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Li H, Li P, Li F, Wang T. Application of CellSearch technique in detection of peripheral blood circulating tumour cell count in patients with head and neck cancer and its association with prognosis. Oncol Lett 2025; 29:100. [PMID: 39717066 PMCID: PMC11664308 DOI: 10.3892/ol.2024.14846] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/23/2024] [Accepted: 09/12/2024] [Indexed: 12/25/2024] Open
Abstract
The aim of the present study was to employ CELLSEARCH® technology for the detection of circulating tumor cells (CTCs) in the peripheral blood of head and neck cancer (HNC) patients, and to assess the association between CTC count and patient prognosis. In this retrospective study, a cohort of 56 patients diagnosed with HNC and receiving treatment at the Department of Otolaryngology, Head and Neck Surgery (Beijing Tongren Hospital) between December 2013 and June 2018 were selected. Utilizing CELLSEARCH® technology, the presence of CTCs were detected in samples of peripheral blood from patients with head and neck cancer (HNC) patients, and CTC counts were documented. CTC positivity was defined as CTCs ≥1/7.5 ml of peripheral blood. Comprehensive data encompassing general demographic profiles, pathological classifications, tumor node metastasis (TNM) staging, tumor histology and treatment modalities were gathered for each participant. The study employed the Kaplan-Meier method to scrutinize and compare survival rates between CTC-positive and CTC-negative cohorts, while both univariate and multivariate Cox regression analyses were conducted to discern the factors impacting the overall survival (OS) of individuals diagnosed with HNC. Out of the 56 patients examined, 14 individuals exhibited detectable levels of CTCs, resulting in a positivity rate of 25%. The analysis revealed a significant association between the levels of CTCs in patients with HNC and the utilization of non-surgical treatment (P<0.05), while no substantial associations were observed concerning sex, age, smoking habits, alcohol consumption, pathological classifications, TNM staging, tumor attributes and surgical interventions (all P>0.05). Survival analysis revealed a reduction in the OS among patients with HNC harboring CTC positivity in contrast to their CTC-negative counterparts. The comprehensive multivariate Cox regression analysis underscored the independent prognostic impact of CTC presence (HR=1.274; 95% CI, 1.119-1.451; P<0.001) and the implementation of non-surgical treatment (HR=0.268; 95% CI, 0.119-0.607; P=0.002) on the prognosis of individuals grappling with HNC. In conclusion, the levels of CTCs were an independent factor affecting outcomes in patients with HNC, with CTC-positive patients showing significantly shorter survival compared with CTC-negative cases.
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Affiliation(s)
- Haiyang Li
- Department of Otolaryngology, Beijing Daxing District People's Hospital, Beijing 102600, P.R. China
| | - Pingdong Li
- Department of Otolaryngology, Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, P.R. China
- Key Laboratory of Otorhinolaryngology, Head and Neck Surgery, Ministry of Education, Beijing Institute of Otorhinolaryngology, Beijing 100730, P.R. China
| | - Feng Li
- Department of Otolaryngology, Beijing Daxing District People's Hospital, Beijing 102600, P.R. China
| | - Tao Wang
- Department of Otolaryngology, Beijing Daxing District People's Hospital, Beijing 102600, P.R. China
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Chang JL, Huang CJ, Tsai YC, Chiang NJ, Huang YS, Hung SC, Shan YS, Lee GB. An integrated microfluidic system for automatic detection of cholangiocarcinoma cells from bile. LAB ON A CHIP 2024; 24:375-382. [PMID: 38126571 DOI: 10.1039/d3lc00862b] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/23/2023]
Abstract
Cholangiocarcinoma (CCA) is an aggressive cancer that originates from the epithelial cells lining the bile ducts. Due to its location deep within the body and nonspecific symptoms in the early stages, it is often diagnosed at the advanced stage, thus leading to worse prognosis. Circulating tumor cells within liquid biopsies (i.e. blood) have been considered as promising biomarkers for CCA diagnosis, though current methods for profiling them are not satisfactory in terms of sensitivity and specificity. Herein we developed a new cancer cell probing and immuno-tracking assay known as "CAPTURE", which was performed on an integrated microfluidic system (IMS) to automate CCA diagnosis from bile with a sample amount of only 1 mL. The assay utilized magnetic beads surface-coated with two affinity reagents, a nucleic acid aptamer (HN16) and a glycosaminoglycan (SCH 45-mix), for capturing cancer cells in bile; the "gold standard" anti-epithelial cell adhesion molecule was used as a comparison. In a single-blind test of 54 CCA-positive (+) and 102 CCA-negative (-) clinical samples, sensitivities and specificities of 96 and 80%, respectively, were documented with the CAPTURE assay on-bench. An IMS composed of a centrifugal module for sample pretreatment and a CAPTURE module for cell capture and staining was integrated with a new "vertical integration module" for detecting cancer cells from bile without human intervention. Furthermore, a novel micro-tier structure within the centrifugal module was designed to block passage of gallbladder stones with diameters >1 mm, thereby preventing their interference during the subsequent CAPTURE assay. Improved sensitivity and specificity (100 & 83%, respectively) by using three affinity reagents were achieved on the IMS when using 26 clinical bile samples, confirming its clinical bio-applicability for CCA diagnosis. This approach could be therefore used for early-stage CCA diagnostics, ideally enabling effective treatment, as well as reducing potential for relapse.
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Affiliation(s)
- Jui-Lin Chang
- Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
| | - Chien-Jui Huang
- Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan
| | - Yi-Cheng Tsai
- Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan
| | - Nai-Jung Chiang
- Department of Oncology, Taipei Veterans General Hospital, Taipei, Taiwan.
| | - Yu-Shan Huang
- Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan
| | | | - Yan-Shen Shan
- Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan
| | - Gwo-Bin Lee
- Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
- Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan
- Institute of NanoEngineering and MicroSystems, National Tsing Hua University, Hsinchu, Taiwan
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Stevens M, Mentink A, Nanou A, Coumans FAW, Isebia KT, Kraan J, Hamberg P, Martens JWM, Terstappen LWMM. Improved enrichment of circulating tumor cells from diagnostic leukapheresis product. Cytometry A 2023; 103:881-888. [PMID: 37461156 DOI: 10.1002/cyto.a.24779] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/09/2023] [Revised: 06/22/2023] [Accepted: 07/11/2023] [Indexed: 08/03/2023]
Abstract
The median number of circulating tumor cells (CTCs) detected in 7.5 mL of peripheral blood by CellSearch (PB-CS) in patients with metastatic prostate cancer is in the order of 1-10, which means many samples have insufficient tumor cells for comprehensive characterization. A significant increase is obtained through diagnostic leukapheresis (DLA), however, only 2%-3% of the DLA product can be processed per CellSearch test, limiting the gain. We processed aliquots from 30 DLA products of metastatic prostate cancer patients consisting of 0.2 × 109 leukocytes using CellSearch (DLA-CS) as well as the newly introduced reduced enrichment reagent protocol (RER), which uses 10-fold less enrichment reagents than DLA-CS. The number of tumor cells and the total number of captured cells were determined using the CellTracks Analyzer. Additionally, for six DLA samples, a 1.0 × 109 leukocyte aliquot was processed (RER+), using twofold less enrichment reagents than DLA-CS. A median 2.7-fold reduction in leukocyte co-enrichment was found between DLA-CS and RER methods without any loss in tumor cell recovery (Wilcoxon Signed Ranks Test, p = 0.953). Using 1.0 × 109 leukocyte aliquots a fourfold increase in tumor cells was found compared to DLA-CS and a 19-fold increase compared to PB-CS was obtained. The here-introduced RER protocol results in a higher final sample purity without any loss in tumor cell recovery while using 10-fold less CellSearch capture reagent. With this improved method, 26% of the leukapheresis sample can now be processed using reagents from a single CellSearch test, enabling the obtainment of a sufficient number of CTCs for comprehensive characterization in most metastatic prostate cancer patients.
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Affiliation(s)
- Michiel Stevens
- Department of Medical Cell BioPhysics, Faculty of Science and Technology, University of Twente, Enschede, The Netherlands
| | - Anouk Mentink
- Department of Medical Cell BioPhysics, Faculty of Science and Technology, University of Twente, Enschede, The Netherlands
| | - Afroditi Nanou
- Department of Medical Cell BioPhysics, Faculty of Science and Technology, University of Twente, Enschede, The Netherlands
| | - Frank A W Coumans
- Department of Medical Cell BioPhysics, Faculty of Science and Technology, University of Twente, Enschede, The Netherlands
| | - Khrystany T Isebia
- Department of Medical Oncology, Erasmus MC Cancer Institute, Rotterdam, The Netherlands
| | - Jaco Kraan
- Department of Medical Oncology, Erasmus MC Cancer Institute, Rotterdam, The Netherlands
| | - Paul Hamberg
- Department of Oncology, Franciscus Gasthuis & Vlietland Hospital, Schiedam, The Netherlands
| | - John W M Martens
- Department of Medical Oncology, Erasmus MC Cancer Institute, Rotterdam, The Netherlands
| | - Leon W M M Terstappen
- Department of Medical Cell BioPhysics, Faculty of Science and Technology, University of Twente, Enschede, The Netherlands
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Mentink A, Isebia KT, Kraan J, Terstappen LWMM, Stevens M. Measuring antigen expression of cancer cell lines and circulating tumour cells. Sci Rep 2023; 13:6051. [PMID: 37055551 PMCID: PMC10101999 DOI: 10.1038/s41598-023-33179-y] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/02/2022] [Accepted: 04/08/2023] [Indexed: 04/15/2023] Open
Abstract
When evaluating EpCAM-based enrichment technologies for circulating tumour cells (CTCs), the cell lines used should closely resemble real CTCs, meaning the EpCAM expression of CTCs needs to be known, but also the EpCAM expression of cell lines at different institutions and times is important. As the number of CTCs in the blood is low, we enriched CTCs through the depletion of leukocytes from diagnostic leukapheresis products of 13 prostate cancer patients and measured EpCAM expression using quantitative flow cytometry. Antigen expression was compared between multiple institutions by measuring cultures from each institution. Capture efficiency was also measured for one of the used cell lines. Results show CTCs derived from castration-sensitive prostate cancer patients have varying but relatively low EpCAM expression, with median expression per patient ranging from 35 to 89,534 (mean 24,993) molecules per cell. A large variation in the antigen expression of identical cell lines cultured at different institutions was found, resulting in recoveries when using the CellSearch system ranging from 12 up to 83% for the same cell line. We conclude that large differences in capture efficiency can occur while using the same cell line. To closely resemble real CTCs from castration-sensitive prostate cancer patients, a cell line with a relatively low EpCAM expression should be used, and its expression should be monitored frequently.
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Affiliation(s)
- Anouk Mentink
- Medical Cell Biophysics Group, Techmed Center, Faculty of Science and Technology, University of Twente, PO Box 217, 7500 AE, Enschede, The Netherlands
| | - Khrystany T Isebia
- Department of Medical Oncology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, The Netherlands
| | - Jaco Kraan
- Department of Medical Oncology, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, The Netherlands
| | - Leon W M M Terstappen
- Medical Cell Biophysics Group, Techmed Center, Faculty of Science and Technology, University of Twente, PO Box 217, 7500 AE, Enschede, The Netherlands
| | - Michiel Stevens
- Medical Cell Biophysics Group, Techmed Center, Faculty of Science and Technology, University of Twente, PO Box 217, 7500 AE, Enschede, The Netherlands.
