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

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): 2805
2. Fisher J. et al., Mol Ther. 2017, 26(2): 354
3. Lee et al., Biotechnol J. 2018, 13(4): e1700567
4. Sivalingam J. et al., Haematologica 2018, 103(7): e279
5. Watanabe D. et al., Stem Cells Transl Med. 2018, 7(1): 34
6. Zeng J. et al., PLoS One. 2019, 14(5): e0216815

The CDT has a proven track record of delivering impact from its research and training activities and this will continue in the new Centre. The main types of impact relate to: (i) provision of highly skilled EngD and sPhD graduates; (ii) generation of intellectual property (IP) in support of collaborating companies or for spin-out company creation; (iii) knowledge exchange to the wider bioprocess-using industries; (iv) benefits to patients in terms of new and more cost effective medicines, and (v) benefits to the wider society via involvement in public engagement activities and impacts on policy.

With regard to training, provision of future bioindustry leaders is the primary output of the CDT and some 96% of previous EngD graduates have progressed to relevant bioindustry careers. These highly skilled individuals help catalyse private sector innovation and biomanufacturing activity. This is of enormous importance to capitalise on emerging markets, such as Advanced Therapy Medicinal Products (ATMPs), and to create new jobs and a skilled labour force to underpin economic growth. The CDT will deliver new, flexible on-line training modules on complex biological products manufacture that will be made available to the wider bioprocessing community. It will also provide researchers with opportunities for international company placements and cross-cohort training between UCL and SSPC via a new annual Summer School and Conference.

In terms of IP generation, each industry-collaborative EngD project will have direct impact on the industry sponsor in terms of new technology generation and improvements to existing processes or procedures. Where substantial IP is generated in EngD or sPhD programmes, this has the potential to lead to spin-out company creation and job creation with wider economic benefit. CDT research has already led to creation of a number of successful spin-out companies and licensing agreements. Once arising IP is protected the existing UCL and NIBRT post-experience training programmes provide opportunities for wider industrial dissemination and impact of CDT research and training materials.

CDT projects will address production of new ATMPs or improvements to the manufacture of the next generation of complex biological products that will directly benefit healthcare providers and patients. Examples arising from previous EngD projects have included engineered enzymes for greener pharmaceutical synthesis, novel bioprocess operations to reduce biopharmaceutical manufacturing costs and the translation of early stem cell therapies into clinical trials. In each case the individual researchers have been important champions of knowledge exchange to their collaborating companies.

Finally, in terms of wider public engagement and society, the CDT has achieved substantial impact via involvement of staff and researchers in activities with schools (e.g. STEMnet), presentations at science fairs (Big Bang, Cheltenham), delivery of high profile public lectures (Wellcome Trust, Royal Institution) as well as TV and radio presentations. The next generation of CDT researchers will receive new training on the principles of Responsible Innovation (RI) that will be embedded in their research and help inform their public engagement activities and impact on policy.