Abstract B179: 2-Deoxy-D-glucose (2-DG) combination with ketogenic diet induces sedation and hypothermia in mice that becomes lethal

Abstract

Abstract Introduction. The term “ketogenic” was coined for the diet that specifically consists of a 4:1 ratio of fats to proteins. 2-Deoxy-d-glucose (2-DG) is a synthetic glucose analog that potently inhibits glycolysis. It has been hypothesized that 2-DG in combination with KD, should improve the anticancer effects of either therapeutic approach. The therapeutic benefit of such a combination was supported by studies of Voss et al. (Int J Mol Sci, 2018. 19(8): PMC6121440), showing that continued combination of KD and anti-glycolytic therapy is feasible, and KD increased tolerance to 2-DG. However, in contrast to this report, our preliminary studies revealed a dose-dependent reduction in the survival of mice of receiving inhibition of glycolysis (2-DG) and KD. The objective of this study was to assess the toxic limitation of the combination of glycolysis inhibition and ketogenic diet. Methods. CD-1 mice were maintained on either a standard diet (SD) or a ketogenic diet (KD). Some groups were treated with of 2-DG at 1.5 g/kg via the oral or intraperitoneal route or vehicle. For temperature monitoring, the animals were implanted subcutaneously with a temperature microchip UCT-2112. Body temperature was recorded manually for each mouse using a contactless, handheld URH-1HP receiver. Alternatively, for continuous monitoring of the body temperature, the UID Mouse Matrix was used. Mice exhibiting treatment-related toxicity symptoms, such as prolonged hypothermia or laborious breathing, and euthanized following blood collection for hematology and serum chemistry analysis. Key organs were also collected for histopathology analysis. Results. Previously, we reported that 2-DG reduces the survival of mice maintained on KD (median survival 9 days p=0.0022), while it is well tolerated by mice receiving regular chow (no deaths reported). Here, we report that the administration of 2-DG induces transient sedation and hypothermia. CBC analysis revealed that the number of platelets in the KD + 2-DG group was significantly decreased compared to the SD + 2-DG group and the KD + vehicle group (p<0.001). The reduction of platelets was associated with a two- to three-fold increase of LDH in the KD + 2-DG group. Histopathological analysis of the lung tissue revealed the presence of diffuse congestion in blood vessels and dilation of the alveolar capillaries consistent with interstitial pneumonia. Similarly, degenerative and congestive hepatic parenchyma was observed in animals treated with 2-DG and KD. Preliminary experiments involving supplementation of 2-DG-treated animals with D-glucose or D-mannose failed to provide a rescue effect. Conclusion. A highly reproducible, dose-dependent toxicity leading to rapid death of treated animals maintained on KD was observed following 2-DG administration, while the same doses appeared to be safe in mice fed a standard diet. The toxic effects induced by the combination treatment included acute thrombocytopenia and acute interstitial pneumonia; however, delineation of the exact mechanism of toxicity requires additional studies. Citation Format: Rafal Zielinski, Moises Martinez Castillo, Damian Grybowski, Krzysztof Grela, Roberto Cardenas-Zuniga, Izabela Fokt, Stanislaw Skora, Gerardo Ramirez, Waldemar Priebe. 2-Deoxy-D-glucose (2-DG) combination with ketogenic diet induces sedation and hypothermia in mice that becomes lethal [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr B179.