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Xing L, Wan X, Yu MT, He YJ, Wang Y, Zhou TJ, Liu XY, Sun Y, Luo J, Wang WJ, Jiang HL. A novel whole blood purifier for efficient capture and separation of circulating tumor cells. Biosens Bioelectron 2023; 232:115292. [PMID: 37062202 DOI: 10.1016/j.bios.2023.115292] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/20/2023] [Revised: 03/23/2023] [Accepted: 03/30/2023] [Indexed: 04/03/2023]
Abstract
Circulating tumor cells (CTCs) as important biomarkers for noninvasive clinical diagnosis and prognostic evaluation are significant in predicting the overall survival and progression-free survival of cancer patients. However, the current typical CTCs separation and enrichment techniques were limited to a single collection of small-volume blood samples, which was inadequate to comprehensively profile the distribution of CTCs in the systemic blood. In addition, those techniques cannot reduce metastasis of CTCs unless adjuvant chemotherapy. Herein, inspired by hemodialysis, we designed a whole blood purifier (WBP) composed of a functionalized special spiral-like glass tube modified by anti-epithelial cell adhesion molecule (anti-EpCAM). The WBP allowed real-time capture, enrichment and removal of CTCs from systemic blood circulation, and the purified blood was immediately returned to the body. Furthermore, the WBP did not cause any organic damages in vivo. This approach achieves the high accuracy of liquid biopsy technology and is expected to become an effective clinical adjuvant therapy for tumor metastasis.
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Affiliation(s)
- Lei Xing
- State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, 210009, China; Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, China Pharmaceutical University, Nanjing, 210009, China; Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases, China Pharmaceutical University, Nanjing, 210009, China
| | - Xing Wan
- State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, 210009, China
| | - Ming-Tao Yu
- State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, 210009, China
| | - Yu-Jing He
- Institute of Pharmacology, School of Pharmaceutical Sciences, Shandong First Medical University & Shandong Academy of Medical Sciences, Taian, 271000, Shandong, China
| | - Yi Wang
- State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, 210009, China
| | - Tian-Jiao Zhou
- State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, 210009, China
| | - Xiao-Ying Liu
- State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, 210009, China
| | - Ying Sun
- State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, 210009, China
| | - Jun Luo
- State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, 210009, China
| | - Wen-Jia Wang
- State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, 210009, China
| | - Hu-Lin Jiang
- State Key Laboratory of Natural Medicines, Department of Pharmaceutics, China Pharmaceutical University, Nanjing, 210009, China; Jiangsu Key Laboratory of Druggability of Biopharmaceuticals, China Pharmaceutical University, Nanjing, 210009, China; Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases, China Pharmaceutical University, Nanjing, 210009, China.
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Verkhovskii R, Ermakov A, Grishin O, Makarkin MA, Kozhevnikov I, Makhortov M, Kozlova A, Salem S, Tuchin V, Bratashov D. The Influence of Magnetic Composite Capsule Structure and Size on Their Trapping Efficiency in the Flow. Molecules 2022; 27:6073. [PMID: 36144805 PMCID: PMC9501256 DOI: 10.3390/molecules27186073] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/10/2022] [Revised: 09/10/2022] [Accepted: 09/13/2022] [Indexed: 11/25/2022] Open
Abstract
A promising approach to targeted drug delivery is the remote control of magnetically sensitive objects using an external magnetic field source. This method can assist in the accumulation of magnetic carriers in the affected area for local drug delivery, thus providing magnetic nanoparticles for MRI contrast and magnetic hyperthermia, as well as the magnetic separation of objects of interest from the bloodstream and liquid biopsy samples. The possibility of magnetic objects' capture in the flow is determined by the ratio of the magnetic field strength and the force of viscous resistance. Thus, the capturing ability is limited by the objects' magnetic properties, size, and flow rate. Despite the importance of a thorough investigation of this process to prove the concept of magnetically controlled drug delivery, it has not been sufficiently investigated. Here, we studied the efficiency of polyelectrolyte capsules' capture by the external magnetic field source depending on their size, the magnetic nanoparticle payload, and the suspension's flow rate. Additionally, we estimated the possibility of magnetically trapping cells containing magnetic capsules in flow and evaluated cells' membrane integrity after that. These results are required to prove the possibility of the magnetically controlled delivery of the encapsulated medicine to the affected area with its subsequent retention, as well as the capability to capture magnetically labeled cells in flow.
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Affiliation(s)
- Roman Verkhovskii
- Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia; (A.E.); (O.G.); (M.A.M.); (I.K.); (M.M.); (A.K.); (S.S.); (V.T.); (D.B.)
| | - Alexey Ermakov
- Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia; (A.E.); (O.G.); (M.A.M.); (I.K.); (M.M.); (A.K.); (S.S.); (V.T.); (D.B.)
- Institute of Molecular Theranostics, I. M. Sechenov First Moscow State Medical University, 8 Trubetskaya Str., 119991 Moscow, Russia
| | - Oleg Grishin
- Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia; (A.E.); (O.G.); (M.A.M.); (I.K.); (M.M.); (A.K.); (S.S.); (V.T.); (D.B.)
| | - Mikhail A. Makarkin
- Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia; (A.E.); (O.G.); (M.A.M.); (I.K.); (M.M.); (A.K.); (S.S.); (V.T.); (D.B.)
| | - Ilya Kozhevnikov
- Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia; (A.E.); (O.G.); (M.A.M.); (I.K.); (M.M.); (A.K.); (S.S.); (V.T.); (D.B.)
| | - Mikhail Makhortov
- Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia; (A.E.); (O.G.); (M.A.M.); (I.K.); (M.M.); (A.K.); (S.S.); (V.T.); (D.B.)
| | - Anastasiia Kozlova
- Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia; (A.E.); (O.G.); (M.A.M.); (I.K.); (M.M.); (A.K.); (S.S.); (V.T.); (D.B.)
| | - Samia Salem
- Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia; (A.E.); (O.G.); (M.A.M.); (I.K.); (M.M.); (A.K.); (S.S.); (V.T.); (D.B.)
- Department of Physics, Faculty of Science, Benha University, Benha 13511, Egypt
| | - Valery Tuchin
- Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia; (A.E.); (O.G.); (M.A.M.); (I.K.); (M.M.); (A.K.); (S.S.); (V.T.); (D.B.)
- Laboratory of Laser Molecular Imaging and Machine Learning, Tomsk State University, 36 Lenin’s Ave., 634050 Tomsk, Russia
- Institute of Precision Mechanics and Control, FRC “Saratov Scientific Centre of the Russian Academy of Sciences”, 24 Rabochaya Str., 410028 Saratov, Russia
- Bach Institute of Biochemistry, FRC “Fundamentals of Biotechnology of the Russian Academy of Sciences”, 119071 Moscow, Russia
| | - Daniil Bratashov
- Science Medical Center, Saratov State University, 83 Astrakhanskaya Str., 410012 Saratov, Russia; (A.E.); (O.G.); (M.A.M.); (I.K.); (M.M.); (A.K.); (S.S.); (V.T.); (D.B.)
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Isebia KT, Dathathri E, Verschoor N, Nanou A, De Jong AC, Coumans FAW, Terstappen LWMM, Kraan J, Martens JWM, Bansal R, Lolkema MP. Characterizing Circulating Tumor Cells and Tumor-Derived Extracellular Vesicles in Metastatic Castration-Naive and Castration-Resistant Prostate Cancer Patients. Cancers (Basel) 2022; 14:4404. [PMID: 36139564 PMCID: PMC9497200 DOI: 10.3390/cancers14184404] [Citation(s) in RCA: 7] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/30/2022] [Revised: 08/26/2022] [Accepted: 09/07/2022] [Indexed: 11/30/2022] Open
Abstract
Circulating tumor cell (CTC)- and/or tumor-derived extracellular vesicle (tdEV) loads in the blood of metastatic castration-resistant prostate cancer (CRPC) patients are associated with worse overall survival and can be used as predictive markers of treatment response. In this study, we investigated the quantity/quality of CTCs and tdEVs in metastatic castration-naive prostate cancer (CNPC) and CRPC patients, and whether androgen deprivation therapy (ADT) affects CTCs and tdEVs. We included 104 CNPC patients before ADT initiation and 66 CRPC patients. Blood samples from 31/104 CNPC patients were obtained 6 months after ADT. CTCs and tdEVs were identified using ACCEPT software. Based on the morphology, CTCs of metastatic CNPC and CRPC patients were subdivided by manual reviewing into six subclasses. The numbers of CTCs and tdEVs were correlated in both CNPC and CRPC patients, and both CTCs (p = 0.013) and tdEVs (p = 0.005) were significantly lower in CNPC compared to CRPC patients. Qualitative differences in CTCs were observed: CTC clusters (p = 0.006) and heterogeneously CK expressing CTCs (p = 0.041) were significantly lower in CNPC patients. CTC/tdEV numbers declined 6 months after ADT. Our study showed that next to CTC-load, qualitative CTC analysis and tdEV-load may be useful in CNPC patients.
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Affiliation(s)
- Khrystany T. Isebia
- Department of Medical Oncology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The Netherlands
| | - Eshwari Dathathri
- Department of Medical Cell Biophysics, Technical Medical Center, Faculty of Science and Technology, University of Twente, 7522 NB Enschede, The Netherlands
| | - Noortje Verschoor
- Department of Medical Oncology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The Netherlands
| | - Afroditi Nanou
- Department of Medical Cell Biophysics, Technical Medical Center, Faculty of Science and Technology, University of Twente, 7522 NB Enschede, The Netherlands
| | - Anouk C. De Jong
- Department of Medical Oncology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The Netherlands
| | - Frank A. W. Coumans
- Department of Medical Cell Biophysics, Technical Medical Center, Faculty of Science and Technology, University of Twente, 7522 NB Enschede, The Netherlands
| | - Leon W. M. M. Terstappen
- Department of Medical Cell Biophysics, Technical Medical Center, Faculty of Science and Technology, University of Twente, 7522 NB Enschede, The Netherlands
| | - Jaco Kraan
- Department of Medical Oncology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The Netherlands
| | - John W. M. Martens
- Department of Medical Oncology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The Netherlands
| | - Ruchi Bansal
- Department of Medical Cell Biophysics, Technical Medical Center, Faculty of Science and Technology, University of Twente, 7522 NB Enschede, The Netherlands
| | - Martijn P. Lolkema
- Department of Medical Oncology, Erasmus MC Cancer Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The Netherlands
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Stevens M, Nanou A, Terstappen LWMM, Driemel C, Stoecklein NH, Coumans FAW. StarDist Image Segmentation Improves Circulating Tumor Cell Detection. Cancers (Basel) 2022; 14:cancers14122916. [PMID: 35740582 PMCID: PMC9221404 DOI: 10.3390/cancers14122916] [Citation(s) in RCA: 25] [Impact Index Per Article: 8.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/13/2022] [Revised: 06/03/2022] [Accepted: 06/09/2022] [Indexed: 12/11/2022] Open
Abstract
Simple Summary Automated enumeration of circulating tumor cells (CTC) from immunofluorescence images starts with a selection of areas containing potential CTC. The CellSearch system has a built-in selection algorithm that has been observed to fail in samples with high cell density, thereby underestimating the true CTC load. We evaluated the deep learning method StarDist for the selection of possible CTC. In whole blood sample images, StarDist recovered 99.95% of CTC detected by CellSearch and segmented 10% additional CTC. In diagnostic leukapheresis (DLA) samples, StarDist segmented 20% additional CTC and performed well, whereas CellSearch had serious failures in 9% of samples. Abstract After a CellSearch-processed circulating tumor cell (CTC) sample is imaged, a segmentation algorithm selects nucleic acid positive (DAPI+), cytokeratin-phycoerythrin expressing (CK-PE+) events for further review by an operator. Failures in this segmentation can result in missed CTCs. The CellSearch segmentation algorithm was not designed to handle samples with high cell density, such as diagnostic leukapheresis (DLA) samples. Here, we evaluate deep-learning-based segmentation method StarDist as an alternative to the CellSearch segmentation. CellSearch image archives from 533 whole blood samples and 601 DLA samples were segmented using CellSearch and StarDist and inspected visually. In 442 blood samples from cancer patients, StarDist segmented 99.95% of CTC segmented by CellSearch, produced good outlines for 98.3% of these CTC, and segmented 10% more CTC than CellSearch. Visual inspection of the segmentations of DLA images showed that StarDist continues to perform well when the cell density is very high, whereas CellSearch failed and generated extremely large segmentations (up to 52% of the sample surface). Moreover, in a detailed examination of seven DLA samples, StarDist segmented 20% more CTC than CellSearch. Segmentation is a critical first step for CTC enumeration in dense samples and StarDist segmentation convincingly outperformed CellSearch segmentation.
