Cancer cells have been known for some time to have very different metabolismas compared to that of normal non proliferating cells. As metabolism is involvedin almost every aspect of cell function, there has been a recent resurgence ofinterest in inhibiting cancer metabolism as a therapeutic strategy. Inhibitors thatspecifically target altered metabolic components in cancer cells are being developedas antiproliferative agents. However, many such inhibitors have not progressedinto the clinic due to limited efficacy either in vitro or in vivo. In this study weexplore the hypothesis that this is often due to the robustness of the metabolicnetwork and the differences between individual cancer cell lines in their metaboliccharacteristics. We take a systems biology approach. We investigate the cellular bioenergetic profiles of a panel of five non-small celllung cancer cell lines before and after treatment with a novel inhibitor of theglutaminase-1 (GLS1) enzyme. Additionally, we explore the effects of this inhibitoron intracellular metabolism of these cell lines as well as on the uptake and secretionof glucose, lactate and amino acids. To be able to do the latter robustly, wehad to modify the experimental assay considerably from procedures that seemto be standard in the literature; using these earlier procedures the metabolicenvironment of the cells was highly variable, leading to misleading results onthe metabolic effects of the inhibitor. We reduced cell density, altered mediumvolume and changed the time-window of the assay. This led to the cells growingexponentially, appearing indifferent to the few remaining changes. In this newassay, the metabolic effects of the glutaminase inhibitor became robust. One of the most significant results of this study is the metabolic heterogeneitydisplayed across the cell line panel under basal conditions. Differences in themetabolic functioning of the cell lines were observed in terms of both theirbioenergetic and metabolic profile. The amount of respiration attributed tooxidative phosphorylation differed between cell lines and respiratory capacity wasattenuated in most cells. However, the rate of glycolysis was similar betweencell lines in this assay. These results suggest that the Warburg effect arisesthrough a greater diversity of mechanisms than traditionally assumed, involvingvarious combinations of changes in the expression of glycolytic and mitochondrialmetabolic enzymes. The effects of GLS1 inhibition on cellular bioenergetics and metabolism alsodiffered between cell lines, even between resistant cell lines, indicating that theremay also be a diversity of resistance mechanisms. The metabolomic response ofcell lines to treatment suggests potential resistance mechanisms through metabolicadaptation or through the prior differences in the metabolic function of resistantcell lines. Part of the metabolome response to GLS1 inhibition was quite specificfor sensitive cells, with high concentrations of IMP as the strongest marker. Our results suggest that the metabolome is a significant player in what determinesthe response of cells to metabolic inhibitors, that its responses differ between cancercells, that responses are not beyond systems understanding, and that thereforethe metabolome should be taken into account in the design of and therapy withanti-cancer drugs.

GE Healthcare Bio-Sciences AB in Umeå produce a variety of chromatography systems. One of the main components in chromatography is the UV module, which measure the light absorption of different wavelengths in the liquid being pumped through a cell. Currently at the Umeå site two types of UV-cells are produced; lab cells and industrial cells. The current test station for the industrial UV-cells is outdated, in disrepair and no longer supported.  GE has developed a test station for the lab cells that evaluates UV and flow properties, the data is stored in GE’s own production database, Prodas. The aim of this work is to design a test station for industrial UV-cells to improve the quality of the cells. The primary goal is a test station that can measure pressure, flow and absorption. The secondary goal is to discover and if possible, implement solutions that will streamline and automate the test station. A prototype of a test station for industrial UV-cells based on that for lab cells has been developed. The solution consists of an adapter that links the light path from the monitor through the UV-cell to the detector. The test station can measure pressure, flow and absorption but can only perform absorption and leakage tests.  Automation and efficiency have been accomplished in the form of scripts used to conduct absorption and leakage tests. The test station requires further development before it can be used in the production line. / GE Healthcare Bio-Sciences AB i Umeå tillverkar ett flertal system, ett av systemen är vätskekromatografen. Vätskekromatografen är en kemisk separationsmetod som använder sig av en UV-monitor, UV-detektor och en UV-cell för att mäta absorptionen av en lösning och framta koncentrationen av det eftersökta ämnet. På anläggningen i Umeå tillverkas två typer av UV-celler; laborationceller och industriceller. För labbcellerna har GE utvecklat en teststation som testar och utvärderar UV och flödesegenskaper samt lagrar data i GE:s egen produktionsdatabas, Prodas. Den befintliga stationen för industriceller är äldre och omodern, en utveckling behövs för att upprätthålla högre kvalité. Syftet med detta projekt är att uppdatera teststationen för industriella UV-celler till samma nivå som stationen för laborationsceller. Det primära målet är att konstruera och designa en teststation för industriella UV-celler som kan mäta tryck, flöde och absorption. Det sekundära målet är att upptäcka och om möjligt verkställa lösningar som kommer effektivisera och automatisera mätningarna. En prototyp av en teststation för industriella UV-celler baserat på den för laborationceller har framtagits, och består av en adapterlösning som används för att sammanlänka ljusbanan från monitorn genom flödescellen till detektorn. Prototypen kan enbart utföra absorption och läckagemätningar. Automatisering och effektivisering har utförts i form av scripts som används för att genomföra absorptions och läckagemätning. Teststationen kräver fortsatt vidareutveckling innan den kan används i produktionslinjen.