Amin El-Heliebi1, Claudia Hille2, Navya Laxman3, Jessica Svedlund3, Christoph Haudum4,5, Erkan Ercan4, Thomas Kroneis4, Shukun Chen4, Maria Smolle5, Christopher Rossmann6, Tomasz Krzywkowski3, Annika Ahlford3,7, Evangelia Darai3, Gunhild von Amsberg8, Winfried Alsdorf8, Frank König9, Matthias Löhr10, Inge de Kruijff11, Sabine Riethdorf2, Tobias M Gorges2, Klaus Pantel2, Thomas Bauernhofer5,6, Mats Nilsson3, Peter Sedlmayr4. 1. Institute of Cell Biology, Histology and Embryology, Medical University Graz, Austria; amin.elheliebi@medunigraz.at. 2. Department of Tumor Biology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. 3. Science for Life Laboratory, Department of Biophysics and Biochemistry, Stockholm University, Solna, Sweden. 4. Institute of Cell Biology, Histology and Embryology, Medical University Graz, Austria. 5. Center for Biomarker Research in Medicine (CBmed); Graz, Austria. 6. Division of Oncology, Department of Internal Medicine, Medical University of Graz, Austria. 7. Devyser AB, Stockholm, Sweden. 8. Department of Hematology and Oncology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. 9. ATURO, Urology Practice, Berlin, Germany. 10. Center for Digestive Diseases, Karolinska University Hospital and Division of Surgery, CLINTEC, Karolinska Institutet, Stockholm, Sweden. 11. Erasmus MC Cancer Institute, Department of Medical Oncology and Cancer Genomics Netherlands, Rotterdam, the Netherlands.
Abstract
BACKGROUND: Liquid biopsies can be used in castration-resistant prostate cancer (CRPC) to detect androgen receptor splice variant 7 (AR-V7), a splicing product of the androgen receptor. Patients with AR-V7-positive circulating tumor cells (CTCs) have greater benefit of taxane chemotherapy compared with novel hormonal therapies, indicating a treatment-selection biomarker. Likewise, in those with pancreatic cancer (PaCa), KRAS mutations act as prognostic biomarkers. Thus, there is an urgent need for technology investigating the expression and mutation status of CTCs. Here, we report an approach that adds AR-V7 or KRAS status to CTC enumeration, compatible with multiple CTC-isolation platforms. METHODS: We studied 3 independent CTC-isolation devices (CellCollector, Parsortix, CellSearch) for the evaluation of AR-V7 or KRAS status of CTCs with in situ padlock probe technology. Padlock probes allow highly specific detection and visualization of transcripts on a cellular level. We applied padlock probes for detecting AR-V7, androgen receptor full length (AR-FL), and prostate-specific antigen (PSA) in CRPC and KRAS wild-type (wt) and mutant (mut) transcripts in PaCa in CTCs from 46 patients. RESULTS: In situ analysis showed that 71% (22 of 31) of CRPC patients had detectable AR-V7 expression ranging from low to high expression [1-76 rolling circle products (RCPs)/CTC]. In PaCa patients, 40% (6 of 15) had KRAS mut expressing CTCs with 1 to 8 RCPs/CTC. In situ padlock probe analysis revealed CTCs with no detectable cytokeratin expression but positivity for AR-V7 or KRAS mut transcripts. CONCLUSIONS: Padlock probe technology enables quantification of AR-V7, AR-FL, PSA, and KRAS mut/wt transcripts in CTCs. The technology is easily applicable in routine laboratories and compatible with multiple CTC-isolation devices.
BACKGROUND: Liquid biopsies can be used in castration-resistant prostate cancer (CRPC) to detect androgen receptor splice variant 7 (AR-V7), a splicing product of the androgen receptor. Patients with AR-V7-positive circulating tumor cells (CTCs) have greater benefit of taxane chemotherapy compared with novel hormonal therapies, indicating a treatment-selection biomarker. Likewise, in those with pancreatic cancer (PaCa), KRAS mutations act as prognostic biomarkers. Thus, there is an urgent need for technology investigating the expression and mutation status of CTCs. Here, we report an approach that adds AR-V7 or KRAS status to CTC enumeration, compatible with multiple CTC-isolation platforms. METHODS: We studied 3 independent CTC-isolation devices (CellCollector, Parsortix, CellSearch) for the evaluation of AR-V7 or KRAS status of CTCs with in situ padlock probe technology. Padlock probes allow highly specific detection and visualization of transcripts on a cellular level. We applied padlock probes for detecting AR-V7, androgen receptor full length (AR-FL), and prostate-specific antigen (PSA) in CRPC and KRAS wild-type (wt) and mutant (mut) transcripts in PaCa in CTCs from 46 patients. RESULTS: In situ analysis showed that 71% (22 of 31) of CRPC patients had detectable AR-V7 expression ranging from low to high expression [1-76 rolling circle products (RCPs)/CTC]. In PaCa patients, 40% (6 of 15) had KRAS mut expressing CTCs with 1 to 8 RCPs/CTC. In situ padlock probe analysis revealed CTCs with no detectable cytokeratin expression but positivity for AR-V7 or KRAS mut transcripts. CONCLUSIONS: Padlock probe technology enables quantification of AR-V7, AR-FL, PSA, and KRAS mut/wt transcripts in CTCs. The technology is easily applicable in routine laboratories and compatible with multiple CTC-isolation devices.
Authors: Irene Casanova-Salas; Alejandro Athie; Paul C Boutros; Marzia Del Re; David T Miyamoto; Kenneth J Pienta; Edwin M Posadas; Adam G Sowalsky; Arnulf Stenzl; Alexander W Wyatt; Joaquin Mateo Journal: Eur Urol Date: 2021-01-07 Impact factor: 24.267
Authors: Tae Hyun Kim; Yang Wang; C Ryan Oliver; Douglas H Thamm; Laura Cooling; Costanza Paoletti; Kaylee J Smith; Sunitha Nagrath; Daniel F Hayes Journal: Nat Commun Date: 2019-04-01 Impact factor: 14.919
Authors: Claudia Hille; Tobias M Gorges; Sabine Riethdorf; Martine Mazel; Thomas Steuber; Gunhild von Amsberg; Frank König; Sven Peine; Catherine Alix-Panabières; Klaus Pantel Journal: Cells Date: 2019-09-11 Impact factor: 6.600