Abstract 2537: Beta-glucan reprograms immunomodulatory metabolism in human macrophage and ex vivo in lung cancer tissues

Abstract We combined multiplex Stable Isotope-Resolved Metabolomics (mSIRM) with Reverse Phase Protein Array (RPPA) to map the time course changes of IM metabolic network and key protein regulators in four human donors’ MΦ’s in response to differential polarization and whole glucan particulates (WGP) treatments. We found consistent and plastic network responses to polarization durations and WGP treatments compared to those of the mouse counterparts. Consistent responses included enhanced 15N2-tryptophan catabolism to quinolinate, 2H2-glucose oxidation to ribose/ribulose-5-phosphate, and conversion of 13C5-glutamine to itaconate in response to pro-inflammatory (M1) versus anti-inflammatory (M2) stimuli. WGP robustly induced the buildup of 2H2-glucose-derived 6-phosphogluconate, -lactate, and -IMP, 13C5-glutamine-derived fructose-1,6-bisphosphate, and increased enrichment of 2H-labeled UDP-N-acetylglucosamine in M2-MΦ’s. However, the Krebs cycle activity was variably enhanced by M2 stimuli or WGP treatment. The consistent effects were related to increased release of proinflammatory IM effectors IL-1β/CXCL10/IFNγ/TNFα by M1-MΦ’s and enhanced release of IL-1β/TNFα to above M1-MΦ’s levels in WGP-treated M2-MΦ’s while boosting the latter in anti-inflammatory IL-10 release and maintenance of NAD+ synthesis. They were also related to lower phagocytosis in M1-MΦ’s and WGP-treated M2-MΦ’s versus M2-MΦ’s. Together with the expression changes of key protein regulators, we suggest enhanced tryptophan catabolism with blocked NAD+ and UTP synthesis to be key to the consistent changes in immune functions in response to M1 stimuli. Likewise, increased glucose utilization via glycolysis and the oxidative branch of the pentose phosphate pathway, and blockade of glutamine-fueled N-linked glycosylation could be linked to reversion of M2 to M1-type immune functions. Reprogrammed Krebs cycle and glutamine conversion to UTP occurred variably in WGP-treated ex vivo organotypic tissue cultures (OTCs) of human non-small cell lung cancer (NSCLC), which could reflect variable M1 repolarization of tumor associated MΦ’s. This in turn correlated with IL-1β/TNFα releases and compromised tumor status, making patient-derived OTCs a unique model for studying variable immunotherapeutic efficacy in cancer patients. In conclusion, consistent and variable IM metabolic responses were evident in four human donors’ MΦ’s. WGP repolarized some M2 to M1-type responses while boosting other M2-type responses. NSCLC OTCs from six patients showed variable M1 repolarization in response to WGP. Citation Format: Teresa W. M. Fan, Saeed Daneshmandi, Teresa A. Cassel, Mohammad B. Uddin, James Sledziona, Patrick T. Thompson, Penghui Lin, Richard M. Higashi, Andrew N. Lane. Beta-glucan reprograms immunomodulatory metabolism in human macrophage and ex vivo in lung cancer tissues [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2537.