Enhancing human NK cell development: from UCB-CD34+ cells to functional NK cells
File(s)
Author(s)
Ullmo, Ines
Type
Thesis
Abstract
Natural Killer (NK) cells are innate lymphocytes that have shown great promise for use in cancer immunotherapy. In comparison to T cells, NK cells can mount quicker immune responses and recognise a wider variety of tumour-associated antigens. Moreover, approved T cell-based immunotherapies are autologous, which can lead to manufacturing inefficiencies and low accessibility of such therapies. In contrast, NK cell therapies can be allogeneic without causing adverse reactions or rejection, thereby allowing for off-the-shelf NK cell therapies to be manufactured from universal donors and used on demand. One of the main limitations for the development of NK cell immunotherapies is the production and expansion of sufficient NK cells ex vivo to meet the therapeutic cell requirement. In this thesis, I optimised a robust in vitro method for NK cell development from umbilical cord blood (UCB) CD34+ cells that generates large numbers of cytotoxic NK cells. Moreover, this optimised method of in vitro NK cell development led to the production of CD56+CD16+ NK cells, which have the potential to exhibit ADCC when encountering opsonised target cells. The generation of CD56+CD16+ NK cells from UCB CD34+ cells was achieved by expanding CD34+ cells in pre-differentiation media (4-6 days) prior to start of NK cell development. Moreover, activating Notch1 receptor signalling (through DLL4) prior to NK cell development further increased total NK cell production, including CD56+CD16+ NK cells. Adding Remodelin, a Nat10 specific chemical inhibitor, to CD34+ cells prior to start of in vitro NK cell development also slightly enhanced NK cell production under specific conditions. To investigate the molecular mechanisms responsible for these processes that enhance NK cell development, a gene editing method was put in place and thoroughly optimised, to correlate genotype, phenotype and function of knockout CD34-derived NK cells. Therefore, this thesis contributes to developing more efficient NK cell-based immunotherapies.
Version
Open Access
Date Issued
2023-07-30
Date Awarded
2024-06-01
Copyright Statement
Attribution-NonCommercial 4.0 International Licence (CC BY-NC)
License URL
Advisor
Brady, Hugh
Sponsor
Medical Research Council (Great Britain)
Publisher Department
Life Sciences
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)