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Affiliation(s)
- Michiel Stevens
- Medical Cell Biophysics Group, Techmed Center, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands; (M.S.); (A.N.); (L.W.M.M.T.)
| | - Afroditi Nanou
- Medical Cell Biophysics Group, Techmed Center, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands; (M.S.); (A.N.); (L.W.M.M.T.)
| | - Leon W. M. M. Terstappen
- Medical Cell Biophysics Group, Techmed Center, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands; (M.S.); (A.N.); (L.W.M.M.T.)
| | - Christiane Driemel
- General, Visceral and Pediatric Surgery, University Hospital and Medical Faculty, Heinrich-Heine University Düsseldorf, 40225 Düsseldorf, Germany; (C.D.); (N.H.S.)
| | - Nikolas H. Stoecklein
- General, Visceral and Pediatric Surgery, University Hospital and Medical Faculty, Heinrich-Heine University Düsseldorf, 40225 Düsseldorf, Germany; (C.D.); (N.H.S.)
| | - Frank A. W. Coumans
- Medical Cell Biophysics Group, Techmed Center, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands; (M.S.); (A.N.); (L.W.M.M.T.)
- Correspondence:
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He S, Yu S, Wei J, Ding L, Yang X, Wu Y. New horizons in the identification of circulating tumor cells (CTCs): An emerging paradigm shift in cytosensors. Biosens Bioelectron 2022; 203:114043. [PMID: 35121449 DOI: 10.1016/j.bios.2022.114043] [Citation(s) in RCA: 14] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/23/2021] [Revised: 01/02/2022] [Accepted: 01/24/2022] [Indexed: 12/11/2022]
Abstract
Circulating tumor cells (CTCs) are cancer cells that are shed from a primary tumor into the bloodstream and function as seeds for cancer metastasis at distant locations. Enrichment and identification methods of CTCs in the blood of patients plays an important role in diagnostic assessments and personalized treatments of cancer. However, the current traditional identification methods not only impact the viability of cells, but also cannot determine the type of cancer cells when the disease is unknown. Hence, new methods to identify CTCs are urgently needed. In this context, many advanced and safe technologies have emerged to distinguish between cancer cells and blood cells, and to distinguish specific types of cancer cells. In this review, at first we have briefly discussed recent advances in technologies related to the enrichment of CTCs, which lay a good foundation for the identification of CTCs. Next, we have summarized state-of-the-art technologies to confirm whether a given cell is indeed a tumor cell and determine the type of tumor cell. Finally, the challenges for application and potential directions of the current identification methods in clinical analysis of CTCs have been discussed.
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Affiliation(s)
- Sitian He
- College of Public Health, Zhengzhou University, Zhengzhou, 450001, China
| | - Songcheng Yu
- College of Public Health, Zhengzhou University, Zhengzhou, 450001, China
| | - Jinlan Wei
- College of Public Health, Zhengzhou University, Zhengzhou, 450001, China
| | - Lihua Ding
- College of Public Health, Zhengzhou University, Zhengzhou, 450001, China
| | - Xiaonan Yang
- Institute of Intelligent Sensing, Zhengzhou University, Zhengzhou, 450001, China.
| | - Yongjun Wu
- College of Public Health, Zhengzhou University, Zhengzhou, 450001, China.
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10
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Yadav A, Kumar A, Siddiqui MH. Detection of circulating tumour cells in colorectal cancer: Emerging techniques and clinical implications. World J Clin Oncol 2021; 12:1169-1181. [PMID: 35070736 PMCID: PMC8716996 DOI: 10.5306/wjco.v12.i12.1169] [Citation(s) in RCA: 13] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/01/2021] [Revised: 07/15/2021] [Accepted: 11/15/2021] [Indexed: 02/06/2023] Open
Abstract
Despite several advances in oncological management of colorectal cancer, morbidity and mortality are still high and devastating. The diagnostic evaluation by endoscopy is cumbersome, which is uncomfortable to many. Because of the intra- and inter-tumour heterogeneity and changing tumour dynamics, which is continuous in nature, the diagnostic biopsy and assessment of the pathological sample are difficult and also not adequate. Late manifestation of the disease and delayed diagnosis may lead to relapse or metastases. One of the keys to improving the outcome is early detection of cancer, ease of technology to detect with uniformity, and its therapeutic implications, which are yet to come. "Liquid biopsy" is currently the most recent area of interest in oncology, which may provide important tools regarding the characterization of the primary tumour and its metastasis as cancer cells shed into the bloodstream even at the early stages of the disease. By using this approach, clinicians may be able to find out information about the tumour at a given time. Any of the following three types of sampling of biological material can be used in the "liquid biopsy". These are circulating tumour cells (CTCs), circulating tumour DNA, and exosomes. The most commonly studied amongst the three is CTCs. CTCs with their different applications and prognostic value has been found useful in colorectal cancer detection and therapeutics. In this review, we will discuss various markers for CTCs, the core tools/techniques for detection, and also important findings of clinical studies in colorectal cancer and its clinical implications.
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Affiliation(s)
- Alka Yadav
- Department of Surgical Gastroenterology, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow 226014, Uttar Pradesh, India
| | - Ashok Kumar
- Department of Surgical Gastroenterology, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow 226014, Uttar Pradesh, India
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11
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He S, Wei J, Ding L, Yang X, Wu Y. State-of-the-arts techniques and current evolving approaches in the separation and detection of circulating tumor cell. Talanta 2021; 239:123024. [PMID: 34952370 DOI: 10.1016/j.talanta.2021.123024] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/20/2021] [Revised: 10/29/2021] [Accepted: 10/30/2021] [Indexed: 01/01/2023]
Abstract
Circulating tumor cells (CTCs) are cancer cells that shed from the primary tumor and then enter the circulatory system, a small part of which may evolve into metastatic cancer under appropriate microenvironment conditions. The detection of CTCs is a truly noninvasive, dynamic monitor for disease changes, which has considerable clinical implications in the selection of targeted drugs. However, their inherent rarity and heterogeneity pose significant challenges to their isolation and detection. Even the "gold standard", CellSearch™, suffers from high expenses, low capture efficiency, and the consumption of time. With the advancement of CTCs analysis technologies in recent years, the yield and efficiency of CTCs enrichment have gradually been improved, as well as detection sensitivity. In this review, the isolation and detection strategies of CTCs have been completely described and the potential directions for future research and development have also been highlighted through analyzing the challenges faced by current strategies.
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Affiliation(s)
- Sitian He
- College of Public Health, Zhengzhou University, Zhengzhou, 450001, China.
| | - Jinlan Wei
- College of Public Health, Zhengzhou University, Zhengzhou, 450001, China
| | - Lihua Ding
- College of Public Health, Zhengzhou University, Zhengzhou, 450001, China
| | - Xiaonan Yang
- School of Information Engineering, Zhengzhou University, Zhengzhou, 450001, China.
| | - Yongjun Wu
- College of Public Health, Zhengzhou University, Zhengzhou, 450001, China.
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12
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Huang C, Lin X, He J, Liu N. Enrichment and detection method for the prognostic value of circulating tumor cells in ovarian cancer: A meta-analysis. Gynecol Oncol 2021; 161:613-620. [PMID: 33674144 DOI: 10.1016/j.ygyno.2021.02.024] [Citation(s) in RCA: 18] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/21/2020] [Accepted: 02/17/2021] [Indexed: 02/07/2023]
Abstract
OBJECTIVES Recent studies have revealed that circulating tumor cells (CTCs) might predict bad prognosis, but the results were conflicting. Sampling time, treatment, enrichment method and detection method also varied. We aimed to evaluate whether patients with CTCs in peripheral blood have bad survival outcomes with consideration of the above four aspects. METHODS Relevant studies were searched on Pubmed, Embase and the Cochrane Library. Studies of CTCs involving survival data available were identified for a systematic review and meta-analysis. HRs and 95% CIs for PFS and OS were extracted directly or from the Kaplan-Meier survival curves by the Engauge Digitizer v4.1. Subgroup analyses were performed to evaluate the effect of sampling time, treatment, enrichment method and detection method. RESULTS Two clinical trials and thirteen retrospective studies with a total of 1285 patients were included. CTCs significantly correlated with OS (HR = 1.77, 95%CI:1.42-2.21, p < 0.00001 and PFS (HR = 1.53, 95%CI:1.26-1.86, p < 0.0001). Subgroup analyses showed that CTCs were significant associated with OS in the "Pre-therapy" subgroup (HR = 1.79, 95%CI:1.43-2.24, p < 0.00001), the "Surgery" group (HR = 1.82, 95%CI:1.42-2.33, p < 0.00001), and the "RT-PCR"subgroup (HR = 2.29, 95%CI:1.53-3.42, p < 0.0001). While for enrichment method, CTCs significantly correlated with OS in the"Physical method" subgroup (HR = 1.94, 95%CI:1.21-3.09, p = 0.006) and the "Immunological method" subgroup (HR = 1.84, 95%CI:1.37-2.48, p < 0.0001). CONCLUSIONS The presence of CTCs prior to the treatment indicated worse OS and PFS and CTCs might be predictive biomarker for ovarian cancer patients . CTCs detected using RT-PCR seem to be associated with poorer OS and PFS in patients with ovarian cancer.
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Affiliation(s)
- Chengying Huang
- Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou, China
| | - Xiaoli Lin
- Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou, China
| | - Jinmei He
- Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou, China
| | - Nan Liu
- Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
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Nanou A, Mol L, Coumans FAW, Koopman M, Punt CJA, Terstappen LWMM. Endothelium-Derived Extracellular Vesicles Associate with Poor Prognosis in Metastatic Colorectal Cancer. Cells 2020; 9:E2688. [PMID: 33333805 PMCID: PMC7765205 DOI: 10.3390/cells9122688] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/23/2020] [Revised: 12/01/2020] [Accepted: 12/11/2020] [Indexed: 12/14/2022] Open
Abstract
Elevated, tumor-derived extracellular vesicle (tdEV) and circulating tumor cell (CTC) loads in metastatic cancer are associated with poor clinical outcome. Herein, we investigate whether endothelium-derived extracellular vesicles (edEVs) can be detected in the blood of metastatic colorectal cancer (mCRC) patients, and whether those vesicles associate with prognosis. The open-source ACCEPT (Automated CTC Classification, Enumeration, and Phenotyping) software was used to enumerate edEVs, tdEVs, and other objects from digitally stored CellSearch images acquired after CTC and circulating endothelial cell (CEC) enrichment from the blood of 395 mCRC patients before the initiation of a new therapy. Patients had participated in the prospective phase III CAIRO2 study. The presence of edEVs was found 5- to 10-fold higher than CECs. The hazard ratio (HR) (95% CI) of progression-free survival (PFS) for increased CTCs (≥3 in 7.5 mL), tdEVs (≥40 in 7.5 mL), and edEVs (≥287 in 4.0 mL.) was 1.4 (1.1-1.9), 2.0 (1.5-2.6), and 1.7 (1.2-2.5), respectively. The HR of Overall Survival (OS) for increased CTCs, tdEVs and edEVs was 2.2 (1.7-3.0), 2.7 (2.0-3.5), and 2.1 (1.5-2.8), respectively. There was no cut-off value for CECs, leading to a dichotomization of patients with a significant HR. Only tdEVs remained a significant predictor of OS in the final multivariable model.
