Metabolic targeting of cancer cells via the glyoxalase detoxification system

Cancer metabolism is an area of research that is experiencing a renaissance as new models and methodologies add to Otto Warburg?s pioneering work from nearly 80 years ago. Warburg found that tumors are often ?addicted? to glucose even when grown in the presence of oxygen. Aerobic glycolysis or the Warburg effect continues to be used in the clinic today to image a variety of tumor types using 2-deoxy-2-[18F]-D-glucose positron emission tomography (18FDG-PET). Sustained elevation of glycolysis can lead to the accumulation of an endogenous, toxic metabolite called methylglyoxal (MG). MG is an alpha-oxoaldehyde and results from the breakdown of glucose and the dephosphorylation of triose phosphates that are key intermediates of glycolysis. MG is highly reactive with cellular nucleophiles and can react with cellular biopolymers such proteins, DNA, RNA and lipids, resulting in the formation of advanced glycation end-products (AGEs). The tumor cell combats this by the detoxification of MG using the highly conserved, glutathione-dependent glyoxalase I/II (Glo I/II) pathway. We hypothesize that the Glo I/II pathway may represent a target in difficult to treat tumor types that are glycolytic. Specific Aims include: [1] To establish a panel of cell lines (triple negative breast cancer and castration-resistant prostate cancer) that are appropriate for this work and to train Master?s and undergraduate students in proper aseptic technique. [2] To determine the proof of principle of Glo I/II as a target using pharmacologic and molecular biological approaches to inhibit or knock down Glo I/II. These discrete aims will be used to build a long-term research project that I will seek NIH SCORE funding for. Long-term, the effect of Glo I/II inhibition will be further explored by interrogating compensatory responses by the aldo-keto reductase 1B1 and 1B10 (AKR1B1, 1B10) which is also known to reduce MG. Long-term, we will also seek to quantify intracellular MG using gas chromatography mass spectrometry (GC/MS) and use liquid chromatography tandem mass spectrometry (LC/MS/MS) to quantify the DNA-AGE, N2-(1-carboxyethyl)-2?-deoxyguanosine (CEdG) (in collaboration with my former mentor/collaborator at the Beckman Research Institute, City of Hope).