There and back again - reprogramming gamma/delta T-cells (gdT) into iPSCs in order to differentiate them back into (gdT) for cancer immunotherapy

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Bioprocessing of human T-cell products have begun to revolutionize cancer treatments, in particular in the fields of chimeric antigen receptor (CAR) use in leukaemias and beyond. In this project we aim to build on our respective strengths in the UCL Biochemical Engineering dept. (e.g. 1, 2) and the Steve Oh group at the Bioprocessing Technology Institute (BTI) in Singapore (e.g. 3, 4) - by investigating a novel and potentially more useful way to produce genetically modified and target-specific gdT cells for use in immunotherapies in the future. BTI has developed patented technologies in 1) reprogramming T cells into hiPSC and 2) directed differentiation into haemapoietic stem cells (HSC). HSC can be further differentiated to T cells by a combination of growth factors or activation of transcription factors such as Foxp3. BTI has demonstrated the ability to differentiate hiPSC to retinal pigment epithelial cells (RPE) by activation of Pax6 transcription factor. This technology has just been filed as a new platform patent. A similar approach is likely to work for accelerating T cell differentiation.The student will aim to do this by culturing gdT cells from peripheral blood mononuclear cells and other epithelial sources. He will then derive hiPSCs from the gdT using established procedures (5, 6) These hiPSCs will then be maintained and passaged as stem cells (SC) and characterised in detail. The student will then (re)differentiate the gdT-derived hiPSCs back into HSC, essentially as previously described (5, 6). From these HSC she will subsequently attempt to derive fully differentiated gdT. As part of this project, we will importantly be able to genetically engineer the gdT in a convenient way during the SC stage - which we hypothesise will lead to retention of the engineered gene and its expression. This could involve a TCR, co-stimulatory molecules, chimeric antigen receptors (CARs), or 'knock-out' of genes such as MHC (HLA).1. Barisa M. et al., Sci Rep. 2017, 7(1): 28052. Fisher J. et al., Mol Ther. 2017, 26(2): 3543. Lee et al., Biotechnol J. 2018, 13(4): e17005674. Sivalingam J. et al., Haematologica 2018, 103(7): e2795. Watanabe D. et al., Stem Cells Transl Med. 2018, 7(1): 346. Zeng J. et al., PLoS One. 2019, 14(5): e0216815