Simultaneous biochemical and functional phenotyping of single circulating tumor cells using ultrahigh throughput and recovery microfluidic devices

Lab on a Chip ◽  
2021 ◽  
Author(s):  
Yang Liu ◽  
Wujun Zhao ◽  
Rui Cheng ◽  
Jamie Hodgson ◽  
Mary Egan ◽  
...  
Keyword(s):  
Cancer Patients ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Microfluidic Devices ◽  
Tumour Cells ◽  
Circulating Tumour Cells ◽  
Dynamic Nature ◽  
Blood Samples

Profiling circulating tumour cells (CTCs) in cancer patients’ blood samples is critical to understanding the complex and dynamic nature of metastasis. This task is challenged by the fact that CTCs...

Molecular markers for circulating tumor cells in breast cancer.

2011 ◽  
Vol 29 (27_suppl) ◽  
pp. 223-223
Author(s):  
R. Zeillinger ◽  
E. Obermayr ◽  
A. Fink-Retter ◽  
G. Heinze ◽  
A. Reinthaller ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Gene Panel ◽  
Control Group ◽  
Gene Marker ◽  
Breast Cancer Patients ◽  
Gene Markers ◽  
Blood Samples

223 Background: Recently, we identified a six gene panel (CCNE2, DKFZp762E1312, EMP2, MAL2, PPIC, and SLC6A8) for the RT-qPCR based detection of circulating tumor cells (CTC) in breast cancer patients. The aim of the present study was to evaluate the gene panel in further blood samples. Methods: Blood samples were taken from breast cancer patients with metastatic disease (MBC, N=10) or with no evidence of disease (NED, N=30). Putative CTC were enriched by Oncoquick density gradient centrifugation. Total RNA was isolated with RNeasy Micro Kit (QIAgen). Template cDNA was generated with M-MLV Reverse Transcriptase, RNase H Minus (Promega) and random nonamers as primers. RT-qPCR was performed in duplicate reactions using TaqMan Assays (Applied Biosystems) with default thermal cycling parameters. Raw data were analyzed with the AB7900 Sequence Detection Software version 2.2.2 using automatic baseline correction and manual cycle threshold setting. Gene expression was normalized to GAPDH expression. A threshold value TX for each gene X was set at two standard deviations above the mean dCtX value in the healthy control group. A patient was defined as CTC-positive, if at least one gene marker was over-expressed compared to the defined threshold. Results: The gene panel consisting of CCNE2, DKFZp762E1312, EMP2, MAL2, PPIC, and SLC6A8 identified 4/11 MBC but only 5/27 NED patients as CTC positive (p=0.163). By adding known CTC markers (SCGB2A2, TFF1, FXYD3, AGR2, S100A18, and EPCAM) to the panel, 7/11 MBC but only 6/27 NED patients were CTC positive (p=0.018). The presence of CTC in NED patients correlated with pN staging (p=0.026). Only one out of the six CTC positive NED patients relapsed within the observation period (median 35 months, range 25-39 months from blood sampling). We observed no correlation of CTC positivity and recurrence in NED patients. Conclusions: The sensitivity of the RT-qPCR based CTC detection in breast cancer patients may be enhanced by adding known CTC markers (SCGB2A2, TFF1, FXYD3, AGR2, S100A18, and EPCAM) to the six gene panel (CCNE2, DKFZp762E1312, EMP2, MAL2, PPIC, and SLC6A8). Longer follow-up times are needed to evaluate the predictive value of the gene markers on survival.

scholarly journals Quantitation of circulating tumor cells in blood samples from ovarian and prostate cancer patients using tumor-specific fluorescent ligands

2008 ◽  
Vol 123 (8) ◽  
pp. 1968-1973 ◽  
Author(s):  
Wei He ◽  
Sumith A. Kularatne ◽  
Kimberly R. Kalli ◽  
Franklyn G. Prendergast ◽  
Robert J. Amato ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Blood Samples ◽  
Fluorescent Ligands

scholarly journals Isolation of Circulating Tumor Cells from Peripheral Blood Samples of Cancer Patients Using Microfluidic Technology

