Lisanne Mout1, Lisanne F van Dessel2, Jaco Kraan2, Anouk C de Jong3, Rui P L Neves4, Sigrun Erkens-Schulze5, Corine M Beaufort2, Anieta M Sieuwerts3, Job van Riet6, Thomas L C Woo3, Ronald de Wit3, Stefan Sleijfer3, Paul Hamberg7, Yorick Sandberg8, Peter A W Te Boekhorst9, Harmen J G van de Werken10, John W M Martens2, Nikolas H Stoecklein4, Wytske M van Weerden5, Martijn P Lolkema11. 1. Department of Medical Oncology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands; Department of Urology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands. 2. Department of Medical Oncology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands; Cancer Genomics Netherlands, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands. 3. Department of Medical Oncology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands. 4. Department of General, Visceral and Pediatric Surgery, Heinrich-Heine University, University Hospital Düsseldorf, Düsseldorf, Germany. 5. Department of Urology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands. 6. Department of Medical Oncology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands; Department of Urology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands; Cancer Computational Biology Center, Erasmus MC Cancer Institute, University Medical Center, Rotterdam, the Netherlands. 7. Department of Internal Medicine, Franciscus Gasthuis & Vlietland, Rotterdam, the Netherlands. 8. Department of Internal Medicine, Maasstad Hospital, Rotterdam, the Netherlands. 9. Department of Hematology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands. 10. Department of Urology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands; Cancer Computational Biology Center, Erasmus MC Cancer Institute, University Medical Center, Rotterdam, the Netherlands. 11. Department of Medical Oncology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, the Netherlands. Electronic address: m.lolkema@erasmusmc.nl.
Abstract
BACKGROUND: Circulating tumour cell (CTC)-derived organoids have the potential to provide a powerful tool for personalised cancer therapy but are restrained by low CTC numbers provided by blood samples. Here, we used diagnostic leukapheresis (DLA) to enrich CTCs from patients with metastatic prostate cancer (mPCa) and explored whether organoids provide a platform for ex vivo treatment modelling. METHODS: We prospectively screened 102 patients with mPCa and performed DLA in 40 patients with ≥5 CTCs/7.5 mL blood. We enriched CTCs from DLA using white blood cell (WBC) depletion alone or combined with EpCAM selection. The enriched CTC samples were cultured in 3D to obtain organoids and used for downstream analyses. RESULTS: The DLA procedure resulted in a median yield of 5312 CTCs as compared with 22 CTCs in 7.5 mL of blood. Using WBC depletion, we recovered 46% of the CTCs, which reduced to 12% with subsequent EpCAM selection. From the isolated and enriched CTC samples, organoid expansion succeeded in 35%. Successful organoid cultures contained significantly higher CTC numbers at initiation. Moreover, we performed treatment modelling in one organoid cell line and identified substantial tumour heterogeneity in CTCs using single cell DNA sequencing. CONCLUSIONS: DLA is an efficient method to enrich CTCs, although the modest success rate of culturing CTCs precludes large scale clinical application. Our data do suggest that DLA and subsequent processing provides a rich source of viable tumour cells. Therefore, DLA offers a promising alternative to biopsy procedures to obtain sufficient number of tumour cells to study sequential samples in patients with mPCa. TRIAL REGISTRATION NUMBER: NL6019.
BACKGROUND: Circulating tumour cell (CTC)-derived organoids have the potential to provide a powerful tool for personalised cancer therapy but are restrained by low CTC numbers provided by blood samples. Here, we used diagnostic leukapheresis (DLA) to enrich CTCs from patients with metastatic prostate cancer (mPCa) and explored whether organoids provide a platform for ex vivo treatment modelling. METHODS: We prospectively screened 102 patients with mPCa and performed DLA in 40 patients with ≥5 CTCs/7.5 mL blood. We enriched CTCs from DLA using white blood cell (WBC) depletion alone or combined with EpCAM selection. The enriched CTC samples were cultured in 3D to obtain organoids and used for downstream analyses. RESULTS: The DLA procedure resulted in a median yield of 5312 CTCs as compared with 22 CTCs in 7.5 mL of blood. Using WBC depletion, we recovered 46% of the CTCs, which reduced to 12% with subsequent EpCAM selection. From the isolated and enriched CTC samples, organoid expansion succeeded in 35%. Successful organoid cultures contained significantly higher CTC numbers at initiation. Moreover, we performed treatment modelling in one organoid cell line and identified substantial tumour heterogeneity in CTCs using single cell DNA sequencing. CONCLUSIONS: DLA is an efficient method to enrich CTCs, although the modest success rate of culturing CTCs precludes large scale clinical application. Our data do suggest that DLA and subsequent processing provides a rich source of viable tumour cells. Therefore, DLA offers a promising alternative to biopsy procedures to obtain sufficient number of tumour cells to study sequential samples in patients with mPCa. TRIAL REGISTRATION NUMBER: NL6019.
Authors: Maria Laura De Angelis; Federica Francescangeli; Chiara Nicolazzo; Michele Signore; Alessandro Giuliani; Lidia Colace; Alessandra Boe; Valentina Magri; Marta Baiocchi; Antonio Ciardi; Francesco Scarola; Massimo Spada; Filippo La Torre; Paola Gazzaniga; Mauro Biffoni; Ruggero De Maria; Ann Zeuner Journal: J Exp Clin Cancer Res Date: 2022-03-08
Authors: Manuel Ramirez-Garrastacho; Cristina Bajo-Santos; Jesus Martinez de la Fuente; Maria Moros; Carolina Soekmadji; Kristin Austlid Tasken; Aija Line; Elena S Martens-Uzunova; Alicia Llorente Journal: Br J Cancer Date: 2021-11-22 Impact factor: 7.640
Authors: Annelies Van Hemelryk; Lisanne Mout; Sigrun Erkens-Schulze; Pim J French; Wytske M van Weerden; Martin E van Royen Journal: Biomolecules Date: 2021-10-22
Authors: I E de Kruijff; A M Sieuwerts; N Beije; W J C Prager-van der Smissen; L Angus; C M Beaufort; M N Van; E Oomen-de Hoop; A Jager; P Hamberg; F E de Jongh; J Kraan; J W M Martens; S Sleijfer Journal: Front Oncol Date: 2021-06-25 Impact factor: 6.244