Primary Supervisor: Dr Anne Straube, Warwick Medical School
Secondary supervisor: TBC
PhD project title: Cell-to-cell spread of Marek’s Disease Virus
University of Registration: University of Warwick
The sustainability of global food production is under threat from the increase in human population, reduced availability of natural resources and climate change. With a current global production of 55 billion chickens annually, poultry meat and eggs dominate animal protein products for human consumption world-wide. The UK poultry industry, contributing around £3.5 billion to the economy annually, provides more than 90% of the poultry meat and egg for the country. Avian diseases, particularly those induced by viral pathogens, remain a major threat for sustainable poultry production. Marek's disease is one of the most widespread and economically important diseases of poultry, with global losses estimated to more than US $2,000 million. MD is a fatal, highly contagious, neoplastic disease affecting chickens. It is caused by an alphaherpesvirus called Gallid herpesvirus-2 or Marek's Disease Virus (MDV). Advancing our understanding of MDV by identifying and characterising its life cycle and virus-host interactions will enable us to develop interventions that limit the transmission of MDV. MDV is a large DNA virus that is strictly cell-associated. Thus live infected cells have to be used to maintain the infectivity of the virus and it is highly dependent on the host cytoskeleton to transport MDV from entry site to nucleus, and new virus particles via Golgi back to the cell surface to transmit to neighbouring cells. How the virus is transferred from one cell to the other is not clear. It is assumed that this occurs through the formation of tunnelling nanotubes (TNTs) that connect the cytoplasm of neighbouring cells and allow passage of viruses. The aim of the project will be to directly observe transfer of the virus between cells and to identify host cell factors and viral proteins involved in orchestrate this process, i.e. trigger the formation of TNTs, and transporting the virus towards and through TNTs.
Thus the project will combine live cell imaging using lattice light sheet microscopy, quantitative image analysis and particle tracking,
References: (there is no published information on the mechanism of MDV cell-to-cell transport yet, but from other viruses and prions)
- Gousset K, Schiff E, Langevin C, Marijanovic Z, Caputo A, Browman DT, Chenouard N, de Chaumont F, Martino A, Enninga J, Olivo-Marin JC, Männel D, Zurzolo C. Prions hijack tunnelling nanotubes for intercellular spread. Nat Cell Biol. 2009 Mar;11(3):328-36. doi: 10.1038/ncb1841. Epub 2009 Feb 8. PMID: 19198598.
- Sherer NM, Lehmann MJ, Jimenez-Soto LF, Horensavitz C, Pypaert M, Mothes W. Retroviruses can establish filopodial bridges for efficient cell-to-cell transmission. Nat Cell Biol. 2007 Mar;9(3):310-5. doi: 10.1038/ncb1544. Epub 2007 Feb 11. PMID: 17293854; PMCID: PMC2628976.
- Roberts KL, Manicassamy B, Lamb RA. Influenza A virus uses intercellular connections to spread to neighboring cells. J Virol. 2015 Feb;89(3):1537-49. doi: 10.1128/JVI.03306-14. Epub 2014 Nov 26. PMID: 25428869; PMCID: PMC4300760.
BBSRC Strategic Research Priority: Sustainable Agriculture and Food: Animal Health and Welfare & Understanding the Rules of Life:Microbiology
Techniques that will be undertaken during the project:
Animal cell culture; molecular biology to clone cDNAs and introduce deletions and point mutations into expression constructs; RNAi and DNA transfection to deplete proteins of choice and supplement with wild-type and mutant versions; proteomics to identify interacting proteins; protein purification and protein-protein interaction analysis; live cell imaging assays (e.g. imaging infected chicken cells using lattice light sheet and spinning disk confocal microscopy); total internal reflection fluorescence microscopy paired with biochemical reconstitution assays; image and statistical data analysis using ImageJ and Matlab.
Contact: Dr Anne Straube, University of Warwick