scholarly journals Optimization of rVAR2-Based Isolation of Cancer Cells in Blood for Building a Robust Assay for Clinical Detection of Circulating Tumor Cells

2020 ◽  
Vol 21 (7) ◽  
pp. 2401 ◽  
Author(s):  
Nicolai T. Sand ◽  
Tobias B. Petersen ◽  
Sara R. Bang-Christensen ◽  
Theresa D. Ahrens ◽  
Caroline Løppke ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cancer Cell ◽  
Cancer Progression ◽  
Magnetic Beads ◽  
Liquid Biopsies ◽  
Blood Samples ◽  
Non Invasive ◽  
Clinical Detection

Early detection and monitoring of cancer progression is key to successful treatment. Therefore, much research is invested in developing technologies, enabling effective and valuable use of non-invasive liquid biopsies. This includes the detection and analysis of circulating tumor cells (CTCs) from blood samples. Recombinant malaria protein VAR2CSA (rVAR2) binds a unique chondroitin sulfate modification present on the vast majority of cancers and thereby holds promise as a near-universal tumor cell-targeting reagent to isolate CTCs from complex blood samples. This study describes a technical approach for optimizing the coupling of rVAR2 to magnetic beads and the development of a CTC isolation platform targeting a range of different cancer cell lines. We investigate both direct and indirect approaches for rVAR2-mediated bead retrieval of cancer cells and conclude that an indirect capture approach is most effective for rVAR2-based cancer cell retrieval.

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 Clinical Applications of Circulating Tumor Cells and Circulating Tumor DNA as a Liquid Biopsy Marker in Colorectal Cancer

Cancers ◽  
2021 ◽  
Vol 13 (18) ◽  
pp. 4500
Author(s):  
Isabel Heidrich ◽  
Thaer S. A. Abdalla ◽  
Matthias Reeh ◽  
Klaus Pantel
Keyword(s):  
Colorectal Cancer ◽  
Disease Progression ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Liquid Biopsy ◽  
Personalized Therapy ◽  
Liquid Biopsies ◽  
Curative Surgery ◽  
Non Invasive ◽  
Staging Systems

Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide. It is a heterogeneous tumor with a wide genomic instability, leading to tumor recurrence, distant metastasis, and therapy resistance. Therefore, adjunct non-invasive tools are urgently needed to help the current classical staging systems for more accurate prognostication and guiding personalized therapy. In recent decades, there has been an increasing interest in the diagnostic, prognostic, and predictive value of circulating cancer-derived material in CRC. Liquid biopsies provide direct non-invasive access to tumor material, which is shed into the circulation; this enables the analysis of circulating tumor cells (CTC) and genomic components such as circulating free DNA (cfDNA), which could provide the key for personalized therapy. Liquid biopsy (LB) allows for the identification of patients with a high risk for disease progression after curative surgery, as well as longitudinal monitoring for disease progression and therapy response. Here, we will review the most recent studies on CRC, demonstrating the clinical potential and utility of CTCs and ctDNA. We will discuss some of the advantages and limitations of LBs and the future perspectives in the field of CRC management.

scholarly journals Activated fibroblasts enhance cancer cell migration by microvesicles-mediated transfer of Galectin-1

2021 ◽  
Author(s):  
Alessandra Toti ◽  
Alice Santi ◽  
Elisa Pardella ◽  
Ilaria Nesi ◽  
Richard Tomasini ◽  
...  
Keyword(s):  
Cell Migration ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Cancer Cell ◽  
Cancer Progression ◽  
Tumor Stroma ◽  
Activation Process ◽  
Cancer Cell Migration ◽  
Stromal Compartment ◽  
Activated Fibroblasts

AbstractCancer-associated fibroblasts (CAFs) are one of the main components of the stromal compartment in the tumor microenvironment (TME) and the crosstalk between CAFs and cancer cells is essential for tumor progression and aggressiveness. Cancer cells mediate an activation process, converting normal fibroblasts into CAFs, that are characterized by modified expression of many proteins and increased production and release of microvesicles (MVs), extracellular vesicles generated by outwards budding from the cell membrane. Recent evidence underlined that the uptake of CAF-derived MVs changes the overall protein content of tumor cells. In this paper, we demonstrate that tumor activated fibroblasts overexpress Galectin-1 (Gal-1) and consequently release MVs containing increased levels of this protein. The uptake of Gal-1 enriched MVs by tumor cells leads to the upregulation of its intracellular concentration, that strongly affects cancer cell migration, while neither proliferation nor adhesion are altered. Accordingly, tumor cells co-cultured with fibroblasts silenced for Gal-1 have a reduced migratory ability. The present work reveals the key role of an exogenous protein, Gal-1, derived from activated fibroblasts, in cancer progression, and contributes to clarify the importance of MVs-mediated protein trafficking in regulating tumor-stroma crosstalk.

