MP61-04 ALTERATIONS IN CELL LINE SENSITIVITY TO SMALL MOLECULE INHIBITORS BETWEEN UROTHELIAL CANCER LINES AND THEIR METASTATIC DERIVATIVES (2016)

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology II1 Apr 2016MP61-04 ALTERATIONS IN CELL LINE SENSITIVITY TO SMALL MOLECULE INHIBITORS BETWEEN UROTHELIAL CANCER LINES AND THEIR METASTATIC DERIVATIVES Louis Krane, Reema Railkar, Iawan Hsu, Abhinav Sidana, Kai Hammerich, Cathy Vocke, Paul Meltzer, Craig Thomas, Marc Ferrer, and Piyush Agarwal Louis KraneLouis Krane More articles by this author , Reema RailkarReema Railkar More articles by this author , Iawan HsuIawan Hsu More articles by this author , Abhinav SidanaAbhinav Sidana More articles by this author , Kai HammerichKai Hammerich More articles by this author , Cathy VockeCathy Vocke More articles by this author , Paul MeltzerPaul Meltzer More articles by this author , Craig ThomasCraig Thomas More articles by this author , Marc FerrerMarc Ferrer More articles by this author , and Piyush AgarwalPiyush Agarwal More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.878AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Chemotherapy for urothelial cancer (UC) is less effective in patients with large volumes of metastatic disease, likely due to acquired driver mutations. These acquired driver mutation may lead to differences in sensitivity between primary cell lines and metastatic clones. We utilized quantitative high throughput screening (qHTS) to compare effectiveness of cell inhibition in two bladder cancer cell lines with their derived metastatic lines. METHODS We screened bladder cancer cell line T24 and derived cell lines (T24T, SLT3 and FL3) and UMUC-3 with derived cell lines (LUL2 ∖UMUC3-luc (LUC3)) against 1,912 oncology-focused drugs using a 48 hr cell proliferation assay with an ATP-based readout (CellTiterGlo), for activity and potency of the compounds in a dose response manner. We identified candidate drugs based on two parameters: 1) more than 70% inhibition at 48 hours and 2) a curve class of -1.1/-1.2 indicating curve class with good fit (r2>0.9). We then used a custom capture next generation DNA sequencing chip to compare exonic regions of 229 known cancer related genes between the primary and metastatic tumor cell lines. RESULTS The qHTS compounds are demonstrated in figure 1a for T24 and figure 1b for UMUC3, cell lines respectively. Using predefined criteria, only 141 of 1912 agents (7.4%) were fully active against T24. Yet, only 79 of these drugs (56%) continued to meet inclusion criteria for all T24 derived cell lines derived. Only 160 (8.4%) compounds met both criteria in the UMUC-3 line, of which 32 (20%) remained active in all derived cell lines. Exomic sequencing found RAC1 and CDH1 only mutated in UMUC3 derived lines and AR, CDK12, GATA1, GUCY1A2 for daughter lines of T24. CONCLUSIONS qHTS of a library of potential therapeutic interventions is a rapid and efficient way to produce a list of novel targets and therapies for bladder cancer. This data substantiate the differences in drug sensitivities between primary and metastatic tumors. Genetic differences between primary and metastatic lines including pathway alterations in the PI3K, RAS, MAPK, AR, glutamine transport and chromatin binding, may explain differences between sensitivities amongst derived lines. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e804 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Louis Krane More articles by this author Reema Railkar More articles by this author Iawan Hsu More articles by this author Abhinav Sidana More articles by this author Kai Hammerich More articles by this author Cathy Vocke More articles by this author Paul Meltzer More articles by this author Craig Thomas More articles by this author Marc Ferrer More articles by this author Piyush Agarwal More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...