Metabolic Profiling of Multiple Myeloma Cancer Cells During Exposure to Chemotherapeutic Drugs: A Study of the Effects of ATX-101
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This thesis investigates the metabolomic stress response induced in JJN-3 cells treated with the cell-penetrating peptide ATX-A, the anti-cancer drug ATX-101, the chemotherapeutic drug cisplatin, and combinations of these drugs, along with untreated JJN-3 cells. This was achieved by using cell counts, glutamine and glucose uptake, and lactate excretion to calculate carbon flux, and by performing targeted metabolic profiling, with GC-QqQ-MS and Cap IC-MS/MS, and non-targeted metabolic profiling, with LC-QTOF-MS in both reverse phase and amide mode.Cell counting revealed that the anti-cancer drug ATX-101 has a growth inhibiting effect on JJN-3 cells, that some of this effect is caused by the cell penetrating effect of the drug and not the APIM sequence, and that there are biological variations between the cell suspensions grown in different cell culture flasks. The carbon flux results showed that JJN-3 cells use 40 80 % of the carbon from glutamine and glucose on lactate production, and 0.5 6 % on biomass production. They also indicated that the preferred carbon source for the JJN-3 cells is glucose, and that the cells treated with ATX-101 use less of the carbon for proliferation and more of the carbon for aerobic glycolysis than the remaining five treatments. Results from targeted and non-targeted metabolic profiling showed that the experiment design used in this thesis gives reproducible data despite of there being some biological variances between the three experiments performed, and that treatments performed on the JJN-3 cells have an immediate effect on the metabolome that should be investigated further. They also indicated that ATX-101 accounts for most of the effect seen at 4, 8, and 24 hours after addition of treatment, that the stress response induced by ATX-101 is caused by the cell penetrating peptide part of the drug as well as the APIM sequence, and that cisplatin has little or no effect on metabolome of JJN-3. Most of the variance in the results from the targeted metabolic profiling was caused by the metabolites glutamate, lactate, pyruvate, UDP-acetyl, and citrate.