2020 ◽  
Vol 12 (6) ◽  
pp. 62
Author(s):  
A.B. Volovetskiy ◽  
P.A. Malinina ◽  
A.Y. Kapitannikova ◽  
S.V. Smetanina ◽  
I.A. Kruglova ◽  
...  
Keyword(s):  
Cancer Patients ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Peripheral Blood ◽  
Blood Samples ◽  
Microfluidic Technology

Positively Charged Magnetic Nanoparticles for Capture of Circulating Tumor Cells from Clinical Blood Samples

Nano LIFE ◽  
2020 ◽  
Vol 10 (03) ◽  
pp. 1971001 ◽  
Author(s):  
Shengming Wu ◽  
Yilong Wang ◽  
Donglu Shi
Keyword(s):  
Magnetic Nanoparticles ◽  
Cancer Cells ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cancer Cell ◽  
Detection Sensitivity ◽  
Blood Samples ◽  
Charged Nanoparticles ◽  
Positively Charged

Isolation of circulating tumor cells (CTCs) from cancer patients is of high value for disease monitoring and metastasis diagnosis. Although many new detection methods have emerged in recent years, the detection of CTCs is a current challenge due to lack of specific and sensitive markers. In our previous work, cancer cell surfaces, from over 20 cancer cell lines, have been shown to be negatively-charged regardless of their phenotype by using electrically-charged nanoparticles as a probe. The strong electrostatic interaction between the negative cancer cells and positively charged nanoparticles can well remain in a physiological liquid environment in the presence of serum proteins, enabling effective binding between them. As a result, the cancer cells can be magnetically separated by employing an external magnet. In this technical report, we present preliminary results on the investigation of CTC isolation from both mimetic and clinical blood samples. We show high CTC detection sensitivity by the positively-charged magnetic nanoparticles (PMNs) even at the original concentration of 10 cells per mL mimetic blood sample. The CTCs in the peripheral blood of colorectal cancer patients were isolated and identified by cellular morphology and immunofluorescence staining.

scholarly journals Distribution and Clinical Analysis of EpCAM+/Vimentin+ Circulating Tumor Cells in High-Risk Population and Cancer Patients

2021 ◽  
Vol 11 ◽  
Author(s):  
Chunjin Huang ◽  
Sheng Ding ◽  
Chunyan Huang ◽  
Feng Pan ◽  
Xiaodong Liu ◽  
...  
Keyword(s):  
High Risk ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Peripheral Blood ◽  
Healthy Subjects ◽  
Magnetic Bead ◽  
High Risk Population ◽  
Risk Population ◽  
Blood Samples

Circulating Tumor Cells (CTCs) are already present in the peripheral blood of patients with early tumors and even precancerous lesions. The objective of this study was to determine the count of CTCs in peripheral blood from high-risk population(HRP), healthy subjects and patients with Pan-cancer. The CTCs in the peripheral blood from HRP and cancer patients were enriched and identified based on the positive sorting method by epithelial cell adhesion molecular (EpCAM) liposome magnetic bead (Ep-LMB) and Vimentin liposome magnetic bead (Vi-LMB). Simultaneously, further analysis was carried out focusing on the clinical characteristics of patients by collecting the peripheral blood samples from healthy subjects as the parallel control. According to the results, the prepared LMBs had high specificity and stability, resulting in an average (Av) proliferation rate of over 90% for each cell line, and the average capture rate of higher than 80%. In terms of CTCs count detection in clinical blood samples, the average count was 0.9 (Ep: Av=0.6, Vi: Av=0.3), 2.4 (Ep: Av=1.4, Vi: Av=0.8) and 7.3 (Ep: Av=4.0, Vi: Av=3.3) in healthy subjects, HRP and total cancer patients, respectively. Besides, there was no obvious difference in the average count of CTCs among patients with different cancer types. While count of CTCs in the aforementioned cancer patients was statistically different from that in healthy subjects and patients with HRP. The survival time of cancer patients whose number of CTCs is greater than the average is significantly increased. Collectively, the study confirmed that CTCs can achieve early tumor detection and auxiliary diagnosis, and its number is related to the occurrence and development of tumors, and CTCs can be detected in HRP and sub-health population.

