Abstract 4230: Visualizing the distribution of metabolites and the efficacy of BIBF1120 on metabolic status of lung cancer derived tumors by imaging mass-spectrometry (MS)
Authors: Arai Daisuke , Yasuda Hiroyuki , Soejima Kenzo , Kagawa Shizuko , Hamamoto Junko , Nukaga Shigenari , Naoki Katsuhiko , Emoto Katsura , Sugiura Yuki , Suematsu Makoto , Betsuyaku Tomoko 
Published: July 2016

Abstract
Background: Imaging MS (Shimadzu CO., LTD.) is a novel technology that can quantitatively visualize the distribution of hundreds of metabolites in the selected areas of tissues. BIBF1120 (gift from Boehrlinger Ingelheim Seiyaku CO., LTD.) is a multiple tyrosine kinase inhibitor, targeting VEGFR, FGFR and PDGFR, and is known as an anti-angiogenic agent. Anti-angiogenic drugs are supposed to induce under-nutrition in tumors. However, no study has visualized the regional differences in the metabolic status of tumors induced by anti-angiogenic agents.
Aim: The aims of this study are to visualize the distribution of metabolites in tumors and metabolic status induced by BIBF1120 and to obtain the direct proof that BIB1120 induce under-nutrition status in tumors by Imaging MS.
Method: We used lung cancer derived cell lines, namely, PC9, H1975, H3122, A549, H69, and H520. We evaluated distribution of metabolites in tumors derived from xenograft models by Imaging MS. The efficacy of BIBF1120 was evaluated by MTS assay and mouse xenograft model. We treated the engrafted mice with BIBF1120 or vehicle for 4 weeks and harvested xenograft tumors with isotope labeled glucose injection. Micro-vessel density and percentage of apoptotic cells in the tumors were evaluated by immunohistochemistry using anti-CD34 antibody and anti-cleaved caspase 3 antibody, respectively. The effect of BIBF1120 on metabolic status in xenograft tumors was evaluated by imaging MS.
Result: Imaging MS revealed heterogeneity of metabolites distribution in the tumors before BIBF1120 treatment. In areas with rich ATP, many viable cells were seen with abundant glutathione, an anti-oxidant metabolite and glutamate, a metabolite derived from oxidative phosphorylation. Otherwise, other areas with rich lactate, a metabolite derived from glycolysis pathway, have less ATP and glutathione level compared with the areas with rich ATP. BIBF1120 inhibited growth of all of the xenograft tumors tested, although it did not directly show inhibitory effect for proliferation rate of those cells in vitro. BIBF1120-treated tumors exhibited significantly lower micro-vessel density compared to the vehicle-treated tumors (p&lt;0.05) and reduced 2-3 DPG (a maker of blood flow), glucose, and ATP levels. Lactate level was increased in BIBF1120-treated tumors. Imaging MS with isotope labeled glucose injection revealed that BIBF1120 treatment induced inactivation of oxidative phosphorylation pathway and activation of glycolysis pathway, especially in the peripheral areas of tumors.
Conclusion: We successfully visualized the heterogeneous distribution of metabolites in the tumors, and the effect of BIBF1120 on human lung cancer derived tumors. This is the first report to visualize the distribution of metabolites in engrafted tumors and the efficacy of BIBF1120 on the nutrition status of tumors.
Citation Format: Daisuke Arai, Hiroyuki Yasuda, Kenzo Soejima, Shizuko Kagawa, Junko Hamamoto, Shigenari Nukaga, Katsuhiko Naoki, Katsura Emoto, Yuki Sugiura, Makoto Suematsu, Tomoko Betsuyaku. Visualizing the distribution of metabolites and the efficacy of BIBF1120 on metabolic status of lung cancer derived tumors by imaging mass-spectrometry (MS). [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 4230.