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Affiliation(s)
- Afroditi Nanou
- Department of Medical Cell BioPhysics, University of Twente, 7522ND Enschede, The Netherlands;
| | - Linda Mol
- Netherlands Comprehensive Cancer Organization, 6533AA Nijmegen, The Netherlands;
| | - Frank A. W. Coumans
- Department of Medical Cell BioPhysics, University of Twente, 7522ND Enschede, The Netherlands;
| | - Miriam Koopman
- Department of Medical Oncology, University Medical Center Utrecht, 3584CS Utrecht, The Netherlands;
| | - Cornelis J. A. Punt
- Julius Center for Health Sciences and Primary Care, Department of Epidemiology, University Medical Center Utrecht, 3584CG Utrecht, The Netherlands;
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14
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Liu P, Jonkheijm P, Terstappen LWMM, Stevens M. Magnetic Particles for CTC Enrichment. Cancers (Basel) 2020; 12:cancers12123525. [PMID: 33255978 PMCID: PMC7760229 DOI: 10.3390/cancers12123525] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/21/2020] [Revised: 11/20/2020] [Accepted: 11/23/2020] [Indexed: 02/07/2023] Open
Abstract
Simple Summary For the enrichment of very rare cells, such as Circulating Tumor Cells (CTCs), immunomagnetic enrichment is frequently used. For this purpose, magnetic nanoparticles (MNPs) coated with specific antibodies directed against cancer cells are used. In this review, we look at the properties such a particle needs to have in order to be used successfully, and describe the different methods used in the production of such a particle as well as the methods for their separation. Additionally, an overview is given of the antibodies that could potentially be used for this purpose. Abstract Here, we review the characteristics and synthesis of magnetic nanoparticles (MNPs) and place these in the context of their usage in the immunomagnetic enrichment of Circulating Tumor Cells (CTCs). The importance of the different characteristics is explained, the need for a very specific enrichment is emphasized and different (commercial) magnetic separation techniques are shown. As the specificity of an MNP is in a large part dependent on the antibody coated onto the particle, different strategies in the coupling of specific antibodies as well as an overview of the available antibodies is given.
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Affiliation(s)
- Peng Liu
- Department of Medical Cell BioPhysics, University of Twente, 7522 NB Enschede, The Netherlnds; (P.L.); (L.W.M.M.T.)
- Department of Molecular Nanofabrication, University of Twente, 7522 NB Enschede, The Netherlands;
| | - Pascal Jonkheijm
- Department of Molecular Nanofabrication, University of Twente, 7522 NB Enschede, The Netherlands;
| | - Leon W. M. M. Terstappen
- Department of Medical Cell BioPhysics, University of Twente, 7522 NB Enschede, The Netherlnds; (P.L.); (L.W.M.M.T.)
| | - Michiel Stevens
- Department of Medical Cell BioPhysics, University of Twente, 7522 NB Enschede, The Netherlnds; (P.L.); (L.W.M.M.T.)
- Correspondence: ; Tel.: +31-53-489-4101
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Wang SG, Zhang B, Li CG, Zhu JQ, Sun BS, Wang CL. Sorting and gene mutation verification of circulating tumor cells of lung cancer with epidermal growth factor receptor peptide lipid magnetic spheres. Thorac Cancer 2020; 11:2887-2895. [PMID: 32856417 PMCID: PMC7529546 DOI: 10.1111/1759-7714.13625] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/08/2020] [Revised: 07/29/2020] [Accepted: 07/30/2020] [Indexed: 12/26/2022] Open
Abstract
Background This study aimed to identify an efficient, simple, and specific method of detecting mutations in the epidermal growth factor receptor (EGFR) gene in isolated lung cancer circulating tumor cells (CTCs) and to improve the ability to obtain tumor tissue clinically. Methods EGFR peptide lipid magnetic spheres (EG‐P‐LMB) were prepared by reverse evaporation, and characterization and cell capture efficiency assessed. The peripheral blood samples of 30 lung cancer patients were isolated and identified with the EG‐P‐LMB using 20 healthy volunteers as controls. Finally, the isolated CTCs were tested for EGFR gene mutations, and the tissue samples selected for comparison. Results The prepared magnetic spheres had a smaller particle size and higher stability according to the particle size potential test. Their morphology was homogeneous by atomic force observation, and the UV test showed that there were peptides on the surface. The separation efficiency of EG‐P‐LMB was greater than 90% in PBS and greater than 80% in the blood simulation system. Compared with the tissue sample results, the positive rate of EGFR gene mutations was 94%. The CTC test results of 27 patients were consistent with the tissue test results of the corresponding patients, and the consistency with the tissue comparison test results was 90% (27/30). Conclusions EG‐P‐LMB can effectively capture CTCs in the peripheral blood of patients with lung cancer. CTC detection can accurately identify mutations in the EGFR gene and improve the ability to obtain tumor tissue in clinical practice. Key points Significant findings of the study EG‐P‐LMB can effectively capture CTCs in the peripheral blood of patients with lung cancer. CTC detection can accurately identify mutations in the EGFR gene and improve the ability to obtain tumor tissue in clinical practice. What this study adds This study added EGFR peptide lipid magnetic spheres to capture CTCs in the blood. Genetic testing was performed and compared with tissues. It solves the problem of clinically difficult tumor tissue sampling.
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Affiliation(s)
- Sheng-Guang Wang
- Department of Lung Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin Lung Cancer Center, Tianjin, China
| | - Bin Zhang
- Department of Lung Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin Lung Cancer Center, Tianjin, China
| | - Chen-Guang Li
- Department of Lung Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin Lung Cancer Center, Tianjin, China
| | - Jian-Quan Zhu
- Department of Lung Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin Lung Cancer Center, Tianjin, China
| | - Bing-Sheng Sun
- Department of Lung Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin Lung Cancer Center, Tianjin, China
| | - Chang-Li Wang
- Department of Lung Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin's Clinical Research Center for Cancer, Tianjin Lung Cancer Center, Tianjin, China
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16
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Ahrens TD, Bang-Christensen SR, Jørgensen AM, Løppke C, Spliid CB, Sand NT, Clausen TM, Salanti A, Agerbæk MØ. The Role of Proteoglycans in Cancer Metastasis and Circulating Tumor Cell Analysis. Front Cell Dev Biol 2020; 8:749. [PMID: 32984308 PMCID: PMC7479181 DOI: 10.3389/fcell.2020.00749] [Citation(s) in RCA: 57] [Impact Index Per Article: 11.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/07/2020] [Accepted: 07/17/2020] [Indexed: 12/14/2022] Open
Abstract
Circulating tumor cells (CTCs) are accessible by liquid biopsies via an easy blood draw. They represent not only the primary tumor site, but also potential metastatic lesions, and could thus be an attractive supplement for cancer diagnostics. However, the analysis of rare CTCs in billions of normal blood cells is still technically challenging and novel specific CTC markers are needed. The formation of metastasis is a complex process supported by numerous molecular alterations, and thus novel CTC markers might be found by focusing on this process. One example of this is specific changes in the cancer cell glycocalyx, which is a network on the cell surface composed of carbohydrate structures. Proteoglycans are important glycocalyx components and consist of a protein core and covalently attached long glycosaminoglycan chains. A few CTC assays have already utilized proteoglycans for both enrichment and analysis of CTCs. Nonetheless, the biological function of proteoglycans on clinical CTCs has not been studied in detail so far. Therefore, the present review describes proteoglycan functions during the metastatic cascade to highlight their importance to CTCs. We also outline current approaches for CTC assays based on targeting proteoglycans by their protein cores or their glycosaminoglycan chains. Lastly, we briefly discuss important technical aspects, which should be considered for studying proteoglycans.
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Affiliation(s)
- Theresa D. Ahrens
- Centre for Medical Parasitology at Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, Copenhagen, Denmark
| | - Sara R. Bang-Christensen
- Centre for Medical Parasitology at Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, Copenhagen, Denmark
- VarCT Diagnostics, Copenhagen, Denmark
| | | | - Caroline Løppke
- Centre for Medical Parasitology at Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, Copenhagen, Denmark
| | - Charlotte B. Spliid
- Centre for Medical Parasitology at Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, Copenhagen, Denmark
- Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA, United States
| | - Nicolai T. Sand
- Centre for Medical Parasitology at Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, Copenhagen, Denmark
| | - Thomas M. Clausen
- Centre for Medical Parasitology at Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, Copenhagen, Denmark
- Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA, United States
| | - Ali Salanti
- Centre for Medical Parasitology at Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, Copenhagen, Denmark
| | - Mette Ø. Agerbæk
- Centre for Medical Parasitology at Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, Copenhagen, Denmark
- VarCT Diagnostics, Copenhagen, Denmark
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Tumour-derived extracellular vesicles in blood of metastatic cancer patients associate with overall survival. Br J Cancer 2020; 122:801-811. [PMID: 31937922 PMCID: PMC7078322 DOI: 10.1038/s41416-019-0726-9] [Citation(s) in RCA: 58] [Impact Index Per Article: 11.6] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/19/2019] [Revised: 12/17/2019] [Accepted: 12/19/2019] [Indexed: 12/20/2022] Open
Abstract
BACKGROUND Circulating tumour cells (CTCs) in blood associate with overall survival (OS) of cancer patients, but they are detected in extremely low numbers. Large tumour-derived extracellular vesicles (tdEVs) in castration-resistant prostate cancer (CRPC) patients are present at around 20 times higher frequencies than CTCs and have equivalent prognostic power. In this study, we explored the presence of tdEVs in other cancers and their association with OS. METHODS The open-source ACCEPT software was used to automatically enumerate tdEVs in digitally stored CellSearch® images obtained from previously reported CTC studies evaluating OS in 190 CRPC, 450 metastatic colorectal cancer (mCRC), 179 metastatic breast cancer (MBC) and 137 non-small cell lung cancer (NSCLC) patients before the initiation of a new treatment. RESULTS Presence of unfavourable CTCs and tdEVs is predictive of OS, with respective hazard ratios (HRs) of 2.4 and 2.2 in CRPC, 2.7 and 2.2 in MBC, 2.3 and 1.9 in mCRC and 2.0 and 2.4 in NSCLC, respectively. CONCLUSIONS tdEVs have equivalent prognostic value as CTCs in the investigated metastatic cancers. CRPC, mCRC, and MBC (but not NSCLC) patients with favourable CTC counts can be further prognostically stratified using tdEVs. Our data suggest that tdEVs could be used in clinical decision-making.