PS02.071: EVALUATION OF CIRCULATING TUMOR CELLS IN ESOPHAGEAL CANCER PATIENTS

2018 ◽  
Vol 31 (Supplement_1) ◽  
pp. 140-140
Author(s):  
Daisuke Ujiie ◽  
Shinji Ohki ◽  
Hirokazu Okayama ◽  
Leo Yamada ◽  
Takeshi Tada ◽  
...  
Keyword(s):  
Esophageal Cancer ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Treatment Effect ◽  
Conflicts Of Interest ◽  
Malignant Tumors ◽  
Metastatic Breast ◽  
Blood Samples ◽  
Before And After

Abstract Background Circulating tumor cells (CTCs) in patients with malignant tumors can be used as a prognostic marker. Recently the existence of mesenchymal CTCs have been detected by new methods. However, it has been not yet clarified how CTCs are associated with the treatment effects in esophageal cancer patients. We assessed CTCs in esophageal cancer patients and investigated the relationship between CTCs and treatment effect. Methods Seven patients who had potential of curative resection have been enrolled and peripheral blood samples (10ml) were collected before and after treatment. All patients received chemotherapy (5-FU and cisplatin) and four patients of them received as neoadjuvant therapy. Other patients received only chemotherapy and radiation therapy without operation. CTCs were analyzed using a Microfluidic Chip devise provide with the Nihon Gene Research Laboratories. This system can isolates CTCs from blood samples, based on their size and deformability differences from blood cells. Phenotypes of CTCs are determined by staining and scanning systems. A previous report revealed that this method had higher sensitivity for CTCs than conventional methods (the Cell Search system) in 61 metastatic breast cancer. Results Treatments effects were stable disease or better in all cases. Multiple CTCs were detected in all cases before treatment. Five patients had epithelial CTCs and others had only mesenchymal CTCs. Total number of CTCs after treatment whose data can be available decreased except for two cases. One patient had no CTC after treatment. Two cases have been currently analyzing. Conclusion CTCs may exist almost all patients of Stage II or more esophageal cancer. Decreasing the number of CTCs after treatment suggests some relationship between CTCs and treatment effect, and the accumulation of more cases is necessary. Disclosure All authors have declared no conflicts of interest.

Optical cytosensors for detection of circulating tumour cells

2022 ◽  
Author(s):  
Fereshteh Vajhadin ◽  
Mohammad Mazloum ◽  
Alireza Sanati ◽  
Reyhaneh Haghniaz ◽  
Jadranka Travas-Sejdic
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Tumor Markers ◽  
Heart Defects ◽  
Tumour Cells ◽  
Blood Analysis ◽  
Circulating Tumour Cells

Blood analysis is an established approach to monitor various diseases, ranging from heart defects and diabetes to cancer. Among various tumor markers in the blood, circulating tumor cells (CTCs) have...

Υγρή βιοψία

2019 ◽  
Author(s):  
Μάρθα Ζαβρίδου
Keyword(s):  
Nucleic Acids ◽  
Cancer Treatment ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Circulating Tumor Dna ◽  
Tumour Cells ◽  
Full Length ◽  
Circulating Tumour Cells ◽  
Roche Diagnostics ◽  
Tumor Dna

