MRC DiMeN Doctoral Training Partnership: Fine-tuning blood cell development: Using antibodies as surrogate cytokines to modify receptor activation and signalling in haematopoiesis

Background

Cytokines and growth factors are essential for many aspects of mammalian homeostasis, and are fundamental regulators of haematopoiesis, controlling blood cell development, haematopoietic stem cell (HSC) maintenance and immunity. Given the significant structural and functional diversity of the cytokine superfamilies, it is remarkable that just two cytokines, erythropoietin (EPO) and thrombopoietin (TPO) are almost entirely responsible for the production of >99% of circulating blood cells, with millions of erythrocytes and platelets generated every second. Consequently, aberrant EPO and TPO signalling can have profound pathological effects on haematopoiesis, ranging from aplastic anaemia to haematological malignancies. Using a combination of single molecule super resolution microscopy and structure/function analyses, we have recently uncovered a new mechanistic paradigm in cytokine signalling whereby TPO and EPO receptors exist at the cell surface as monomers, rather than as pre-formed dimers, and dimerize in response to their cognate ligands (Wilmes et al., Science, 2020). Not only did these findings highlight the significant complexity of receptor activation, they provide us with the opportunity to manipulate these receptors, precisely control signalling outcomes and fine-tune blood cell development using antibody-based surrogate cytokines (Cui et al., PNAS, 2021). These agents can now be further developed to manipulate haematopoiesis in various disease states by altering lineage differentiation, differentiation and HSC maintenance.  

Objectives

1.        Identify antibodies and antibody fragments that interact with the TPO and EPO receptors.

2.        Characterise their functionality as surrogate cytokines; monitoring receptor activation, downstream signalling and haematopoiesis. 

Project Outline

This iCASE project will focus on the identification, development and functional characterization of monoclonal antibodies and antibody fragments as surrogate cytokines for EPO and TPO. You will be primarily based in the Department of Biology at the University of York under the supervision of Professor Ian Hitchcock and Dr David Kent. As the iCASE industrial partner, the CRUK/AstraZeneca Antibody Alliance Laboratory (AAL) will provide training and resources for your project, which will include at least two extended periods of research based at their laboratories in Cambridge.

At AAL, you will be trained in phage display to generate a broad panel of TPOR and EPOR-binding antibodies. You will then apply a range of high throughput screening biochemical and biophysical screening techniques (e.g. surface plasmon resonance (SPR) and Octet) to characterise the diverse binding characteristics and epitopes of your antibodies. At York, you will be trained to determine the functional and mechanistic effects of surrogate cytokines using cell lines and primary HSC assays, single molecule super resolution microscopy, dimerization reporter assays and high-throughput phospho-flow cytometry. 

For further information, please contact Professor Ian Hitchcock ([Email Address Removed]; @hitchcock_lab) or Dr David Kent ([Email Address Removed]; @scienceadvocacy).

Benefits of being in the DiMeN DTP:

This project is part of the Discovery Medicine North Doctoral Training Partnership (DiMeN DTP), a diverse community of PhD students across the North of England researching the major health problems facing the world today. Our partner institutions (Universities of Leeds, Liverpool, Newcastle, York and Sheffield) are internationally recognised as centres of research excellence and can offer you access to state-of the-art facilities to deliver high impact research.

We are very proud of our student-centred ethos and committed to supporting you throughout your PhD. As part of the DTP, we offer bespoke training in key skills sought after in early career researchers, as well as opportunities to broaden your career horizons in a range of non-academic sectors.

Being funded by the MRC means you can access additional funding for research placements, international training opportunities or internships in science policy, science communication and beyond. See how our current DiMeN students have benefited from this funding here: http://www.dimen.org.uk/overview/student-profiles/flexible-supplement-awards

Further information on the programme and how to apply can be found on our website:

http://www.dimen.org.uk/how-to-apply/application-overview