Abstract 3484: Ex-vivo cultures of freshly explanted tumor specimens (TIPCAN®), a potent translational approach for screening novel targeted agents

Abstract

Abstract Background: In vitro cell lines and in vivo mouse models are important screening tools for novel anticancer agents. However those preclinical models do not reliably reflect the complexity of human tumors and are, by essence, incapable of evaluating the intricate interplay of cancer cells in their native stroma. To evaluate novel drugs in hepatocellular (HCC) and Head and Neck squamous cell (HNSCC) carcinomas, we developed TIPCAN® as a novel approach using freshly explanted human tumor slices to test the pharmacodynamic effects of targeted agents or classic chemotherapies. Materials and Methods: Tumor samples were obtained from fresh surgical specimens and biopsies and cut in 300 µm thick slices using a Leica microtome. TIPCAN® was applied on tumor slices that were maintained in culture for 24-72 hours in a defined environment that allows for diffusion of oxygen and nutrient, in a specific culture medium. Viability of tissues was evaluated using different methods. Individual HCC and HNSCC tumor slices were exposed to various concentrations of targeted agents or chemotherapies. Tumor samples were then analyzed by immunohistochemistry and immunofluorescence for various pharmacodynamics. Slides were digitalized using an APERIO Scanscope system and biomarker expression was quantified using Image J Software. Results: We first established the suitable conditions for ex vivo culture, by testing several O2 levels and medium compositions. We then quality controlled TIPCAN® using over 30 liver tumors and defined HE staining as the most reliable tool to assess tissue quality and viability. Using appropriate biomarkers, we also confirmed that 24 to 72 hours of culture had minimal consequences on the potential phenotypical drift of HCC tumor slices, confirming the steadiness of our model, and allowing us to perform pharmacodynamics evaluation of several therapies. As an example, using our ex vivo approach we evaluated the consequence of sorafenib exposure on HCC tumors explants. We showed that sorafenib decreased HCC proliferation by almost 70%, induced a 2 to 8-fold increase in apoptosis and inhibited by 80% ERK and AKT phosphorylation, mimicking the reported effects of sorafenib in vivo. We then translated our technology in HNSCC, in which we tested the combination of carboplatin and everolimus and compared our results with clinical trial patients treated with the same drugs (CAPRA trial). Carboplatin and everolimus combination showed a decrease of S6 phosphorylation and MIB1 staining, and induction of caspase3, mimicking the effects observed in patients. Conclusion: Taken together, our results show that TIPCAN® can be used as a valuable tool to assess the pharmacodynamic effects of anticancer agents in HCC and HNSCC. TIPCAN® could contribute to evaluate drug effects in <72h in fresh human tumor tissues and could therefore be helpful for clinicians to select patients with HCC and HNSCC for intelligence medicine. Citation Format: Annemilai Tijeras-Raballand, Maria Serova, Miguel Albuquerque, Nathalie Colnot, pierre Bourgoin, Nelly Müller, Safi Dokmak, Mohamed Bouattour, Jacques Belghiti, Eric Raymond, Sandrine Faivre, Valérie Paradis, Armand de Gramont. Ex-vivo cultures of freshly explanted tumor specimens (TIPCAN®), a potent translational approach for screening novel targeted agents. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3484. doi:10.1158/1538-7445.AM2014-3484