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18
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Yousefi M, Ghaffari P, Nosrati R, Dehghani S, Salmaninejad A, Abarghan YJ, Ghaffari SH. Prognostic and therapeutic significance of circulating tumor cells in patients with lung cancer. Cell Oncol (Dordr) 2019; 43:31-49. [PMID: 31828552 DOI: 10.1007/s13402-019-00470-y] [Citation(s) in RCA: 38] [Impact Index Per Article: 6.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 08/06/2019] [Indexed: 12/11/2022] Open
Abstract
BACKGROUND Lung cancer is the second most common cancer and the main cause of cancer-related mortality worldwide. In spite of various efforts that have been made to facilitate the early diagnosis of lung cancer, most patients are diagnosed when the disease is already in stage IV, which is generally associated with the occurrence of distant metastases and a poor survival. Moreover, a large proportion of these patients will relapse after treatment, heralding the need for the stratification of lung cancer patients in addition to identifying those who are at a higher risk of relapse and, thus, require alternative and/or additional therapies. Recently, circulating tumor cells (CTCs) have been considered as valuable markers for the early diagnosis, prognosis and risk stratification of cancer patients, and they have been found to be able to predict the survival of patients with various types of cancer, including lung cancer. Additionally, the characterization of CTCs has recently provided fascinating insights into the heterogeneity of tumors, which may be instrumental for the development of novel targeted therapies. CONCLUSIONS Here we review our current understanding of the significance of CTCs in lung cancer metastasis. We also discuss prominent studies reporting the utility of enumeration and characterization of CTCs in lung cancer patients as prognostic and pharmacodynamic biomarkers for those who are at a higher risk of metastasis and drug resistance.
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Affiliation(s)
- Meysam Yousefi
- Department of Medical Genetics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.,Department of Medical Genetics, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Parisa Ghaffari
- Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran
| | - Rahim Nosrati
- Cellular and Molecular Research Center, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran.,Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran
| | - Sadegh Dehghani
- Department of Medical Biotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
| | - Arash Salmaninejad
- Department of Medical Genetics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.,Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
| | - Yousef Jafari Abarghan
- Department of Medical Genetics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
| | - Seyed H Ghaffari
- Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran.
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EpCAM low Circulating Tumor Cells: Gold in the Waste. DISEASE MARKERS 2019; 2019:1718920. [PMID: 31636732 PMCID: PMC6766153 DOI: 10.1155/2019/1718920] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Subscribe] [Scholar Register] [Received: 05/03/2019] [Revised: 09/06/2019] [Accepted: 09/11/2019] [Indexed: 12/14/2022]
Abstract
The CellSearch® system which is still considered the gold standard for the enumeration of circulating tumor cells (CTC) utilizes antibodies against the epithelial cell adhesion molecule (EpCAM) for CTC enrichment. Recently, CTC discarded by the CellSearch® system due to their low EpCAM expression have been isolated and analyzed. We here sought to discuss technical and biological issues concerning the isolation and characterization of EpCAMlow CTC, highlighting the enormous potential of this subpopulation discarded by CellSearch®, which might instead reveal an unexpected clinical significance in tumor types where CTC enumeration has never been validated for prognostic and predictive purpose.
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20
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Leukocyte-Derived Extracellular Vesicles in Blood with and without EpCAM Enrichment. Cells 2019; 8:cells8080937. [PMID: 31434250 PMCID: PMC6721753 DOI: 10.3390/cells8080937] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/29/2019] [Revised: 08/07/2019] [Accepted: 08/15/2019] [Indexed: 12/23/2022] Open
Abstract
Large tumor-derived Extracellular Vesicles (tdEVs) detected in blood of metastatic prostate, breast, colorectal, and non-small cell lung cancer patients after enrichment for Epithelial Cell Adhesion Molecule (EpCAM) expression and labeling with 4',6-diamidino-2-phenylindole (DAPI), phycoerythrin-conjugated antibodies against Cytokeratins (CK-PE), and allophycocyanin-conjugated antibody against the cluster of differentiation 45 (CD45-APC), are negatively associated with the overall survival of patients. Here, we investigated whether, similarly to tdEVs, leukocyte-derived EVs (ldEVs) could also be detected in EpCAM-enriched blood. Presence of ldEVs and leukocytes in image data sets of EpCAM-enriched samples of 25 healthy individuals and 75 metastatic cancer patients was evaluated using the ACCEPT software. Large ldEVs could indeed be detected, but in contrast to the 20-fold higher frequency of tdEVs as compared to Circulating Tumor Cells (CTCs), ldEVs were present in a 5-fold lower frequency as compared to leukocytes. To evaluate whether these ldEVs pre-exist in the blood or are formed during the CellSearch procedure, the blood of healthy individuals without EpCAM enrichment was labelled with the nuclear dye Hoechst and fluorescently tagged monoclonal antibodies recognizing the leukocyte-specific CD45, platelet-specific CD61, and red blood cell-specific CD235a. Fluorescence microscopy imaging using a similar setup as the CellSearch was performed and demonstrated the presence of a similar population of ldEVs present at a 3-fold lower frequency as compared to leukocytes.
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21
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Beck TN, Boumber YA, Aggarwal C, Pei J, Thrash-Bingham C, Fittipaldi P, Vlasenkova R, Rao C, Borghaei H, Cristofanilli M, Mehra R, Serebriiskii I, Alpaugh RK. Circulating tumor cell and cell-free RNA capture and expression analysis identify platelet-associated genes in metastatic lung cancer. BMC Cancer 2019; 19:603. [PMID: 31215484 PMCID: PMC6582501 DOI: 10.1186/s12885-019-5795-x] [Citation(s) in RCA: 29] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/11/2018] [Accepted: 06/05/2019] [Indexed: 12/25/2022] Open
Abstract
Background Circulating tumor cells (CTC) and plasma cell-free RNA (cfRNA) can serve as biomarkers for prognosis and treatment response in lung cancer. One barrier to the selected or routine use of CTCs and plasma cfRNA in precision oncology is the limited quantity of both, and CTCs are only seen in metastatic disease. As capture of CTCs and plasma cfRNA presents an opportunity to monitor and assess malignancies without invasive procedures, we compared two methods for CTC capture and identification, and profiled mRNA from CTCs and plasma cfRNA to identify potential tumor-associated biomarkers. Methods Peripheral blood was collected from ten patients with small cell lung cancer (SCLC), ten patients with non-small cell lung cancer (NSCLC) and four healthy volunteers. Two methods were used for CTC capture: the standard epithelial cell adhesion molecule (EpCam) CellSearch kit (unicapture) and EpCAM plus HER2, EGFR and MUC-1 specific combined ferrofluid capture (quadcapture). For the quadcapture, anti-cytokeratin 7 (CK7) was additionally used to assist in CTC identification. NanoString analysis was performed on plasma cfRNA and on mRNA from combined ferrofluid isolated CTCs. Expression data was analyzed using STRING and Reactome. Results Unicapture detected CTCs in 40% of NSCLC and 60% of SCLC; whereas, quadcapture/CK7 identified CTCs in 20% of NSCLC and 80% of SCLC. Bioinformatic analysis of NanoString data identified high expression of a platelet factor 4 (PF4)-related group of transcripts. Conclusions Quadcapture ferrofluid reagent did not significantly improve CTC capture efficacy. NanoString analysis based on CTC and plasma cfRNA data highlighted an intriguing PF-4-centric network in patients with metastatic lung cancer. Electronic supplementary material The online version of this article (10.1186/s12885-019-5795-x) contains supplementary material, which is available to authorized users.
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Affiliation(s)
- Tim N Beck
- Program in Molecular Therapeutics, Fox Chase Cancer Center, Philadelphia, PA, 19111, USA.,Digestive Disease & Surgery Institute, Cleveland Clinic, Cleveland, OH, 44195, USA
| | - Yanis A Boumber
- Program in Molecular Therapeutics, Fox Chase Cancer Center, Philadelphia, PA, 19111, USA.,Department of Hematology/Oncology, Fox Chase Cancer Center, Philadelphia, PA, 19111, USA.,Kazan Federal University, Kazan, Russian Federation
| | - Charu Aggarwal
- Abramson Cancer Center and Division of Hematology/Oncology, Department of Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, 19104, USA
| | - Jianming Pei
- Genomics Facility, Fox Chase Cancer Center, Philadelphia, PA, 19111, USA
| | | | - Patricia Fittipaldi
- Protocol Support Laboratory, Fox Chase Cancer Center, Philadelphia, PA, 19111, USA
| | | | - Chandra Rao
- Janssen Diagnostics LLC, Valley, Huntingdon, PA, 19006, USA
| | - Hossein Borghaei
- Program in Molecular Therapeutics, Fox Chase Cancer Center, Philadelphia, PA, 19111, USA.,Department of Hematology/Oncology, Fox Chase Cancer Center, Philadelphia, PA, 19111, USA
| | - Massimo Cristofanilli
- Feinberg School of Medicine, Robert H Lurie Comprehensive Cancer Center, Chicago, IL, 60611, USA
| | - Ranee Mehra
- Head and Neck Medical Oncology, University of Maryland Greenebaum Comprehensive Cancer Center, Baltimore, MD, 21201, USA
| | - Ilya Serebriiskii
- Program in Molecular Therapeutics, Fox Chase Cancer Center, Philadelphia, PA, 19111, USA.,Kazan Federal University, Kazan, Russian Federation
| | - R Katherine Alpaugh
- Protocol Support Laboratory, Fox Chase Cancer Center, Philadelphia, PA, 19111, USA. .,Biostatistics Facility, Fox Chase Cancer Center, Philadelphia, PA, 19111, USA.
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22
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Nicolazzo C, Busetto GM, Gradilone A, Sperduti I, Del Giudice F, Loreni F, Cortesi E, de Berardinis E, Gazzaniga P, Raimondi C. Circulating Tumor Cells Identify Patients with Super-High-Risk Non-Muscle-Invasive Bladder Cancer: Updated Outcome Analysis of a Prospective Single-Center Trial. Oncologist 2019; 24:612-616. [PMID: 30944184 DOI: 10.1634/theoncologist.2018-0784] [Citation(s) in RCA: 33] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/15/2018] [Accepted: 01/31/2019] [Indexed: 11/17/2022] Open
Abstract
BACKGROUND Clinical behavior of non-muscle-invasive bladder cancer (NMIBC) is largely unpredictable, and even patients treated according to European Association of Urology recommendations have a heterogeneous prognosis. High-grade T1 (HGT1) bladder cancer is the highest-risk subtype of NMIBC, with an almost 40% rate of recurrence and 20% of progression at 5 years. Nomograms predicting risk of recurrence, progression, and cancer-specific survival (CSS) are not available specifically within HGT1 bladder cancer, and the identification of robust prognostic biomarkers to better guide therapeutic strategies in this subgroup of patients is of paramount importance. Strategies to identify putative biomarkers in liquid biopsies from blood and urine collected from patients with bladder cancer have been intensively studied in the last few years. SUBJECTS, MATERIALS, AND METHODS We here report the final analysis of a single-center prospective study aimed to investigate the impact of circulating tumor cells (CTCs) on CSS and overall survival (OS) in 102 patients with HGT1 bladder cancer, in a median follow-up of 63 months. RESULTS We here demonstrate that the presence of even a single CTC is predictive of shorter CSS and OS, as compared with the standard predictive variables. Points of attention in this multivariable analysis are the long-term follow-up and the adequate number of outcome events. CONCLUSION The accurate risk stratification provided by CTCs might be essential for determining the best surveillance strategy for patients after diagnosis. A closer follow-up, an early radical surgery, or even a systemic treatment might be recommended in patients with super-high-risk non-muscle-invasive bladder cancer. IMPLICATIONS FOR PRACTICE Circulating tumor cells identify patients with super-high-risk non-muscle-invasive bladder cancer who require closer monitoring for local recurrence and/or progression of disease. This super-high-risk subgroup of patients might also require more aggressive treatment interventions, which should be evaluated in large prospective cohorts.
