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

人类t细胞产品的生物加工已经开始彻底改变癌症治疗，特别是在白血病和其他疾病的嵌合抗原受体（CAR）使用领域。在这个项目中，我们的目标是建立我们各自在UCL生物化学工程系（例如1,2）和新加坡生物处理技术研究所（BTI）的Steve Oh小组（例如3,4）的优势-通过研究一种新的和可能更有用的方法来生产转基因和靶向特异性gdT细胞，用于未来的免疫治疗。BTI已经开发了专利技术，1)将T细胞重编程为hiPSC， 2)定向分化为造血干细胞（HSC）。HSC可以通过结合生长因子或激活转录因子如Foxp3进一步分化为T细胞。BTI已经证明了通过激活Pax6转录因子将hiPSC分化为视网膜色素上皮细胞（RPE）的能力。这项技术刚刚被申请为新的平台专利。类似的方法可能对加速T细胞分化起作用。学生将通过培养外周血单核细胞和其他上皮细胞来源的gdT细胞来实现这一目标。然后，他将使用既定的程序从gdT中提取hiPSCs(5,6)，然后将这些hiPSCs作为干细胞（SC）进行维持和传代，并详细表征。然后，学生将（重新）将gdt衍生的hiPSCs分化回HSC，本质上如前所述（5,6）。随后，她将尝试从这些HSC中推导出完全分化的gdT。作为该项目的一部分，我们将能够在SC阶段以一种方便的方式对gdT进行基因工程，我们假设这将导致工程基因及其表达的保留。这可能涉及TCR、共刺激分子、嵌合抗原受体（CARs）或MHC (HLA) 1等基因的“敲除”。李志强，刘志强，刘志强，等。中国生物医学工程学报，2017,32(1):444 - 444。李海峰，等。生物化学学报，2017,26(2):3543。李建军，李建军，李建军，等。中国生物医学工程学报，2018,13(4):771 - 774。李志强，王志强，等。中华血液学杂志，2018,103(7):779 - 779。杨建军，张建军，张建军，等。中国生物医学工程学报，2018,31(1):346。[j] .科学通报，2019,14(5):0216815

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