Abstract
BackgroundCirculating 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. MethodsWe 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. ResultsThe 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. ConclusionsDLA 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 numberNL6019.
Highlights
Translational studies on advanced prostate cancer (PCa) have often been limited to static sources, such as resection of the primary tumour or biopsies [1e4]
We examined the impact of Diagnostic leukapheresis (DLA) density settings on Circulating tumour cell (CTC) enrichment in four patients, by performing the DLA procedure in two fractions of both 5L blood at 2% and 5% haematocrit, respectively
This study confirms that DLA is a safe and efficient method to harvest large amounts of CTCs from patients with metastatic PCa (mPCa)
Summary
Translational studies on advanced prostate cancer (PCa) have often been limited to static sources, such as resection of the primary tumour or biopsies [1e4]. CTCs can be obtained from peripheral blood in a relatively non-invasive manner, providing an accessible source of metastatic cells as an alternative to tumour biopsies. This is beneficial for patients with primarily bone metastasis such as in metastatic PCa (mPCa). The development of cancer organoids has allowed us to better capture the tumour-specific characteristics than standard 2D culture methods [11] Together, this provides the opportunity to use CTC derived organoids as a representative model of the current disease status and use for drug discovery and sensitivity-screening [12]. Our study provides an important step forward in implementing CTCs in individualised disease modelling, identifies several challenges that require further optimisation
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