Η «υγρή βιοψία», βασισμένη στην ανάλυση των κυκλοφορούντων καρκινικών κυττάρων (circulating tumor cells, CTCs), του εξωκυττάριου καρκινικού DNA (circulating tumor DNA, ctDNA), των miRNAs και των εξωσωμάτων παρέχει μια μη-επεμβατική παρακολούθηση της εξέλιξης της νόσου και της αποτελεσματικότητας της θεραπείας, σε πραγματικό χρόνο.Στην παρούσα διδακτορική διατριβή, αρχικά αξιολογήσαμε την επίδραση των προαναλυτικών συνθηκών για την ανάλυση γονιδιακής έκφρασης στα CTCs. Για το σκοπό αυτό πραγματοποιήθηκαν πειράματα εμβολιασμού 100 MCF7 σε σωληνάρια συλλογής περιφερικού αίματος, με διαφορετική σύσταση αντιπηκτικού και συντηρητικών των κυττάρων. Στη συνέχεια ακολούθησε η απομόνωση των CTCs με EpCAM-θετικό ανοσομαγνητικό εμπλουτισμό σε διαστήματα 0h, 24h και 48h και ποσοτικός προσδιορισμός της έκφρασης των γονιδίων B2M και CK-19 στα CTCs με RT-qPCR. Παράλληλα, προτείνουμε μια ολοκληρωμένη διαδικασία ελέγχου ποιότητας όλων των βημάτων που συμπεριλαμβάνονται στην πειραματική πορεία για αναλύσεις γονιδιακής έκφρασης σε δείγματα υγρής βιοψίας. Τα αποτελέσματά μας δείχνουν ότι οι αναλύσεις των CTCs σε επίπεδο mRNA επηρεάζονται από τα συντηρητικά ή/και τα αντιπηκτικά που χρησιμοποιούνται στα περισσότερα σωληνάρια συλλογής περιφερικού αίματος.Στη συνέχεια, στόχο μας αποτέλεσε η επιλογή του βέλτιστου συστήματος απομόνωσης/εμπλουτισμού CTCs για το μοριακό χαρακτηρισμό τους στον HNSCC. Για αυτό το λόγο, πραγματοποιήθηκε για πρώτη φορά άμεση σύγκριση του συστήματος Parsortix, που βασίζεται σε σύστημα μικροροών για την απομόνωση/εμπλουτισμό των CTCs με βάση το μέγεθός τους, με τον ανοσομαγνητικό εμπλουτισμό των CTCs με βάση το EpCAM από το ίδιο περιφερικό αίμα ασθενών με HNSCC και προχωρήσαμε στη συνέχεια σε μοριακό χαρακτηρισμό των CTCs σε επίπεδο γονιδιακής έκφρασης. Τα αποτελέσματα της παρούσας μελέτης υποδεικνύουν σαφώς ότι ο πληθυσμός των CTCs που απομονώθηκε με το σύστημα Parsortix είναι υψηλότερης καθαρότητας σε σχέση με τον πληθυσμό των CTCs που απομονώθηκε με EpCAM θετικό ανοσομαγνητικό εμπλουτισμό. Επιπρόσθετα, αναπτύξαμε μεθοδολογία πολλαπλής RT-qPCR για τον ταυτόχρονο ποσοτικό προσδιορισμό της έκφρασης του υποδοχέα των ανδρογόνων (AR), AR-full length (AR-FL), των εναλλακτικών μεταγράφων του AR-V7 και AR-567 και του ολικού AR-total με ανιχνευτές υδρόλυσης στο όργανο Cobas z480 (Roche Diagnostics). Η αναπτυχθείσα μεθοδολογία εφαρμόστηκε και αξιολογήθηκε κλινικά στα CTCs και στα εξωσώματα που απομονώθηκαν από το πλάσμα ασθενών με μεταστατικό ευνουχοάντοχο καρκίνο προστάτη. Τα αποτελέσματά μας υποδεικνύουν ξεκάθαρα ότι η μέθοδος μπορεί να εφαρμοστεί τόσο στα CTCs όσο και στα εξωσώματα.Στο τελευταίο μέρος της διατριβής, διερευνήθηκε το μοριακό προφίλ των CTCs και των εξωσωμάτων ασθενών με ευνουχοάντοχο καρκίνο προστάτη. Πιο συγκεκριμένα, μελετήθηκε το προφίλ της έκφρασης των γονιδίων PD-L1, CK-19, των εναλλακτικών μεταγράφων AR-FL, AR-V7, AR-567 και AR-total, καθώς και το προφίλ μεθυλίωσης των γονιδίων GSTP1 και RASSF1A. Η διδακτορική διατριβή χρηματοδοτήθηκε από το Ευρωπαϊκό Πρόγραμμα IMI-CANCER-ID (αρ. συμβολαίου 115749): “Cancer treatment and monitoring through identification of circulating tumour cells and tumour related nucleic acids in blood” (https://www.cancer-id.eu/)

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