Amount | £100,000 (GBP) |
Organisation | MSD Animal Health |
Sector | Private |
Country | United Kingdom |
Start | 11/2022 |
End | 04/2023 |
Description | Development of an HVT triple insert vaccine using CRISPR/Cas9 technology and novel vector vaccines |
Amount | £140,000 (GBP) |
Organisation | MSD Animal Health |
Sector | Private |
Country | United Kingdom |
Start | 02/2019 |
End | 01/2020 |
Description | International Exchanges 2021 Cost Share (NSFC) |
Amount | £12,000 (GBP) |
Funding ID | IEC\NSFC\211090 |
Organisation | The Royal Society |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 04/2022 |
End | 03/2024 |
Title | Lateral flow detection of avian leukosis viruses and Reticuloendotheliosis virus |
Description | The system for detecting ALV/REV based on CRISPR-Cas13a, a SHERLOCK reaction system consists of specific RPA primers for amplifying target nucleic acid fragments, Cas13a protein, crRNA, and Lateral flow chromatography test strip for visualized display of results. The assay started with pre-amplification of either a DNA or RNA target input. Amplified targets are then converted to RNA via T7 transcription and detected by Cas13-crRNA complexes, which activate and cleave fluorescent RNA reporters based on its collateral cleavage nature. The result can be visualized using colorimetric lateral flow reaction. The test is rapid, sensitive and specific for detection of ALV/REV at 37°C. The product can commercialized with a great clinical application prospects. |
Type Of Material | Technology assay or reagent |
Year Produced | 2020 |
Provided To Others? | No |
Impact | The early identi?cation and removal of virus-shedding birds are essential to reduce the transmission. This rapid, simple-to-use and cost effective on-site diagnostic method will aid the eradication of ALV/REV in the affected area. |
Description | Professor Aijian Qin & Kun Qian |
Organisation | Yangzhou University |
Department | Faculty of Veterinary Medicine |
Country | China |
Sector | Academic/University |
PI Contribution | Provided knowledge, reagent and information on ALV research |
Collaborator Contribution | Provided samples and reagent for MDV and ALV research |
Impact | 5 publications have been generated: 1. Zhao W, Wu Z, Yao Y, Qian K. The Isolation and Molecular Characterization of an Astrovirus from "Yellow" Chickens, China. Frontiers in Veterinary Science, 27 October 2020 | https://doi.org/10.3389/fvets.2020.581862. 2. Yang F, Feng C, Yao Y, Qin A, Shao H and Qian K. Antiviral effect of baicalin on Marek's disease virus in CEF cells. BMC Veterinary Research. 2020, 16:371 3. Zhou X, Wang L, Shen A, Shen X, Xu M, Qian K, Shao H, Yao Y, Nair V, Ye J, Qin A. Detection of ALV p27 in cloacal swabs and virus isolation medium by sELISA. BMC Vet Res. 2019 Oct 30;15 (1):383. doi: 10.1186/s12917-019-2150-z. 4. Qian K, Tian X, Shao H, Ye J, Yao Y, Nair V, Qin A. Identification of novel B-cell epitope in gp85 of subgroup J avian leukosis virus and its application in diagnosis of disease. BMC Veterinary Research. 2018 Sep 26;14(1):295. doi: 10.1186/s12917-018-1622-x. 5. Qian K, Cheng X, Zhang D, Shao H, Yao Y, Nair V, Qin A. Antiviral effect of lithium chloride on replication of avian leukosis virus subgroup J in cell culture. Arch Virol. 2018 Jan 11. doi: 10.1007/s00705-017-3692-7. |
Start Year | 2018 |
Description | Professor Ziqiang Cheng |
Organisation | Shandong Agricultural University |
Country | China |
Sector | Academic/University |
PI Contribution | Provided knowledge and information on ALV research |
Collaborator Contribution | Provided samples and reagent for ALV research |