scholarly journals Enrichment of Circulating Tumor Cells from Whole Blood Using a Microfluidic Device for Sequential Physical and Magnetophoretic Separations

Micromachines ◽  
2020 ◽  
Vol 11 (5) ◽  
pp. 481 ◽  
Author(s):  
Jusin Lee ◽  
Onejae Sul ◽  
Seung-Beck Lee
Keyword(s):  
Specific Surface ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Microfluidic Device ◽  
Whole Blood ◽  
Blood Cells ◽  
Magnetic Beads ◽  
Separation Efficiency ◽  
White Blood Cells

Based on their high clinical potential, the isolation and enrichment of rare circulating tumor cells (CTCs) from peripheral blood cells has been widely investigated. There have been technical challenges with CTC separation methods using solely cancer-specific surface molecules or just using physical properties of CTCs, as they may suffer from heterogeneity or lack of specificity from overlapping physical characteristics with leukocytes. Here, we integrated an immunomagnetic-based negative enrichment method that utilizes magnetic beads attached to leukocyte-specific surface antigens, with a physical separation method that utilizes the distinct size and deformability of CTCs. By manipulating the pressure distribution throughout the device and balancing the drag and magnetic forces acting on the magnetically labeled white blood cells (WBCs), the sequential physical and magnetophoretic separations were optimized to isolate intact cancer cells, regardless of heterogeneity from whole blood. Using a breast cancer cell line in whole blood, we achieved 100% separation efficiency for cancer cells and an average of 97.2% for WBCs, which resulted in a 93.3% average separation purity. The experimental results demonstrated that our microfluidic device can be a promising candidate for liquid biopsy and can be a vital tool for aiding future cancer research.

scholarly journals CD44v6 Defines a New Population of Circulating Tumor Cells Not Expressing EpCAM

Cancers ◽  
2021 ◽  
Vol 13 (19) ◽  
pp. 4966
Author(s):  
Guillaume Belthier ◽  
Zeinab Homayed ◽  
Fanny Grillet ◽  
Christophe Duperray ◽  
Julie Vendrell ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cell Sorting ◽  
Magnetic Beads ◽  
Therapeutic Response ◽  
Breast Cancer Patients ◽  
Blood Samples ◽  
Different Origins ◽  
Immune Detection

Circulating tumor cells (CTCs) are promising diagnostic and prognostic tools for clinical use. In several cancers, including colorectal and breast, the CTC load has been associated with a therapeutic response as well as progression-free and overall survival. However, counting and isolating CTCs remains sub-optimal because they are currently largely identified by epithelial markers such as EpCAM. New, complementary CTC surface markers are therefore urgently needed. We previously demonstrated that a splice variant of CD44, CD44 variable alternative exon 6 (CD44v6), is highly and specifically expressed by CTC cell lines derived from blood samples in colorectal cancer (CRC) patients. Two different approaches—immune detection coupled with magnetic beads and fluorescence-activated cell sorting—were optimized to purify CTCs from patient blood samples based on high expressions of CD44v6. We revealed the potential of the CD44v6 as a complementary marker to EpCAM to detect and purify CTCs in colorectal cancer blood samples. Furthermore, this marker is not restricted to colorectal cancer since CD44v6 is also expressed on CTCs from breast cancer patients. Overall, these results strongly suggest that CD44v6 could be useful to enumerate and purify CTCs from cancers of different origins, paving the way to more efficacious combined markers that encompass CTC heterogeneity.

scholarly journals Isolation of Circulating Tumor Cells from Glioblastoma Patients by Direct Immunomagnetic Targeting

2020 ◽  
Vol 10 (9) ◽  
pp. 3338 ◽  
Author(s):  
David Lynch ◽  
Branka Powter ◽  
Joseph William Po ◽  
Adam Cooper ◽  
Celine Garrett ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Blood Brain Barrier ◽  
Brain Cancer ◽  
Brain Barrier ◽  
Blood Samples ◽  
Blood Brain

Glioblastoma (GBM) is the most common form of primary brain cancer in adults and tissue biopsies for diagnostic purposes are often inaccessible. The postulated idea that brain cancer cells cannot pass the blood–brain barrier to form circulating tumor cells (CTCs) has recently been overthrown and CTCs have been detected in the blood of GBM patients albeit in low numbers. Given the potential of CTCs to be analyzed for GBM biomarkers that may guide therapy decisions it is important to define methods to better isolate these cells. Here, we determined markers for immunomagnetic targeting and isolation of GBM-CTCs and confirmed their utility for CTC isolation from GBM patient blood samples. Further, we identified a new marker to distinguish isolated GBM-CTCs from residual lymphocytes.