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Affiliation(s)
- Chiara Nicolazzo
- Dipartimento Medicina Molecolare, Sapienza Università di Roma, Rome, Italy
| | - Gian Maria Busetto
- Dipartimento Scienze Ginecologico-Ostetriche e Scienze Urologiche, Sapienza Università di Roma, Rome, Italy
| | - Angela Gradilone
- Dipartimento Medicina Molecolare, Sapienza Università di Roma, Rome, Italy
| | | | - Francesco Del Giudice
- Dipartimento Scienze Ginecologico-Ostetriche e Scienze Urologiche, Sapienza Università di Roma, Rome, Italy
| | - Flavia Loreni
- Dipartimento Medicina Molecolare, Sapienza Università di Roma, Rome, Italy
| | - Enrico Cortesi
- Dipartimento Scienze Radiologiche, Oncologiche e Patologiche, Sapienza Università di Roma, Rome, Italy
| | - Ettore de Berardinis
- Dipartimento Scienze Ginecologico-Ostetriche e Scienze Urologiche, Sapienza Università di Roma, Rome, Italy
| | - Paola Gazzaniga
- Dipartimento Medicina Molecolare, Sapienza Università di Roma, Rome, Italy
| | - Cristina Raimondi
- Dipartimento Scienze Radiologiche, Oncologiche e Patologiche, Sapienza Università di Roma, Rome, Italy
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23
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Rostami P, Kashaninejad N, Moshksayan K, Saidi MS, Firoozabadi B, Nguyen NT. Novel approaches in cancer management with circulating tumor cell clusters. JOURNAL OF SCIENCE: ADVANCED MATERIALS AND DEVICES 2019; 4:1-18. [DOI: 10.1016/j.jsamd.2019.01.006] [Citation(s) in RCA: 36] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/06/2025]
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24
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Baird Z, Cao Z, Barron MR, Vorsilak A, Deiss F, Pugia M. Enumeration of Rare Cells in Whole Blood by Signal Ion Emission Reactive Release Amplification with Same-Sample RNA Analysis. Anal Chem 2019; 91:2028-2034. [DOI: 10.1021/acs.analchem.8b04446] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
Affiliation(s)
- Zane Baird
- Single Cell Analytics Center, Indiana Biosciences Research Institute, Indianapolis, Indiana 46202, United States
| | - Zehui Cao
- Single Cell Analytics Center, Indiana Biosciences Research Institute, Indianapolis, Indiana 46202, United States
| | - M. Regina Barron
- Single Cell Analytics Center, Indiana Biosciences Research Institute, Indianapolis, Indiana 46202, United States
- Department of Chemistry and Chemical Biology, Indiana University−Purdue University Indianapolis, Indianapolis, Indiana 46202, United States
| | - Anna Vorsilak
- Single Cell Analytics Center, Indiana Biosciences Research Institute, Indianapolis, Indiana 46202, United States
| | - Frédérique Deiss
- Department of Chemistry and Chemical Biology, Indiana University−Purdue University Indianapolis, Indianapolis, Indiana 46202, United States
| | - Michael Pugia
- Single Cell Analytics Center, Indiana Biosciences Research Institute, Indianapolis, Indiana 46202, United States
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25
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Nicolazzo C, Gradilone A, Carpino G, Gazzaniga P, Raimondi C. Molecular Characterization of Circulating Tumor Cells to Study Cancer Immunoevasion. Methods Mol Biol 2019; 1884:247-258. [PMID: 30465208 DOI: 10.1007/978-1-4939-8885-3_17] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 06/09/2023]
Abstract
Cancer cells leaving the primary tumor immunosuppressive microenvironment become vulnerable to active immune surveillance and require mechanisms of immunoevasion to survive in the circulation. Studies have identified several pathways by which circulating tumor cells (CTCs) might escape the immune system/immunotherapy attack. The PD-1/PD-L1 axis is an immune checkpoint regulator, playing a major role in maintaining self-tolerance. It is now well recognized that tumor cells co-opt the PD-1/PD-L1 axis of immune regulation to interfere with cytotoxic T lymphocyte function. Transcriptional changes in CTCs, leading to the upregulation of PD-L1, might enable them to survive in circulation. Very recent data revealed a previously unappreciated role of epithelial-mesenchymal transition (EMT) in reprogramming the immune response in the local tumor microenvironment and a mutual regulation between EMT and immunoevasion is becoming apparent. In this chapter, we will describe in detail both EpCAM-dependent and -independent approaches that allow the identification of PD-L1 expression and EMT-like features in circulating tumor cells.
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Affiliation(s)
- Chiara Nicolazzo
- Dipartimento di Medicina Molecolare, Sapienza Università di Roma, Rome, Italy
| | - Angela Gradilone
- Dipartimento di Medicina Molecolare, Sapienza Università di Roma, Rome, Italy
| | - Guido Carpino
- Dipartimento di Anatomia, Istologia, Medicina Forense e Scienze Ortopediche, Sapienza Università di Roma, Rome, Italy
| | - Paola Gazzaniga
- Dipartimento di Medicina Molecolare, Sapienza Università di Roma, Rome, Italy
| | - Cristina Raimondi
- Dipartimento di Medicina Molecolare, Sapienza Università di Roma, Rome, Italy.
- Dipartimento di Scienze Radiologiche, Oncologiche ed Anatomopatologiche, Sapienza Università di Roma, Rome, Italy.
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26
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Breuninger S, Stangl S, Werner C, Sievert W, Lobinger D, Foulds GA, Wagner S, Pickhard A, Piontek G, Kokowski K, Pockley AG, Multhoff G. Membrane Hsp70-A Novel Target for the Isolation of Circulating Tumor Cells After Epithelial-to-Mesenchymal Transition. Front Oncol 2018; 8:497. [PMID: 30443493 PMCID: PMC6223102 DOI: 10.3389/fonc.2018.00497] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/09/2018] [Accepted: 10/12/2018] [Indexed: 12/29/2022] Open
Abstract
The presence of circulating tumor cells (CTCs) in the peripheral blood is a pre-requisite for progression, invasion, and metastatic spread of cancer. Consequently, the enumeration and molecular characterization of CTCs from the peripheral blood of patients with solid tumors before, during and after treatment serves as a valuable tool for categorizing disease, evaluating prognosis and for predicting and monitoring therapeutic responsiveness. Many of the techniques for isolating CTCs are based on the expression of epithelial cell surface adhesion molecule (EpCAM, CD326) on tumor cells. However, the transition of adherent epithelial cells to migratory mesenchymal cells (epithelial-to-mesenchymal transition, EMT)-an essential element of the metastatic process-is frequently associated with a loss of expression of epithelial cell markers, including EpCAM. A highly relevant proportion of mesenchymal CTCs cannot therefore be isolated using techniques that are based on the "capture" of cells expressing EpCAM. Herein, we provide evidence that a monoclonal antibody (mAb) directed against a membrane-bound form of Hsp70 (mHsp70)-cmHsp70.1-can be used for the isolation of viable CTCs from peripheral blood of tumor patients of different entities in a more quantitative manner. In contrast to EpCAM, the expression of mHsp70 remains stably upregulated on migratory, mesenchymal CTCs, metastases and cells that have been triggered to undergo EMT. Therefore, we propose that approaches for isolating CTCs based on the capture of cells that express mHsp70 using the cmHsp70.1 mAb are superior to those based on EpCAM expression.
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Affiliation(s)
- Stephanie Breuninger
- Center for Translational Cancer Research TU München (TranslaTUM), Klinikum rechts der Isar, TUM, Munich, Germany
| | - Stefan Stangl
- Center for Translational Cancer Research TU München (TranslaTUM), Klinikum rechts der Isar, TUM, Munich, Germany
| | - Caroline Werner
- Center for Translational Cancer Research TU München (TranslaTUM), Klinikum rechts der Isar, TUM, Munich, Germany
| | - Wolfgang Sievert
- Center for Translational Cancer Research TU München (TranslaTUM), Klinikum rechts der Isar, TUM, Munich, Germany
| | - Dominik Lobinger
- Center for Translational Cancer Research TU München (TranslaTUM), Klinikum rechts der Isar, TUM, Munich, Germany
| | - Gemma A Foulds
- John van Geest Cancer Research Centre, College of Science and Technology, Nottingham Trent University, Nottingham, United Kingdom
| | - Sarah Wagner
- John van Geest Cancer Research Centre, College of Science and Technology, Nottingham Trent University, Nottingham, United Kingdom
| | - Anja Pickhard
- Department of Otolaryngology Head and Neck Surgery, Klinikum rechts der Isar, TUM, Munich, Germany
| | - Guido Piontek
- Department of Otolaryngology Head and Neck Surgery, Klinikum rechts der Isar, TUM, Munich, Germany
| | - Konrad Kokowski
- Department of Pneumology and Pneumologic Oncology, Klinikum Bogenhausen, Munich, Germany
| | - Alan G Pockley
- John van Geest Cancer Research Centre, College of Science and Technology, Nottingham Trent University, Nottingham, United Kingdom
| | - Gabriele Multhoff
- Center for Translational Cancer Research TU München (TranslaTUM), Klinikum rechts der Isar, TUM, Munich, Germany
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27
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Song Z, Liu Y. [Progress of Liquid Biopsy in Early Diagnosis of Lung Cancer]. ZHONGGUO FEI AI ZA ZHI = CHINESE JOURNAL OF LUNG CANCER 2018; 21:620-627. [PMID: 30172270 PMCID: PMC6105353 DOI: 10.3779/j.issn.1009-3419.2018.08.08] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
肺癌的早期诊断有利于提高患者的生存率。应用影像学方法对肺癌高风险人群进行筛查，可以起到早发现、早诊断的作用。越来越多的研究显示，液体活检（liquid biopsy）可以对该方法进行替代和补充。检测肺癌患者外周血中的循环肿瘤细胞（circulating tumor cells, CTCs）、循环肿瘤DNA（circulating tumor DNA, ctDNA）、微小核糖核酸（microRNA, miRNA）、外泌体（exosomes）、肿瘤血小板（tumor educated platelets, TEPs）可以用于肺癌的早期诊断，并且可能为影像学检查阴性的高风险人群提供相应的诊疗建议。全文就以上标志物的检测手段、在肺癌早期诊断中的价值以及存在优势与局限性进行综述，以期促进液体活检在肺癌早期诊断、与其他筛查手段相结合方面的应用。
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Affiliation(s)
- Zhipeng Song
- Department of Epidemiology, Beijing Chest Hospital, Capital Medical University, Beijing 101149, China
| | - Yang Liu
- Department of Epidemiology, Beijing Chest Hospital, Capital Medical University, Beijing 101149, China
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28
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de Wit S, Zeune LL, Hiltermann TJN, Groen HJM, Dalum GV, Terstappen LWMM. Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis. Cancers (Basel) 2018; 10:cancers10100377. [PMID: 30308977 PMCID: PMC6210778 DOI: 10.3390/cancers10100377] [Citation(s) in RCA: 28] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/21/2018] [Revised: 10/05/2018] [Accepted: 10/08/2018] [Indexed: 02/05/2023] Open
Abstract
In the CellSearch® system, blood is immunomagnetically enriched for epithelial cell adhesion molecule (EpCAM) expression and cells are stained with the nucleic acid dye 4′6-diamidino-2-phenylindole (DAPI), Cytokeratin-PE (CK), and CD45-APC. Only DAPI+/CK+ objects are presented to the operator to identify circulating tumor cells (CTC) and the identity of all other cells and potential undetected CTC remains unrevealed. Here, we used the open source imaging program Automatic CTC Classification, Enumeration and PhenoTyping (ACCEPT) to analyze all DAPI+ nuclei in EpCAM-enriched blood samples obtained from 192 metastatic non-small cell lung cancer (NSCLC) patients and 162 controls. Significantly larger numbers of nuclei were detected in 300 patient samples with an average and standard deviation of 73,570 ± 74,948, as compared to 359 control samples with an average and standard deviation of 4191 ± 4463 (p < 0.001). In patients, only 18% ± 21% and in controls 23% ± 15% of the nuclei were identified as leukocytes or CTC. Adding CD16-PerCP for granulocyte staining, the use of an LED as the light source for CD45-APC excitation and plasma membrane staining obtained with wheat germ agglutinin significantly improved the classification of EpCAM-enriched cells, resulting in the identification of 94% ± 5% of the cells. However, especially in patients, the origin of the unidentified cells remains unknown. Further studies are needed to determine if undetected EpCAM+/DAPI+/CK-/CD45- CTC is present among these cells.
