Tracking Metabolic Compartmentation And Epigenetic Metabolism In Human Lung Cancer.

Although metabolic reprogramming has been recognized as a hallmark of human cancers, our understanding of metabolic heterogeneities/compartmentation and regulation of epigenetics in human tumors is largely unknown. This is crucial to elucidating metabolic checkpoints and strategizing therapeutic target(s) for human cancer. The slow progress is at least in part due to the lack of relevant models and tools for studying these processes. We have established patient-derived organotypic tissue cultures (OTC) that recapitulate the in vivo metabolic reprogramming of human cancer while retaining native patient tumor microenvironment and 3D architecture 1,2. The matched pairs of cancerous (CA) and non-cancerous (NC) lung OTC from the same patient enable reprogrammed cancer tissue metabolism and on-/off-target drug responses for individual patients to be defined. We have deployed this system along with multiplexed Stable Isotope-Resolved Metabolomics (mSIRM) to probe the efficiency of different substates that fuel de novo synthesis of purine nucleotides in matched patients’ CA versus NC lung OTC 3. We found for seven non-small cell lung cancer (NSCLC) patients that 13C6-glucose was the preferred substrate over D3-serine, D2-glycine, or 13C2-glycine in supporting purine synthesis in CA tissues but not or less so in matched NC tissues. Kinetic modeling of the tracer data along with gene/protein expression data suggests that cytoplasmic activation and dynamic compartmentation of the glucose-to-serine pathway as well as reversal of mitochondrial serine to glycine fluxes can account for this preference for glucose. In all seven patients’ CA tissues, c-MYC was overexpressed and its suppression in NSCLC PC9 cells blocked de novo serine synthesis from glucose, enhanced serine-glycine exchanges, and reduced the efficiency of glucose versus serine incorporation into ATP/GTP. However, serine rather than glucose was the preferred substrate for purine synthesis in NSCLC cell lines but c-MYC could alter this preference for serine towards glucose in fueling purine synthesis. This tissue and cell line difference should be considered in designing serine starvation-based cancer therapy. To track metabolism that modulates epigenetic regulation, we employed a fast, sensitive, and robust Fourier transform-mass spectrometric method to analyze S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) levels in A549 cells under anti-cancer methylseleninic acid (MSA) and selenite treatments. We found that SAM/SAH levels were reduced by both treatments and MSA but not selenite increased the SAM to SAH ratio. These changes were accompanied by reduced proliferation, central metabolism, and H3K27Me3 levels in MSA and less so in selenite-treated A549 cells. Thus, the tool for tracking SAM/SAH metabolism along with the SIRM approach opens the opportunity for interrogating epigenetic metabolism and its regulation of growth-promoting central metabolic events. Funding: This work was supported in part by grants: 1R01ES022191-01 (to TWMF and RMH), 1P01CA163223-01A1 (to ANL and TWMF), and 1U24DK097215-01A1 (to RMH, TWMF, and ANL). Refs: 1 Sellers, K. et al. Pyruvate carboxylase is critical for non–small-cell lung cancer proliferation. The Journal of Clinical Investigation 125, 687-698, doi:10.1172/JCI72873 (2015). 2 Fan, T. W., Lane, A. N. \u0026 Higashi, R. M. Stable Isotope Resolved Metabolomics Studies in ex vivo Tissue Slices. Bio-protocol 6, e1730 (2016). 3 Fan, T. W. M. et al. De novo synthesis of serine and glycine fuels purine nucleotide biosynthesis in human lung cancer tissues. The Journal of biological chemistry 294, 13464-13477, doi:10.1074/jbc.RA119.008743 (2019). Citation Format: Teresa W-M Fan, Andrew N. Lane, Jason A. Brandon, Joonseon Yang, Chi V. Dang, Richard M. Higashi. Tracking metabolic compartmentation and epigenetic metabolism in human lung cancer [abstract]. In: Abstracts: AACR Special Virtual Conference on Epigenetics and Metabolism; October 15-16, 2020; 2020 Oct 15-16. Philadelphia (PA): AACR; Cancer Res 2020;80(23 Suppl):Abstract nr IA22.