Project 3

Lead supervisor: Prof Len Seymour, Department of Oncology, University of Oxford

Co-supervisor: Prof Simon Draper, Jenner Institute, University of Oxford

Commercial partner: Oxford Genetics Ltd, Oxford Science Park

This project will harness the power of genetically modified memory B cells (GMBcs) to proliferate and secrete chosen monoclonal antibodies. This will provide a new healthcare platform, covering both prophylaxis and therapy, with applications across infection and immunity (e.g. malaria, HIV, influenza), cancer (e.g. antibodies targeting cancer antigens) and rheumatoid arthritis (e.g. induction of anti-TNF). Patients will receive a semi-synthetic autologous immune component producing monoclonal antibodies with proven biological activity.

There will be three main components:

(i)                  Proof of principle.

1. Gene delivery: B cells resist standard lentiviral transduction but can be infected with GALV-pseudotyped vectors. This system will be validated using a B cell line and primary B cells and range of alternative glycoproteins will be screened for optimal infectivity.
2. B cell genome engineering: Next generation CRISPR design algorithms will be used to develop efficient knock-out strategies to remove endogenous BCR (B cell receptor) expression from a B cell line and replace it with a simple model monoclonal antibody recognising Her2 (trastuzumab).
3. Response: GMBcs will be triggered to stimulate B cell division and trastuzumab secretion. B cell biology and antigen-triggered responses will be compared to WT responses through proteomics and transcriptomics, and antibody potency will be measured in a proliferation-inhibition assay.

(ii)                Prototype development - human primary cells. Memory B cells will be isolated from PBMCs and engineered to replace endogenous BCR with trastuzumab. We will also develop a prototype antimalarial agent, by encoding R5.016 – the most potent known monoclonal antibody against blood-stage P. falciparum.

(iii)               Malaria prophylaxis in vivo. We will encode an anti-malaria monoclonal antibody (2A10) into murine primary B cells and assess their ability to protect mice against malaria-invading sporozoites in a challenge model using a P. berghei rodent malaria parasite transgenic for the P. falciparum CSP antigen.