Abstract 1035: Targeting glutamine metabolism with the novel inhibitor JHU-083 inhibits tumor growth and alters the tumor immune microenvironment
Authors: Englert Judson , Cheng Chih-Hsien , Leone Robert , Majer Pavel , Rais Rana , Slusher Barbara , Powell Jonathan 
Published: July 2016

Glutamine metabolism plays a critical role in promoting multiple biosynthetic pathways necessary for tumor growth. Therefore we endeavored to create a small molecule inhibitor that would block multiple pathways of glutamine metabolism as opposed to blocking one enzyme. Previous studies employing the non-specific glutamine antagonist 6-diazo-5-oxo-L-norlecuine (DON) have achieved this goal however, with much toxicity. To this end we devised a novel prodrug of DON (JHU-083) with distinct distribution and cellular partitioning properties. When dosed on an equimolar basis, JHU-083 retains the antitumor efficacy of DON but is devoid of the gut toxicity and morbidity/mortality.

To this end Wild-type mice were injected subcutaneously EL4 T-cell lymphoma cells. The tumors were allowed to engraft and treatment was initiated on day 5 with either vehicle, DON or JHU-083. After 7 days of treatment there was complete tumor regression in both the DON and JHU-083 treated mice, while tumors continued to grow in the vehicle treated mice. Importantly, after 7 days of treatment the DON treated mice appeared ill with lethargy and ruffled fur and even death as a result of treatment toxicity. Alternatively, the JHU-083 treated mice remained healthy and vibrant.

Based on these studies in a lymphoma model we next tested JHU-083 in 2 models of colon cancer. To this end we injected wild-type mice subcutaneously with MC38 or CT26 cells. Again, similar to the EL4 lymphoma experiments, we observed that JHU-083 resulted in complete regression of MC38 and CT26 tumors after 4 and 7 days of treatment respectively.

The tumor microenvironment, which promotes immune evasion, is very much influenced by tumor metabolism. Thus, we wanted to determine the effect of inhibiting glutamine metabolism on the tumor immune microenvironment. To this end mice injected with B16 melanoma cells were treated for 3 days with DON during which time the tumors decreased in size. Interestingly, there was also a concomitant decrease in the percentage of Foxp3+ cells and an increase in the CD8:Foxp3+ T cell ratio amongst the TILS.

Overall our studies demonstrate the principle of cellular selectivity based upon demand. That is, a non-selective inhibitor of glutamine metabolism can robustly inhibit tumor growth while demonstrating minimal side effects based on the differential demand of cancer cells versus normal cells. Furthermore, our studies suggest that sequencing anti-metabolic therapy with immunotherapy might prove to be a novel means of broadening and enhancing the role of immunotherapy for the treatment of cancer.

Citation Format: Judson Englert, Chih-Hsien Cheng, Robert Leone, Pavel Majer, Rana Rais, Barbara Slusher, Jonathan Powell. Targeting glutamine metabolism with the novel inhibitor JHU-083 inhibits tumor growth and alters the tumor immune microenvironment. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1035.