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Affiliation(s)
- Sanne de Wit
- Department of Medical Cell BioPhysics, University of Twente, 7522 NH Enschede, The Netherlands.
| | - Leonie L Zeune
- Department of Medical Cell BioPhysics, University of Twente, 7522 NH Enschede, The Netherlands.
- Department of Applied Mathematics, University of Twente, 7522 NH Enschede, The Netherlands.
| | - T Jeroen N Hiltermann
- Department of Pulmonary Diseases, University Medical Center Groningen, University of Groningen, 9713 GZ Groningen, The Netherlands.
| | - Harry J M Groen
- Department of Pulmonary Diseases, University Medical Center Groningen, University of Groningen, 9713 GZ Groningen, The Netherlands.
| | - Guus van Dalum
- Department of General, Visceral and Pediatric Surgery, University Hospital of the Heinrich-Heine-University Düsseldorf, 40225 Düsseldorf, Germany.
| | - Leon W M M Terstappen
- Department of Medical Cell BioPhysics, University of Twente, 7522 NH Enschede, The Netherlands.
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29
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Vermesh O, Aalipour A, Ge TJ, Saenz Y, Guo Y, Alam IS, Park SM, Adelson CN, Mitsutake Y, Vilches-Moure J, Godoy E, Bachmann MH, Ooi CC, Lyons JK, Mueller K, Arami H, Green A, Solomon EI, Wang SX, Gambhir SS. An intravascular magnetic wire for the high-throughput retrieval of circulating tumour cells in vivo. Nat Biomed Eng 2018; 2:696-705. [PMID: 30505627 PMCID: PMC6261517 DOI: 10.1038/s41551-018-0257-3] [Citation(s) in RCA: 78] [Impact Index Per Article: 11.1] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/13/2017] [Accepted: 06/04/2018] [Indexed: 12/31/2022]
Abstract
The detection and analysis of rare blood biomarkers is necessary for early diagnosis of cancer and to facilitate the development of tailored therapies. However, current methods for the isolation of circulating tumour cells (CTCs) or nucleic acids present in a standard clinical sample of only 5-10 ml of blood provide inadequate yields for early cancer detection and comprehensive molecular profiling. Here, we report the development of a flexible magnetic wire that can retrieve rare biomarkers from the subject's blood in vivo at a much higher yield. The wire is inserted and removed through a standard intravenous catheter and captures biomarkers that have been previously labelled with injected magnetic particles. In a proof-of-concept experiment in a live porcine model, we demonstrate the in vivo labelling and single-pass capture of viable model CTCs in less than 10 s. The wire achieves capture efficiencies that correspond to enrichments of 10-80 times the amount of CTCs in a 5-ml blood draw, and 500-5,000 times the enrichments achieved using the commercially available Gilupi CellCollector.
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Affiliation(s)
- Ophir Vermesh
- Molecular Imaging Program at Stanford, Stanford University, Stanford, CA, USA
- Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA
| | - Amin Aalipour
- Molecular Imaging Program at Stanford, Stanford University, Stanford, CA, USA
- Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA
- Department of Bioengineering, Stanford University, Stanford, CA, USA
| | - T Jessie Ge
- Molecular Imaging Program at Stanford, Stanford University, Stanford, CA, USA
- Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA
- Howard Hughes Medical Institute, Chevy Chase, MD, USA
| | - Yamil Saenz
- Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA
| | - Yue Guo
- Department of Electrical Engineering, Stanford University, Stanford, CA, USA
| | - Israt S Alam
- Molecular Imaging Program at Stanford, Stanford University, Stanford, CA, USA
- Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA
| | - Seung-Min Park
- Molecular Imaging Program at Stanford, Stanford University, Stanford, CA, USA
- Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA
| | | | - Yoshiaki Mitsutake
- Division of Cardiovascular Medicine, Stanford University, Stanford, CA, USA
| | - Jose Vilches-Moure
- Department of Comparative Medicine, Stanford University, Stanford, CA, USA
| | - Elias Godoy
- Department of Comparative Medicine, Stanford University, Stanford, CA, USA
| | - Michael H Bachmann
- Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA
- Department of Pediatrics, Stanford University, Stanford, CA, USA
| | - Chin Chun Ooi
- Department of Chemical Engineering, Stanford University, Stanford, CA, USA
| | | | - Kerstin Mueller
- Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA
| | - Hamed Arami
- Molecular Imaging Program at Stanford, Stanford University, Stanford, CA, USA
- Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA
| | - Alfredo Green
- Division of Cardiovascular Medicine, Stanford University, Stanford, CA, USA
| | | | - Shan X Wang
- Department of Electrical Engineering, Stanford University, Stanford, CA, USA
- Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA
| | - Sanjiv S Gambhir
- Molecular Imaging Program at Stanford, Stanford University, Stanford, CA, USA.
- Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA.
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30
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Wu CP, Wu P, Zhao HF, Liu WL, Li WP. Clinical Applications of and Challenges in Single-Cell Analysis of Circulating Tumor Cells. DNA Cell Biol 2018; 37:78-89. [PMID: 29265876 DOI: 10.1089/dna.2017.3981] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Affiliation(s)
- Chang-peng Wu
- Department of Neurosurgery, Shenzhen Second People's Hospital, Clinical Medicine College of Anhui Medical University, Shenzhen, China
- Department of Neurosurgery, Shenzhen Key Laboratory of Neurosurgery, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, Shenzhen, China
| | - Peng Wu
- The Affiliated Luohu Hospital of Shenzhen University, Shenzhen Luohu Hospital Group Department of Urology, Shenzhen, China
| | - Hua-fu Zhao
- Department of Neurosurgery, Shenzhen Key Laboratory of Neurosurgery, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, Shenzhen, China
- Department of Neurosurgery/Neuro-oncology, State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China
| | - Wen-lan Liu
- Department of Neurosurgery, Shenzhen Key Laboratory of Neurosurgery, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, Shenzhen, China
| | - Wei-ping Li
- Department of Neurosurgery, Shenzhen Key Laboratory of Neurosurgery, The First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, Shenzhen, China
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31
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Abraham J, Singh S, Joshi S. Liquid biopsy - emergence of a new era in personalized cancer care. ACTA ACUST UNITED AC 2018. [DOI: 10.1186/s41241-018-0053-0] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022]
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32
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Kuai JH, Wang Q, Zhang AJ, Zhang JY, Chen ZF, Wu KK, Hu XZ. Epidermal growth factor receptor-targeted immune magnetic liposomes capture circulating colorectal tumor cells efficiently. World J Gastroenterol 2018; 24:351-359. [PMID: 29391757 PMCID: PMC5776396 DOI: 10.3748/wjg.v24.i3.351] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/08/2017] [Revised: 11/27/2017] [Accepted: 12/04/2017] [Indexed: 02/06/2023] Open
Abstract
AIM To compare the capacity of newly developed epidermal growth factor receptor (EGFR)-targeted immune magnetic liposomes (EILs) vs epithelial cell adhesion molecule (EpCAM) immunomagnetic beads to capture colorectal circulating tumor cells (CTCs).
METHODS EILs were prepared using a two-step method, and the magnetic and surface characteristics were confirmed. The efficiency of capturing colorectal CTCs as well as the specificity were compared between EILs and EpCAM magnetic beads.
RESULTS The obtained EILs had a lipid nanoparticle structure similar to cell membrane. Improved binding with cancer cells was seen in EILs compared with the method of coupling nano/microspheres with antibody. The binding increased as the contact time extended. Compared with EpCAM immunomagnetic beads, EILs captured more CTCs in peripheral blood from colorectal cancer patients. The captured cells showed consistency with clinical diagnosis and pathology. Mutation analysis showed same results between captured CTCs and cancer tissues.
CONCLUSION EGFR antibody-coated magnetic liposomes show high efficiency and specificity in capturing colorectal CTCs.
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Affiliation(s)
- Jing-Hua Kuai
- Department of Gastroenterology, Qilu Hospital of Shandong University, Qingdao 266035, Shandong Province, China
| | - Qing Wang
- Department of Gastroenterology, Qilu Hospital of Shandong University, Qingdao 266035, Shandong Province, China
| | - Ai-Jun Zhang
- Department of Gastroenterology, Qilu Hospital of Shandong University, Qingdao 266035, Shandong Province, China
| | - Jing-Yu Zhang
- Department of Gastroenterology, Qilu Hospital of Shandong University, Qingdao 266035, Shandong Province, China
| | - Zheng-Feng Chen
- Department of Gastroenterology, Qilu Hospital of Shandong University, Qingdao 266035, Shandong Province, China
| | - Kang-Kang Wu
- Department of Gastroenterology, Qilu Hospital of Shandong University, Qingdao 266035, Shandong Province, China
| | - Xiao-Zhen Hu
- Department of General Surgery, Qilu Hospital of Shandong University, Qingdao 266035, Shandong Province, China
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SUN ZH, ZHOU LH, DENG GJ, ZHEGN MB, YAN WQ, LI WJ, CAI LT, GONG P. Tumor Targeting of Fluorescent Magnetic IR780-Fe 3 O 4 Nanoparticles with for Detection of Circulating Tumor Cells. CHINESE JOURNAL OF ANALYTICAL CHEMISTRY 2017. [DOI: 10.1016/s1872-2040(17)61041-7] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
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Tang Y, Qiao G, Xu E, Xuan Y, Liao M, Yin G. Biomarkers for early diagnosis, prognosis, prediction, and recurrence monitoring of non-small cell lung cancer. Onco Targets Ther 2017; 10:4527-4534. [PMID: 28979144 PMCID: PMC5602468 DOI: 10.2147/ott.s142149] [Citation(s) in RCA: 49] [Impact Index Per Article: 6.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022] Open
Abstract
Despite advances in the management of non-small cell lung cancer, it remains to be the leading cause of cancer-related deaths worldwide primarily because of diagnosis at a late stage with an overall 5-year survival rate of 17%. A reduction in mortality was achieved by low-dose computed tomography screening of high-risk patients. However, the benefit was later challenged by the high false positive rate, resulting in unnecessary follow-ups, thus entailing a burden on both the health care system and the individual. The diagnostic dilemma imposed by imaging modalities has created a need for the development of biomarkers capable of differentiating benign nodules from malignant ones. In the past decade, with the advancements in high-throughput profiling technologies, a huge amount of work has been done to derive biomarkers to supplement clinical diagnosis. However, only a few of them have efficient sensitivity and specificity to be utilized in clinical settings. Therefore, there is an urgent need for the development of sensitive and specific means to detect and diagnose lung cancers at an early stage, when curative interventions are still possible. Due to the invasiveness of tissue biopsies and inability to capture tumor heterogeneity, nowadays enormous efforts have been invested in the development of technologies and biomarkers that enable sensitive and cost-effective testing using substrates that can be obtained in a noninvasive manner. This review, primarily focusing on liquid biopsy, summarizes all documented potential biomarkers for diagnosis, monitoring recurrence treatment response.