Impact | $4 publication have been generated: 1. Pang, Y., Zhou, D., Xue, J., Zhou, J., Zhang, Y., Zheng, G., Yuan, S., Yao, Y. and Cheng, Z. Interplay between CTHRC1 and the SU protein of avian leukosis virus subgroup J (ALV-J) facilitates viral replication. Virus Research. 2019. https://doi.org/10.1016/j.virusres.2019.02.014 2. Zhu, M., Zhou, J., Ma, X., Li, G., He, S., Tang, H., Yao, Y. and Cheng, Z. CCCH-type zinc finger antiviral protein is specifically overexpressed in spleen in response to subgroup J avian leukosis virus infection in chicken. Research in Veterinary Science. 2019. 123: 65-70. 3. Zhou J, Zhao GL, Wang XM, Du XS, Su S, Li CG, Nair V, Yao YX, Cheng ZQ. Synergistic Viral Replication of Marek's Disease Virus and Avian Leukosis Virus Subgroup J is Responsible for the Enhanced Pathogenicity in the Superinfection of Chickens. Viruses. 2018 May 18;10(5). pii: E271. doi: 10.3390/v10050271. 4. Zhou D, Xue J, He S, Du X, Zhou J, Li C, Huang L, Nair V, Yao Y, Cheng Z. Reticuloendotheliosis virus and avian leukosis virus subgroup J synergistically increase the accumulation of exosomal miRNAs. Retrovirology. 2018 Jul 3;15(1):45. doi: 10.1186/s12977-018-0427-0. |
Start Year | 2018 |
Description | Yulong Gao |
Organisation | Chinese Academy of Agricultural Sciences |
Department | Harbin Veterinary Research Institute (HVRI) |
Country | China |
Sector | Public |
PI Contribution | Provided knowledge, reagent and information on ALV research |
Collaborator Contribution | Provided samples and reagent for MDV and ALV research |
Impact | Two publications have been generated: 1. Li K, Liu Y, Xu Z, Zhang Y, Yao Y, Nair V, Liu C, Zhang Y, Gao Y, Qi X, Cui H, Gao L, Wang X. Prevention of Avian Retrovirus Infection in Chickens Using CRISPR-Cas9 Delivered by Marek's Disease Virus. Molecular Therapy - Nucleic Acids 2020 Sep 4; 21:343-353. doi: 10.1016/j.omtn.2020.06.009. 2. Ren C, Xie R, Yao Y, Yu M, Chang F, Xing L, Zhang Y, Liu Y, Wang S, Farooque M, Wang Y, Qi X, Liu C, Zhang Y, Cui H, Li K, Gao L, Pan Q, Nair V, Wang X, Gao Y. MiR-125b Suppression Inhibits Apoptosis and Negatively Regulates Sema4D in Avian Leukosis Virus-Transformed Cells. Viruses. 2019 Aug 7;11(8). pii: E728. doi: 10.3390/v11080728. |
Start Year | 2018 |
Description | AN INTERVIEW WITH PROFESSOR VENUGOPAL NAIR |
Form Of Engagement Activity | Engagement focused website, blog or social media channel |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Professor Venugopal Nair is a Research Group Leader at The Pirbright Institute, a visiting Professor of Avian Virology at the Department of Zoology, and a Jenner Investigator at the Jenner Institute, University of Oxford. He is also a member of the Microbiology Society, and in this interview, he tells us more about his research into viral diseases of livestock. |
Year(s) Of Engagement Activity | 2020 |
URL | https://microbiologysociety.org/membership/meet-our-members/focus-area-viruses/an-interview-with-pro... |
Description | Interview by CGTN |
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 | Increased profile of both Institute and The UK-China Centre of Excellence for Research on Avian Diseases (CERAD) |
Year(s) Of Engagement Activity | 2019 |
URL | https://newseu.cgtn.com/news/2020-01-28/British-and-Chinese-scientists-join-forces-to-fight-avian-di... |
Description | Invited talk on 28th September 2020 on ''Challenges and opportunities for control of infectious animal diseases including zoonosis' |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | International Webinar of 'Epistemological approaches to animal disease control programmes with special reference to rabies' |
Year(s) Of Engagement Activity | 2020 |
Description | School visit (Tilingborne) |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | Stimulating increased interest in science and research |
Year(s) Of Engagement Activity | 2019 |
Description | Science Festival (Cheltenham) |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Stimulating increased interest in science and research |
Year(s) Of Engagement Activity | 2019 |