Abstract 3919: Liquid biopsy derived organoids as a potential platform for personalized cancer therapy in metastatic prostate cancer
Authors: Mout Lisanne , van Dessel Lisanne F. , Kraan Jaco , de Jong Anouk C. , Neves Rui P. , Erkens-Schulze Sigrun , Siewerts Anieta M. , van Riet Job , de Wit Ronald , Sleijfer Stefan , Hamberg Paul , Sandberg Yorick , te Boekhorst Peter A. , van de Werken Harmen J. , Martens John W. , Stoecklein Nikolas H. , van Weerden Wytske M. , Lolkema Martijn P. 
Published: September 2020

Abstract
Introduction
Circulating tumor cells (CTCs) can serve as a source of metastatic tumor material, however their low numbers often limit downstream applications. Diagnostic leukapheresis (DLA) has been shown to substantially increase CTC yield. In this study we isolated CTCs from metastatic prostate cancer (mPCa) patients by DLA to propagate them in vitro as organoid cultures. Furthermore, tumor-derived organoids were used as a model for drug discovery and sensitivity-screening, thereby exploring potential treatment selection.
Methods
We included 44 mPCa patients into the study and 18 were selected for DLA, based on the presence of ≥5 CTCs/ 7.5 mL blood. We optimized the DLA procedure by comparing low versus high density settings and their impact on CTC isolation efficacy. As the DLA product contains a median of 8.6*10^9 white blood cells (WBC), stringent enrichment methods are needed. CTC enrichment from DLA product was performed by antibody-based WBC depletion alone, or combined with subsequent EpCAM based enrichment. Enriched CTC fractions were cultured in vitro under optimized conditions, to initiate organoid expansion.
Results
We show that DLA is a safe and efficient method to collect large amounts of CTCs from mPCa patients. With optimized DLA settings we were able to improve CTC enrichment and observed a non-significant increase in CTC yield from DLA (median CTC recovery 15339 vs 5796, P=0.125). WBC depletion alone was found to reduce WBCs by ~2000-fold while retaining &gt;50% of the CTCs, resulting in a WBC to CTC ratio of 545:1. We were able to culture and confirm CTC-derived organoids in 9/18 samples, including one organoid cell line, EMC-PCa-41. Whole Genome Sequencing (WGS) of EMC-PCa-41 revealed a triploid genome characterized by focal amplification of AR, a TMPRSS2-ERG fusion, a PTEN deletion and multiple inter-chromosomal rearrangements. Next we determined copy number profiles in single CTCs and matched organoids from two patients using shallow WGS. These data confirm prior data that CTCs represent the inherent intra-patient heterogeneity and organoids resemble CTCs from the original DLA product. Moreover, we performed an in vitro drug screen with the organoid cell line EMC-PCa-41, and found that it has a limited response to Enzalutamide, which correlated with the relatively short response to Enzalutamide that was observed in the patient.
Conclusion
Overall our study demonstrates that DLA provides a high CTC yield which enables short-term organoid cultures that preserve the genomic hallmarks of prostate cancer. Viable CTCs obtained by DLA may serve as a (personalized) drug screening system in metastatic prostate cancer.
Citation Format: Lisanne Mout, Lisanne F. van Dessel, Jaco Kraan, Anouk C. de Jong, Rui P. Neves, Sigrun Erkens-Schulze, Anieta M. Siewerts, Job van Riet, Ronald de Wit, Stefan Sleijfer, Paul Hamberg, Yorick Sandberg, Peter A. te Boekhorst, Harmen J. van de Werken, John W. Martens, Nikolas H. Stoecklein, Wytske M. van Weerden, Martijn P. Lolkema. Liquid biopsy derived organoids as a potential platform for personalized cancer therapy in metastatic prostate cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3919.