Dunn School researchers join the fight against the novel Coronavirus
Dunn School researchers join the fight against the novel Coronavirus
Research groups in the Dunn School join the international battle against the novel Coronavirus, SARS-CoV-2. While some groups work on developing cutting edge diagnostic tools, others work relentlessly on identifying new targets for drug development.
Below are detailed reports on the ongoing efforts.
The Sattentau lab tackles Coronavirus research from three separate angles
The Sattentau lab has made recombinant soluble SARS-CoV2 spike protein, the major surface protein used by the virus to infect host cells, and have demonstrated it can be detected by immune sera, anti-Spike monoclonal antibodies and soluble ACE2 ligands. This protein can now be used to detect anti-Spike antibodies produced by infected people, and in animal models following immunisation. The lab intend to use their findings in an already established pipeline to use crosslinked Spike protein as a potential vaccine candidate.
In order to study SARS-CoV2 cell infection, Maeva Dupont (Postdoctoral researcher, Sattentau lab) has been working on setting up immunofluorescence labelling of infected cells with help and support from Prof. Ilan Davies (Department of Biochemistry, University of Oxford) and James Felce (Oxford Nanoimaging).
Additionally, Rebecca Moore (Lab manager, Sattentau lab) has been helping Prof. William James set up a containment level 3 (CL3) lab so that work on SARS-CoV-2 can take place in the Dunn School. Prior to the pandemic the CL3 suite was used solely for HIV-1 work, and none of the procedures were compatible with an air-borne pathogen. Having received HSE approval to start work, William and Rebecca began by making batches of the strains of SARS-CoV-2 from Australia and England, and training other researchers from within the Dunn School and across the University of Oxford to work on the virus.
Currently the CL3 facility in the Dunn School is being used to:
1) Test panels of antivirals and monoclonal antibodies sent from laboratories around the world to determine if they have any anti-SARS-CoV-2 activity
2) Check plasma samples from patients convalescent from SARS-CoV-2 to prove that, despite containing low level viral RNA, these samples aren’t infectious
3) Undertake a number of research projects that have been approved by the oversight committee set up by Prof. William James at the beginning of the pandemic to manage the large number of requests to use the facility
The Sattentau lab would like to thank the following:
From the Dunn School: Prof. William James, Wayne Swan, Peter Stroud, Marcus Bottomley and Bruce Coppen from workshop, John Marriott from IT, Alan Wainman from Jordan Raff’s lab and Prof. Herman Waldmann who made the soluble ACE2 ligands. Prof. Alain Townsend (MRC Weatherall Institute of Molecular Medicine), Prof. Gavin Screaton (University of Oxford Medical Sciences Division) and Prof. Jim Naismith (Rosalind Franklin Institute) for anti-Spike antibodies. Linda Randall from the Pathology Support Building (PSB) for help in accommodating all the new people coming into the building, and other members of the PSB staff for dealing with the increased work load in the facility. Tracey Mustoe from the safety office at the University of Oxford for help in approving the CL3 lab. The British Clinical Services for being instrumental in the initial recommissioning of the CL3 suite. Prof. Robin Shattock from Imperial College for providing the SARS-CoV2 plasmid.
Written by Isabella Maudlin
Dunn School Researchers Establish Core Facility for Study of Coronavirus
In respons to the coronavirus pandemic, a SARS-CoV-2 facility has been established by a group of Dunn School researchers, with collaborative projects getting underway from May 2020.
William James and colleagues established this core facility to enable the culture and study of the SARS-CoV-2 virus. Through this endeavour, they are now involved in a series of collaborative projects involving scientists from across the University. These projects are being driven forward by a dedicated team including Becky Moore, Javier Gilbert, Michael Knight, Tom Beneke, Louis Xu Liu, Adam Harding, Alun Vaughan-Jackson, Oliver Sampson, Cathy Browne and Sally Cowley.