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Affiliation(s)
- Yong Tang
- Southern Medical University, Guangzhou, Guangdong Province, China.,Department of Thoracic Surgery, General Hospital of Guangzhou Military Command of PLA, Yuexiu District, Guangzhou City, Guangdong Province, China
| | - Guibin Qiao
- Southern Medical University, Guangzhou, Guangdong Province, China.,Department of Thoracic Surgery, General Hospital of Guangzhou Military Command of PLA, Yuexiu District, Guangzhou City, Guangdong Province, China
| | - Enwu Xu
- Department of Thoracic Surgery, General Hospital of Guangzhou Military Command of PLA, Yuexiu District, Guangzhou City, Guangdong Province, China
| | - Yiwen Xuan
- Department of Thoracic Surgery, General Hospital of Guangzhou Military Command of PLA, Yuexiu District, Guangzhou City, Guangdong Province, China
| | - Ming Liao
- Department of Thoracic Surgery, General Hospital of Guangzhou Military Command of PLA, Yuexiu District, Guangzhou City, Guangdong Province, China
| | - Guilin Yin
- Southern Medical University, Guangzhou, Guangdong Province, China
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Zhou J, Huang A, Yang XR. Liquid Biopsy and its Potential for Management of Hepatocellular Carcinoma. J Gastrointest Cancer 2017; 47:157-67. [PMID: 26969471 DOI: 10.1007/s12029-016-9801-0] [Citation(s) in RCA: 25] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Abstract
PURPOSE We summarized the recent findings of liquid biopsy in cancer field and discussed its potential utility in hepatocellular carcinoma. METHODS Literature published in MEDLINE, EMBASE, and Science Direct electronic databases was searched and reviewed. RESULTS Liquid biopsy specially referred to the detection of nucleic acids (circulating cell-free DNA, cfDNA) and circulating tumor cells (CTCs) in the blood of cancer patients. Compared to conventional single-site sampling or biopsy method, liquid biopsy had the advantages such as non-invasiveness, dynamic monitoring, and the most important of all, overcoming the limit of spatial and temporal heterogeneity. The genomic information of cancer could be profiled by genotyping cfDNA/CTC and subsequently applied to make molecular classification, targeted therapy guidance, and unveil drug resistance mechanisms. The serial sampling feature of liquid biopsy made it possible to monitor treatment response in a real-time manner and predict tumor metastasis/recurrence in advance. CONCLUSIONS Liquid biopsy is a non-invasive, dynamic, and informative sampling method with important clinical translational significance in cancer research and practice. Much work needs to be done before it is used in the management of HCC.
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Affiliation(s)
- Jian Zhou
- Liver Surgery Department, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, 20032, China.
| | - Ao Huang
- Liver Surgery Department, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, 20032, China
| | - Xin-Rong Yang
- Liver Surgery Department, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, 20032, China
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Swennenhuis JF, van Dalum G, Zeune LL, Terstappen LWMM. Improving the CellSearch® system. Expert Rev Mol Diagn 2016; 16:1291-1305. [PMID: 27797592 DOI: 10.1080/14737159.2016.1255144] [Citation(s) in RCA: 48] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
INTRODUCTION The CellSearch® CTC test enumerates tumor cells present in 7.5 ml blood of cancer patients. improvements, extensions and different utilities of the cellsearch system are discussed in this paper. Areas covered: This paper describes work performed with the CellSearch system, which go beyond the normal scope of the test. All results from searches with the search term 'CellSearch' from Web of Science and PubMed were categorized and discussed. Expert commentary: The CellSearch Circulating Tumor Cell test captures and identifies tumor cells in blood that are associated with poor clinical outcome. How to best use CTC in clinical practice is being explored in many clinical trials. The ability to extract information from the CTC to guide therapy will expand the potential clinical utility of CTC.
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Affiliation(s)
- J F Swennenhuis
- a Medical Cell BioPhysics , University of Twente , Enschede , The Netherlands
| | - G van Dalum
- a Medical Cell BioPhysics , University of Twente , Enschede , The Netherlands
| | - L L Zeune
- a Medical Cell BioPhysics , University of Twente , Enschede , The Netherlands
| | - L W M M Terstappen
- a Medical Cell BioPhysics , University of Twente , Enschede , The Netherlands
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Hanssen A, Wagner J, Gorges TM, Taenzer A, Uzunoglu FG, Driemel C, Stoecklein NH, Knoefel WT, Angenendt S, Hauch S, Atanackovic D, Loges S, Riethdorf S, Pantel K, Wikman H. Characterization of different CTC subpopulations in non-small cell lung cancer. Sci Rep 2016; 6:28010. [PMID: 27302574 PMCID: PMC4908396 DOI: 10.1038/srep28010] [Citation(s) in RCA: 72] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/26/2016] [Accepted: 05/27/2016] [Indexed: 01/18/2023] Open
Abstract
Circulating tumour cells (CTCs) serve as valuable biomarkers. However, EpCAM positive CTCs are less frequently detected in NSCLC patients compared to other epithelial tumours. First, EpCAM protein expression was analysed in primary and metastatic lung cancer tissue. In both groups 21% of the samples were EpCAM negative. Second, the CellSearch system identified 15% of patients (n = 48) as CTC positive whereas a multiplex RT-PCR for PIK3CA, AKT2, TWIST, and ALDH1 following EGFR, HER2 and EpCAM based enrichment detected CTCs in 29% of the patients. Interestingly, 86% of CTC positive patients were found to express ALDH1. Only 11% of the patients were CTC-positive by both techniques. CTC positivity was associated with patient disease state when assessed by the multiplex RT-PCR assay (p = 0.015). Patients harbouring tumours with an altered EGFR genotype were more frequently CTC-positive compared to patients with EGFR wildtype tumours. In subsets of patients, CTCs were found to express genes involved in resistance to therapy such as HER3 and MET. In conclusion, using multiple targets for CTC capture and identification increases the sensitivity of CTC detection in NSCLC patients, which can be explained by the presence of different CTC subtypes with distinct molecular features.
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Affiliation(s)
- Annkathrin Hanssen
- Department of Tumour Biology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
| | | | - Tobias M Gorges
- Department of Tumour Biology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
| | - Aline Taenzer
- Department of Tumour Biology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany.,Center for Neurology, Neurosurgery, and Psychiatry, Department of Psychiatry, Campus Benjamin Franklin, Charité University Hospital Berlin, Germany
| | - Faik G Uzunoglu
- Department of General, Visceral and Thoracic Surgery, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
| | - Christiane Driemel
- Department of General, Visceral and Pediatric Surgery, University Hospital and Medical Faculty of the Heinrich-Heine University Düsseldorf, Düsseldorf, Germany
| | - Nikolas H Stoecklein
- Department of General, Visceral and Pediatric Surgery, University Hospital and Medical Faculty of the Heinrich-Heine University Düsseldorf, Düsseldorf, Germany
| | - Wolfram T Knoefel
- Department of General, Visceral and Pediatric Surgery, University Hospital and Medical Faculty of the Heinrich-Heine University Düsseldorf, Düsseldorf, Germany
| | - Sebastian Angenendt
- Department of General, Visceral and Pediatric Surgery, University Hospital and Medical Faculty of the Heinrich-Heine University Düsseldorf, Düsseldorf, Germany
| | | | - Djordje Atanackovic
- Department of Internal Medicine II and Clinic (Oncology Centre), University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
| | - Sonja Loges
- Department of Tumour Biology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany.,Department of Internal Medicine II and Clinic (Oncology Centre), University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
| | - Sabine Riethdorf
- Department of Tumour Biology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
| | - Klaus Pantel
- Department of Tumour Biology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
| | - Harriet Wikman
- Department of Tumour Biology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
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Lianidou ES. Gene expression profiling and DNA methylation analyses of CTCs. Mol Oncol 2016; 10:431-42. [PMID: 26880168 DOI: 10.1016/j.molonc.2016.01.011] [Citation(s) in RCA: 39] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/10/2015] [Revised: 01/19/2016] [Accepted: 01/25/2016] [Indexed: 01/26/2023] Open
Abstract
A variety of molecular assays have been developed for CTCs detection and molecular characterization. Molecular assays are based on the nucleic acid analysis in CTCs and are based on total RNA isolation and subsequent mRNA quantification of specific genes, or isolation of genomic DNA that can be for DNA methylation studies and mutation analysis. This review is mainly focused on gene expression and methylation studies in CTCs in various types of cancer.
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Affiliation(s)
- Evi S Lianidou
- Analysis of Circulating Tumor Cells Lab, Lab of Analytical Chemistry, Department of Chemistry, University of Athens, 15771, Greece.
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Schlange T, Pantel K. Potential of circulating tumor cells as blood-based biomarkers in cancer liquid biopsy. Pharmacogenomics 2016; 17:183-6. [PMID: 26799583 DOI: 10.2217/pgs.15.163] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/28/2022] Open
Affiliation(s)
- Thomas Schlange
- Bayer Pharma AG, Global Biomarker Research, 42096 Wuppertal, Germany
| | - Klaus Pantel
- Department of Tumour Biology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany
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40
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Andree KC, van Dalum G, Terstappen LWMM. Challenges in circulating tumor cell detection by the CellSearch system. Mol Oncol 2015; 10:395-407. [PMID: 26795350 DOI: 10.1016/j.molonc.2015.12.002] [Citation(s) in RCA: 225] [Impact Index Per Article: 22.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/16/2015] [Revised: 11/19/2015] [Accepted: 12/03/2015] [Indexed: 12/11/2022] Open
Abstract
Enumeration and characterization of circulating tumor cells (CTC) hold the promise of a real time liquid biopsy. They are however present in a large background of hematopoietic cells making their isolation technically challenging. In 2004, the CellSearch system was introduced as the first and only FDA cleared method designed for the enumeration of circulating tumor cells in 7.5 mL of blood. Presence of CTC detected by CellSearch is associated with poor prognosis in metastatic carcinomas. CTC remaining in patients after the first cycles of therapy indicates a futile therapy. Here we review challenges faced during the development of the CellSearch system and the difficulties in assigning objects as CTC. The large heterogeneity of CTC and the different approaches introduced in recent years to isolate, enumerate and characterize CTC results in a large variation of the number of CTC reported urging the need for uniform definitions and at least a clear definition of what the criteria are for assigning an object as a CTC.
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Affiliation(s)
- Kiki C Andree
- Department of Medical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The Netherlands
| | - Guus van Dalum
- Department of Medical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The Netherlands
| | - Leon W M M Terstappen
- Department of Medical Cell BioPhysics, University of Twente, Hallenweg 23, 7522 NH Enschede, The Netherlands.
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