Quantifying metabolic reprogramming in the context of Natural Killer cell immunotherapy against Hepatocellular carcinoma using conventional and microscale NMR spectroscopy

Abstract

Metabolic reprogramming is at the core of cell functionality and fate. The ability to alter the use of metabolic pathways is observed both in the immune system and as a hallmark of cancer. This thesis demonstrates the use of nuclear magnetic resonance (NMR) spectroscopy to quantify the aberrant metabolism of hepatocellular carcinoma (HCC) and the metabolic reprogramming of natural killer (NK) cells. NK cell immunotherapies rely on activation by cytokines but can be hampered by the immunosuppresive environment of solid tumours such as HCC. Quantification of this effect is important in the development of immunotherapies against solid cancers as well as for understanding disease mechanisms. NK cells are known to undergo significant metabolic reprogramming in response to external stimuli in order to increase cytotoxic function. A similar phenomenon is observed in the aberrant metabolism of cancer cells that allows the utilisation of diverse sources of energy to support rapid growth and proliferation. NMR spectroscopy is a powerful technique that can be utilised in metabolomics for non-invasive quantification of multiple metabolites simultaneously from complex mixtures such as biological samples. This thesis demonstrates the power of NMR for non-invasive quantification of metabolic profiles, both in parallel with conventional cell culture methods and combination with microfluidic in operando culture of HCC and NK cells. This research explores the applications of both conventional and microfluidic NMR in investigating the metabolic reprogramming undergone by cells during tumourigenic transformation and immune responses. The combination of NMR and a novel microfluidic lab-on-a-chip device is demonstrated, enabling in situ culture of primary immune cells and HCC cell lines. The μNMR system includes a hydrothermal heating system to enable continuous in situ NMR observation of cell culture over a 24 hour period and is able to measure millimolar concentrations at microlitre volumes.

More information

Identifiers

ORCID for Salim Khakoo: orcid.org/0000-0002-4057-9091

ORCID for Marcel Utz: orcid.org/0000-0003-2274-9672

Catalogue record

Export record