Abstract 644: Utilizing a novel luciferase labeling technique to establish and validate preclinical models of pancreatic cancer
Authors: Bernoulli Jenni , Bozza Matthias , Fagerlund Katja M. , Tuomela Johanna , Suominen Mari I. , Dilly Suzanne , Morris George , Halleen Jussi M. , Harbottle Richard 
Published: July 2016

Bioluminescent-labeling imaging (BLI) allows sensitive non-invasive sequential imaging of tumor development and early metastasis. Current methods for the genetic modification of cells typically use integrating genotoxic viruses that can disrupt the molecular behavior of cancer cell lines due to their random nature of integration. A primary aim of the study was to utilize a non-integrating DNA vector that comprises an S/MAR (Scaffold/Matrix Attachment Region) element to stably genetically modify pancreatic cancer cells to persistently express the reporter gene luciferase without altering the molecular behavior of the cell or altering its sensitivity to therapeutic drug treatments. Once a novel isogenic cell line is generated the cells can subsequently be used in xenograft studies. A second aim was to validate these established models with gemcitabine and test the efficacy of VAL401, formulation of risperidone in rumenic acid. Human BxPC3, Capan-1, MiaPaCa-2 and Panc-1 pancreatic cancer cells were stably transfected with a pSMARt-UBC-Luc DNA vector and cultured for 4 weeks under selection. Colonies that formed after this period were isolated and expanded in normal medium and evaluated for luciferase expression and the molecular integrity of the DNA vector. Efficacy of gemcitabine was tested in these new luciferase expressing cell lines and VAL401 was tested in Capan-1-luc cells. For in vivo studies, BxPC3-luc cells were inoculated orthotopically into the pancreas of athymic nude mice and stratified into groups: control, gemcitabine, VAL401 (1mg/kg, p.o. daily) and VAL401 (2mg/kg, p.o. daily). In vitro validation results indicated that the luciferase transfected cells maintained their original properties with stable expression. Gemcitabine inhibited cell proliferation in all established cell lines. VAL401 inhibited cell proliferation of Capan-1-luc cells at 50 μM concentration. BxPC3-luc cells inoculated orthotopically into the pancreas were followed for 5 weeks with BLI by IVIS, and the results demonstrated high-quality follow-up of tumor growth. BxPC3-luc cells induced growth of pancreatic tumors with high take rate in all groups. Gemcitabine and both studied doses of VAL401 decreased tumor volume, and the same trend was seen in tumor weight and the BLI during the study. In conclusion, both gemcitabine and VAL401 decreased tumor volume and the same trend was observed using BLI. Our results demonstrated that S/MAR DNA vectors are able to produce genetically modified cells without the limitations of random genomic integration, whilst providing extra-chromosomal mitotic stability and high levels of sustained transgene expression. When utilized in orthotopic xenograft studies, these luciferase expressing cells formed a reliable and essentially non-invasive imaging platform that substantially improves the efficacy of testing anticancer drug candidates.

Citation Format: Jenni Bernoulli, Matthias Bozza, Katja M. Fagerlund, Johanna Tuomela, Mari I. Suominen, Suzanne Dilly, George Morris, Jussi M. Halleen, Richard Harbottle. Utilizing a novel luciferase labeling technique to establish and validate preclinical models of pancreatic cancer. [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 644.