Monitoring the metabolic fate of aspartic acid in liver cancer through NMR spectroscopy

Liver cancer, more specifically hepatocellular carcinoma (HCC) is one of the most common types of cancer worldwide. There is an unmet and urgent clinical need not only for more effective and targeted therapeutics but also for clear-cut and better diagnostic tools for early detection and accurate assessment of liver cancer. Cell culture of HCC cells (Huh-7 cell line) was done using DMEM with 10% FBS and doped with 5 mM 13C-aspartic acid for 1hr and 48 hrs. A control without aspartic acid was also prepared and tested. The samples were done in triplicate and measured using a 600 MHz NMR spectrometer. Both cell and media extracts were analyzed to obtain NMR results. Preliminary NMR results show that aspartic acid entered the tricarboxylic acid (TCA) cycle through oxaloacetate and produced a number of TCA cycle metabolites. This study is supported by the Welch Foundation grant AT?2111?20220331, US Department of Defense CDMRP grants W81XWH-21-1-0176, W81XWH-22-1-0105, W81XWH-19-1-0741, and W81XWH-22-1-0003.