Abstract LB-282: Ex vivo 3d drug response profiling of primary human ovarian cancer differentiates treatment-naive and relapsed patients and molecular subtypes
Authors: Desrochers Tessa , Shuford Stephen , Mattingly Christina , Holmes Lillia , Gevaert Matt , Elder Jeff , Orr David , Corless Christopher , Puls Larry , Crosswell Hal E. 
Published: August 2015

Background: Epithelial ovarian cancer (EOC) affects nearly 22,000 women annually and is the leading cause of death from gynecologic cancer in the United States. Five year cure rates are &lt;40% and approximately 14,000 will die each year. Large-scale efforts are currently underway to use molecular profiling via next generation sequencing (NGS) technology to guide treatment in cancer patients with poor prognosis, but limited application of NGS in ovarian cancer has been reported. In this report, our previously described Ex Vivo 3D Drug Response Profiling was used to identify response differences between newly diagnosed and relapsed ovarian cancer patients and correlated with NGS of primary tissue. Materials & Methods: Processing of Ovarian Cancers: Under informed consent, ovarian cancer samples were obtained and processed using standard mincing & digestion. 3D spheroids were developed and 3D perfused Ovarian Microtumors were cultured using the 3DKUBE™. Ex Vivo Testing & Analysis: Cultured cells were exposed to clinically relevant concentrations of cytotoxic or targeted agents. Relative IC50s and total percent inhibition were used for ranking compounds. Isolated DNA was sequenced in a CLIA laboratory using a 37-gene NGS panel (GeneTrails®) on the Ion Torrent PGM. Results: Tissues from both newly diagnosed, treatment naive subjects and relapsed subjects were obtained and processed after IRB-approved tissue consent was provided. Spheroid formation was uniform across all malignant tumor types. There was a statistical difference for 3D spheroids treated with Carboplatin formed from either naïve or relapse tissue. The relapse samples had a significantly higher median IC50 than did the naïve samples (70.8 vs. 17.4). This significant difference was not apparent in matched 2D treatment groups. Gemcitabine response varied across tissue type, but did not correlate with traditional biomarkers (i.e. hENT mRNA expression). NGS testing turn-around time was a median of 9 days (range 7-14). Ovarian 3D microtumors were successfully perfuse and tested with targeted agents guided by NGS results, as evidence by a tumor with a mutation of EGFR (p.P265T, clinical significance unknown) with both erlotinib and afatanib demonstrating activity (3.3uM and 0.7uM, respectively).

Conclusions: EV3D DRP successfully differentiates carboplatin response and 3D perfusion of microtumors permits ex vivo culture of primary ovarian samples for genotypic-phenotypic response determination. Clinical response data and clinical correlation is ongoing. EV3D DRP permits phenotypic drug response correlation with molecular profiling in real time and may be a clinically relevant functional assay for driver mutation identification and maximal patient response to targeted agent(s).

Citation Format: Tessa Desrochers, Stephen Shuford, Christina Mattingly, Lillia Holmes, Matt Gevaert, Jeff Elder, David Orr, Christopher Corless, Larry Puls, Hal E. Crosswell. Ex vivo 3d drug response profiling of primary human ovarian cancer differentiates treatment-naive and relapsed patients and molecular subtypes. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-282. doi:10.1158/1538-7445.AM2015-LB-282