Umbilical cord blood T cells can be isolated and enriched by CD62L selection for use in 'off the shelf' chimeric antigen receptor T-cell therapies to widen transplant options
Christos Georgiadis,
Lauren Nickolay,
Farhatullah Syed,
Hong Zhan,
Soragia Athina Gkazi,
Annie Etuk,
Ulrike Abramowski-Mock,
Roland Preece,
Piotr Cuber,
Stuart Adams,
Giorgio Ottaviano,
Waseem Qasim
Affiliations
Christos Georgiadis
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London
Lauren Nickolay
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London
Farhatullah Syed
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London
Hong Zhan
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London
Soragia Athina Gkazi
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London
Annie Etuk
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London
Ulrike Abramowski-Mock
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London
Roland Preece
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London
Piotr Cuber
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London
Stuart Adams
Great Ormond Street Hospital for Children NHS Trust, WC1N 3JH, London
Giorgio Ottaviano
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London
Waseem Qasim
UCL Great Ormond Street Institute of Child Health, WC1N 1DZ, London, UK; Great Ormond Street Hospital for Children NHS Trust, WC1N 3JH, London
Umbilical cord blood (UCB) T cells exhibit distinct naive ontogenetic profiles and may be an attractive source of starting cells for the production of chimeric antigen receptor (CAR) T cells. Pre-selection of UCB-T cells on the basis of CD62L expression was investigated as part of a machine-based manufacturing process, incorporating lentiviral transduction, CRISPR-Cas9 editing, T-cell expansion and depletion of residual TCReeeT cells. This provided stringent mitigation against the risk of graft versus host disease (GVHD), and was combined with simultaneous knockout of CD52 to enable persistence of edited T cells in combination with preparative lymphodepletion using Alemtuzumab. Under compliant manufacturing conditions, two cell banks were generated with high levels of CAR19 expression and minimal carriage of TCReeeT cells. Sufficient cells were cryopreserved in dose-banded aliquots at the end of each campaign to treat dozens of potential recipients. Molecular characterisation captured vector integration sites and CRISPR editing signatures and functional studies, including in vivo potency studies in humanised mice, confirmed antileukaemic activity comparable to peripheral blood-derived universal CAR19 T cells. Machine manufactured UCB derived T cells banks offer an alternative to autologous cell therapies and could help widen access to CAR T cells.
