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Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide. The expression of glucose transporter isoform 1, a key factor in transporting glucose into cancer cells, is overexpressed in several human cancers, including HCC. In addition, this has been shown to correlate with a higher proliferation index and more advanced stages in HCC, suggesting that inhibition of glucose metabolism is a promising therapeutic strategy. Our study used high-content screening (HCS) for compounds that target glucose metabolism and effect cell death in HCC cells. Specifically, we showed that a fluorescent 2-deoxyglucose analog, 2-[N-(7-nitrobenz-2- oxa-1,3-diazol-4-yl)amino]-2-deoxyglucose, and CellTrace Calcein Red-Orange AM can be used reliably as readouts for glucose uptake and proliferative index, respectively, to identify drug candidates that simultaneously reduce glucose uptake and induce cell death in HCC cells. Thus, fluorescent glucose uptake bioprobes can be implemented in HCS assays to identify previously unknown regulators of glucose metabolism in HCC. In addition, our study also employs the use of feedback system control (FSC.II), a platform that optimizes the combinations of drugs identified through HCS. The coordinated use of HCS and FSC.II can improve the development of drug combinations and uncover previously unidentified signaling pathways that govern HCC as well as other cancers.

Identification and Optimization of Combinatorial Glucose Metabolism Inhibitors in Hepatocellular Carcinomas