AIRC Start-Up Unit Pisa, Lab-unit: Dr. Niccola Funel, Dr. Elisa Giovannetti (PI)Surgery-unit: Dr. N De Lio, Dr. V Perrone, Prof. U Boggi;

Oncology-unit: Dr. S Caponi, Dr. E Vasile; Pathology-Unit: Dr. LE Pollina, Prof. D Campani; VUmc Amsterdam collaborators: Dr. T Würdinger, Prof. GJ Peters

www.cancerpharmacology.org

Bioluminescent pancreatic cancer mouse models from genetically characterized primary cells: a platform for drug discovery and toward personalized treatment in pancreatic cancer

PRELIMINARY RESULTS

- To develop orthotopic mouse models (Fig.1) from primary pancreatic tumor cells and optimize bioluminescent imaging- To test new targeted drugs in vitro and in vivo

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Fig. 2. A-B) Fluc (red curves) and Gluc (blue curves) activities of the PDAC-1/2-FM-GC mouse models. The insets show representative fluorescence microscopy images of PDAC-1-FM-GC cells; C) Kaplan-meier survival curves; (D) Magnetic resonance imaging (MRI)-images and (E) high-frequency ultrasound in 3D Power Doppler

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Fig.3. Infiltration (A, F, arrows,) PDAC-associated desmoplastic reaction (B, G,), well-defined glandular pattern, with differentiated duct formations (C, H, arrows,), and inflammatory reaction (D, I, arrows,). IHC for (K) Ck 8/18, (L) CK7, (M) EGFR, (N) CEA; (O,Q); liver, and (P,R) lung metastases Fig. 4. H:human; C:cells; M:mouse

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Fig. 6. A) Growth inhibitory effect of c-Met inhibitors, crizotinib, tivantinib and DN-30 in PDAC-3 cells; inset, modulation of protein expression of c-Met and phospho-c-Met; B) Growth inhibitory effect of the c-Met inhibitor crizotinib in PDAC-1/2/3/4 cells, as determined by SRB assay; C) Growth inhibitory effect of crizotinib, gemcitabine and their combination in PDAC-3 cells; D) Intracellular accumulation of gemcitabine after exposure to 10 μM gemcitabine for 4 hours, as determined by LC-MS/MS

Fig. 7. A) Fluc activity in orthotopic PDAC-3-FM-GC mice models (n=5 mice per group), treated with 25 mg/kg crizotinib (in DMSO and 0.5% methylcellulose, orally), 100 mg/kg gemcitabine (i.p.), q3d×4;B) Survival analysis in the PDAC-3-FM-GC mice; C-D) Gemcitabine and crizotinib concentration in circulating blood and tissue samples of the PDAC-3-FM-GC mice treated with gemcitabine and crizotinib, as determined by LC-MS/MS

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PDAC-FM-GC mouse models

Histopathology and immunohistopathology of the orthotopic PDAC mouse models

Genetic characteristics of the orthotopic PDAC mouse model compared to their human originator tumors (array-CGH)

C-Met overexpression

Fig. 5. A) Aberrations in the chromosome 7 of the originator human specimen, carrying 7q31.1 cytoband copy gain; B) MET copy number determined by quantitative PCR; C) MET mRNA expression; D) Representative pictures of patient tumors, and primary cells stained with c-Met

Synergism of crizotinib and gemcitabine: in vitro

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Synergism of crizotinib and gemcitabine: in vivo

Our orthotopic PDAC imaging models showed genetic, histopathological and metastatic features similar to their originator tumors One of our models identified c-Met as a potential therapeutic target

CONCLUSIONS & FUTURE STUDIES Crizotinib and gemcitabine were synergistic in vitro and in vivoCrizotinib increased blood, tumor cell and tissue concentrations of gemcitabineThese models provide a platform to test the efficacy of targeted innovative anticancer drugs