Since the new containment facility enables researchers to culture the virus in cells, current projects include the testing of potential antivirals: drugs that may act to inhibit the virus once it has entered the host cell. Antibodies are also being tested to see if these can neutralise the coronavirus, binding to the virus to prevent it from entering cells in the first place. Alongside this, researchers have been analysing a range clinical samples, such as patient bloods, to test whether the small proportion of them that contain coronavirus RNA are infectious. The group found that these samples were unable to infect cells in culture, suggesting that they were safe to handle. This finding should enable the samples to be handled with confidence, thus increasing the speed and ease of diagnosis in medical settings.
Professor James’ group usually focuses on how viruses such as HIV are able to enter and replicate in macrophages. His lab makes use of human induced pluripotent stem cells (iPSCs), which can be reprogrammed to generate a range of different cell types. In the James lab, iPSCs are differentiated to produce models of macrophages and their neighbouring cells. The new facility has allowed them to apply this expertise in tackling SARS-CoV-2, with Sally Cowley developing protocols for the differentiation of lung epithelial-like cells. This will provide a basis for investigating crosstalk between different cell types during coronavirus infection.
By enabling the culture of SARS-CoV-2, the new facility has opened up a huge range of possibilities for coronavirus-related research that is already having a direct impact during this pandemic.
Written by Laura Hankins
Dunn School Researchers Set Their Sights on the Coronavirus RNA Polymerase
As part of global efforts against the coronavirus pandemic, researchers at the Dunn School are searching for antivirals that target the SARS-CoV-2 polymerase. Such antivirals could prove crucial to developing a treatment for the disease, which currently has no known cure.
Ervin Fodor’s lab normally works on the influenza virus, focusing on its RNA polymerase. Polymerases are enzymes that are required for the replication of the viral genome and therefore play a key role in the production of new viruses during infection. In response to the threat posed by COVID-19, the Fodor Lab have turned their attention to the RNA polymerase of novel coronavirus SARS-CoV-2 and are studying how drugs could be applied to inhibit its function.
To achieve this, the lab is purifying the coronavirus polymerase as well as its cofactors. Drug screening can then be carried out in order to identify antivirals that might be effective inhibitors of this RNA polymerase. Structural approaches are also being employed to compare the structure of the polymerase with and without the presence of these inhibitory drugs. These structural biology studies are in collaboration with Professor Jonathan Grimes at the Division of Structural Biology (STRUBI), University of Oxford, a long-term collaborator of the Fodor group. They are also working closely with Dr Aartjan te Velthuis at the University of Cambridge.
Early results have already been released as a preprint, with the researchers reporting that the small molecule enisamium acts as an inhibitor of both the influenza and the SARS-CoV-2 RNA polymerases. In addition to the clear therapeutic potential of this discovery, the findings may also have broader implications for the application of this treatment against other viral polymerases.
Written by Laura Hankins
To read more about the early findings of the Fodor lab’s work, see this preprint on bioRxiv: https://www.biorxiv.org/content/10.1101/2020.04.21.053017v2
Targeting macrodomain enzyme in coronaviruses to inhibit viral infection
University of Oxford is currently an epicentre for cutting-edge research and development related to the current coronavirus (COVID-19) pandemic. A research group at Dunn School, led by Ivan Ahel, has joined the global fight against SARS-CoV-2, the virus causing COVID-19. They plan to find new inhibitors against the enzyme macrodomain ADP-ribosyl hydrolase, a new target in SARS-CoV-2, essential for viral replication in human cells.
ADP-ribose is covalently attached to macromolecules such as proteins, by the enzyme poly-ADP-ribose polymerases (PARPs) which has been linked to various important cellular processes such as DNA repair and host-virus interactions. Many PARPs are known to support the host defence response to viruses but ingeniously, coronaviruses encode macrodomain enzymes, which remove ADP-ribose from antiviral proteins to counter the host innate immune reaction to virus infection.
The group plans to undertake several independent in vitro and in silico screens to uncover potential inhibitors against both highly similar SARS-CoV-1 (coronavirus that caused the 2003 SARS epidemic) and SARS-CoV-2 macrodomains. Preliminary work has started to generate crystals for these proteins needed for the XChem screens, which will be done at Diamond Light Source. Ivan Ahel hopes this effort will lead to the development of potential new antiviral drugs, targeting viral macrodomains of coronaviruses such as COVID-19.
Written by Iqbal Dulloo