Abstract 5545: Optimal drug concentration screening and evaluation in cancer stem cells & 3D tumor stem cell cultures drug response assays in association with clinical efficacy for pancreatic cancer stem cell.
Authors: Clery John P. , Gomez Esteban , Sharma Michael , khemani Aabha , Sharma Cristian , Punzalan Rubio , navel miriam , Amezcua Natalee , Jani Jitesh , Sharma Jay 
Published: August 2014

Three-dimensional (3D) culture of cancer cell lines has long been advocated as a better model of the malignant phenotype that is most closely related to tumorigenicity in vivo. Moreover, new drug development requires simple in vitro models that resemble the in vivo situation more in order to select active drugs against solid tumors and to decrease the use of experimental animals. The induction of chemotherapy or concomitant chemoradiotherapy has been used to increasingly to improve survival, and organ preservation. This approach encounters significant morbidity and mortality. Therefore reliable chemosensitivity assays are needed to accurately predict the response to chemotherapy and guide the selection and treatment of cancer patients. The purpose of this study is to examine and evaluate optimum drug candidates in vitro chemosensitivity on patient tumor tissues directly in culture and on their Cancer stem cell cultures. The tumor samples obtained after surgery or biopsy, were placed immediately in Celprogen Tumor Transportation Media and shipped at 4-8 OC for processing. Tissues were washed with 1X PBS solution and aseptically cut into 0.5mm sections and cultured in 6 well tissue culture plates with an insert pre-coated with ECM. All cancer cell types remain viable and maintain their native architecture for at least 14 days and incorporated DNA measured by adding EdU(5-ethynyl-2’-deoxyuridine) to the culture. The efficacy of various therapeutic agents targeting major pathways (wnt,Notch,PI3K,MAPK,STAT) and chemotherapy agents were tested using DNA uptake and TUNNEL assay anti-cancer agents was calculated according to the inhibition index. The same compounds were tested for utilizing the patient's Pancreatic Cancer Stem Cell Cultures established with Celprogen's Media and ECM. Expression of PDX-1, SHH, CD24, CD44, CD133, EpCAM, CBX7, OCT4, SNAIL, SLUG, TWIST, Ki-67, E-cadherin, β-catenin and vimentin were quantified by qPCR or immunocytochemistry per cell culture. The epithelial-mesenchymal transition (EMT) is linked to induction of a stem-cell like phenotype. We cultured the cells in low oxygen since Tumor hypoxia induces EMT, which induces invasion and metastasis, and is linked to cancer stem cells (CSCs). Among the 600 compounds tested Gemcitabine, Taxol, Fluorouracil, Leucovorin, Irinotecan, and Oxaliptin were not effective against Pancreatic Cancer Stem cell (CSC) but were effective on tumor cells (differentiated CSCs). We were able to show 6 compounds that were effective against Pancreatic CSC targeting selected pathways.

Citation Format: John P. Clery, Esteban Gomez, Michael Sharma, Aabha khemani, Cristian Sharma, Rubio Punzalan, miriam navel, Natalee Amezcua, Jitesh Jani, Jay Sharma. Optimal drug concentration screening and evaluation in cancer stem cells & 3D tumor stem cell cultures drug response assays in association with clinical efficacy for pancreatic cancer stem cell. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5545. doi:10.1158/1538-7445.AM2013-5545