Development of systems for the generation of red blood cells (RBCs) from stem cell sources (adult peripheral blood, cord blood and induced pluripotent stem cells) in vitro is of great interest to health services globally, as such a product could overcome many of the problems associated with donor blood, in particular the availability of certain rare blood groups and for patients requiring repeat transfusions.
However, adult and cord stem cells have finite expansion potential which presently restricts the number of red cells that can be obtained, greatly impacting the economic viability of producing therapeutic quantities of red cells from these sources. In addition iPSC-derived erythroid cells have severe differentiation defects. We have therefore taken an alternative approach creating immortalized adult erythroid cell lines that recapitulate erythropoiesis, and which provide the first viable approach for a sustainable source of RBCs for therapeutics. Moreover we have used CRISPR gene editing to create designer lines with specific blood group phenotypes and as a universal RBC product.
However, such lines have significant additional potential, and we are using our methodology and gene editing to create lines as disease model systems to study the underlying molecular defects of conditions such as thalassemia, sickle cell disease and congenital dyserythropoietic anaemias, along with the application of these lines as novel drug screening platforms.
We are also interested in the regulation of erythropoiesis, in particular transcription factors such as KLF1, which is essential for erythropoiesis with mutations resulting in severe red blood cell disorders.
We utilise cell biology, transcriptomic, proteomic, biochemical and molecular biology approaches, including gene editing by CRISPR, shRNAs and ectopic expression, qPCR, multiplex flow cytometry, Fluorescent activated cell sorting, confocal microscopy, TMT labelling coupled Mass Spec.
We are part of the NIHR Blood and Transplant Research Unit (BTRU) to advance research on the manufacture of red blood cells from stem cells and their translation from the lab to human trials, along with researchers at NHS Blood and Transplant (NHSBT, Bristol) and Dr Ash Toye within the University.
Available projects include:
· Creation of cytokine independent erythroid lines to aid development of therapeutic products and to further investigate the regulation of erythropoiesis
· Creation and characterisation of specific red blood cell disease lines to investigate the molecular basis of disease and facilitate development of novel therapeutic approaches
· Determining how KLF1 mutations result in both clinically benign and severe disease phenotypes
· Genome-wide approach to identify novel genes as therapeutic targets for β-thalassemia via CRISPR library screening
· Identification of novel targets for treatment of β-haemoglobinopathies via targeted kinase inhibitor screening