Abstract PD4-11: [18F]Fluciclovine PET tracks cellular glutamine pool size in breast cancer and changes in response to metabolic inhibition

Abstract

Abstract Background: Some forms of triple negative breast cancer (TNBC) rely on glutamine (Gln) metabolism for survival and growth (1), therefore, targeting this metabolic pathway provides a viable strategy for managing TNBC. Drugs that inhibit glutaminase (GLS), a key enzyme of glutaminolysis, are being developed (1,2). [18F]Fluciclovine is a PET imaging agent that enters/exits cells via glutamine transporters and undergoes minimal metabolism. Therefore, we hypothesize, that akin to our prior work with [18F]fluoroglutamine (3), the distribution volume (VT) of fluciclovine obtained from dynamic PET can be used to estimate the cellular glutamine level (pool size) and to mark the effect of pharmacological inhibitors of tumor glutaminase (GLS). We tested this hypothesis in human TNBC and ER+ breast cancer xenograft exhibiting a high and low GLS activity, respectively. Methods: To make [18F]fluciclovine preparation suitable for mouse imaging, citrate in the formulation was removed and replaced with PBS by eluting through a column (Bio-Rad). Cellular uptake was performed in the presence and absence of Gln transporter inhibitors and GLS inhibitor. In vivo dynamic PET imaging were performed on mice bearing HCC1806 (TNBC) and MCF-7 (ER+ BC) xenografts. Dynamic PET images were analyzed by Logan Plot (PMOD) to estimate VT. Results: Cellular uptake of [18F]fluciclovine in HCC1806 and MCF-7 cells were sensitively inhibited by cold glutamine (Gln) and GPNA (a pharmacologic inhibitor of ASCT-2), confirming that the uptake is mediated by Gln transporters. The peak uptake in MCF-7 cells was 5-fold higher than HCC1806. In mouse models, VT from in vivo [18F]Fluciclovine PET in MCF-7 tumor is 1.4-fold of HCC1806. These data are consistent with a higher cellular Gln pool size in MCF-7 as the result of its lower GLS activity. After inhibition of tumor GLS activity, VT of [18F]fluciclovine in HCC-1806 tumors was increased by 56% from baseline values (n=2), whereas VT in MCF-7 tumors decreased 1% after treatment (n=2). Only a small change of FDG PET signal (5% decrease, n=5) was detected in TNBC tumors after GLS inhibitor treatment. Discussions: These data suggest that VT obtained from [18F]fluciclovine PET is sensitive to changes of the Gln pool size induced by GLS inhibition whereas FDG PET is not. Since the Gln pool size is inversely related to the GLS activity, increased VT is consistent with the increased intracellular Gln level when metabolic conversion of Gln to glutamate by GLS is inhibited. Our results suggest that [18F]fluciclovine, an imaging agent approved for prostate cancer imaging, may be useful for assessing glutamine pool size in breast cancer and changes in response to GLS inhibition Support: R21CA198563, R01CA211337, and Komen SAC130060. We thank Blue Earth Diagnostics for supplies of [18F]fluciclovine. 1.Gross MI, Demo SD, Dennison JB, et al. Mol Cancer Ther 2014;13(4):890-901. 2.Le A, Lane AN, Hamaker M, et al. Cell Metab 2012;15(1):110-21. 3.Zhou R, Pantel AR, Li S, et al. Cancer Res 2017;77(6):1476-84. Citation Format: Cao J, Choi H, Pantel A, Kranseler D, Lee H, Mankoff D, Zhou R. [18F]Fluciclovine PET tracks cellular glutamine pool size in breast cancer and changes in response to metabolic inhibition [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr PD4-11.