Cancer Cell Surface Negative Charges: A Bio-Physical Manifestation of the Warburg Effect

Nano LIFE ◽  
2017 ◽  
Vol 07 (03n04) ◽  
pp. 1771001 ◽  
Author(s):  
Donglu Shi
Keyword(s):  
Early Detection ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cancer Cell ◽  
Cell Detection ◽  
Cell Capture ◽  
Metabolic Pattern ◽  
Early Cancer Diagnosis ◽  
Highly Sensitive

The early detection of circulating tumor cells (CTCs) in blood as part of medical diagnosis will give the doctors a head start in the provision and treatment of cancer, and therefore, with the advance in Nano technology, there is an increasing expectation of some form of early detection of circulating tumor cells at a highly sensitive level, without any biomarkers, for both early cancer diagnosis and monitoring disease progression after medical intervention. This technical note reports on the recent development in detection of highly sensitive detection of cancer cells without biomarkers. This novel concept is developed based on a hallmark cancer metabolic pattern: high glycolysis rate. Secretion of high level of lactate acid by cancer cells ultimately results in negative electrical charges on their surfaces, enabling strong binding and capturing by the positively-charged nanoprobes, and subsequent magnetic separation. When nanoprobes are incubated with cancer cells in suspension, binding takes place due to charge differences, and cancer cells are then magnetically separated. The separated cells are enumerated using a flow cytometry and identified by pathological and genome sequencing methods. Preliminary results using the approach have shown exceptionally high cancer cell capture rates, therefore potentially applicable in cancer cell detection in clinical settings.

Detection of circulating tumor cells based on improved SERS-active magnetic nanoparticles

2019 ◽  
Vol 11 (22) ◽  
pp. 2918-2928 ◽  
Author(s):  
Ting Xue ◽  
Siqi Wang ◽  
Guoyu Ou ◽  
Yong Li ◽  
Huimin Ruan ◽  
...  
Keyword(s):  
Magnetic Nanoparticles ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Molecular Analysis ◽  
Cell Expansion ◽  
Magnetic Nanoparticle ◽  
Blood Samples ◽  
Phenotype Identification

A SERS-active magnetic nanoparticle was developed for capture, enrichment and detection of cancer cells from blood samples. The isolated CTCs can also be released for further cell expansion, phenotype identification and molecular analysis.

scholarly journals Do malignant cells sleep at night?

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Luis Enrique Cortés-Hernández ◽  
Zahra Eslami-S ◽  
Antoine M. Dujon ◽  
Mathieu Giraudeau ◽  
Beata Ujvari ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cancer Progression ◽  
Liquid Biopsy ◽  
Biological Rhythms ◽  
Clinical Implications ◽  
Malignant Cells ◽  
Metastatic Cascade ◽  
Key Players

AbstractBiological rhythms regulate the biology of most, if not all living creatures, from whole organisms to their constitutive cells, their microbiota, and also parasites. Here, we present the hypothesis that internal and external ecological variations induced by biological cycles also influence or are exploited by cancer cells, especially by circulating tumor cells, the key players in the metastatic cascade. We then discuss the possible clinical implications of the effect of biological cycles on cancer progression, and how they could be exploited to improve and standardize methods used in the liquid biopsy field.

3D morphometric analysis to yield high sensitivity and specificity enumeration of circulating tumor cells.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. e22031-e22031
Author(s):  
Michael Meyer ◽  
Thomas Neumann ◽  
Rahul Katdare ◽  
Chris Presley ◽  
Jon Hayenga ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cancer Cell ◽  
Blood Cells ◽  
Breast Cancer Cells ◽  
Cell Detection ◽  
Normal Cells ◽  
Adaptive Boosting

e22031 Background: The enumeration of circulating tumor cells (CTCs) has become a common method to assess cancer treatment efficacy. In this study, we propose to test our ability to detect and enumerate CTCs using a morphometric classifier based on 3D cell features to distinguish normal cells from lysed blood and breast cancer cells. Methods: Commercially available normal whole blood was lysed to destroy red blood cells, leaving white blood cells and some cell debris. These cells and breast cancer cells from the SK-BR-3 cell line were studied. The cells were stained with hematoxylin, then were automatically imaged using VisionGate’s 3D cell imaging system (Cell-CT) that unambiguously renders individual cells with isometric, sub-micron resolution. Image libraries of cells were created. For each cell, 683 morphology features were computed. Features characterize various aspects of the cell such as chromatin distribution in the nucleus, nuclear volume and shape and nuclear to cell volume ratios. The true cell diagnosis for each cell was determined by a cytotechnologist and added to the feature set. Cross-validated classifiers were created using the technique of adaptive boosting that combines a sub-set of the features into a single score that best correlated with the binary cell diagnoses. Results: 3,256 normal cells or other normal objects and 1,843 breast cancer cells were studied. A classifier that distinguished normal cells from cancer cells was created that used the strongest 25 3D morphology features. The morphometric classifier had an area under the ROC curve that measured 0.9998. This classifier could be operated with near perfect specificity and a cancer cell sensitivity of 95%. Conclusions: Performance of automated, 3D morphometric classifiers for cancer cell detection suggests highly accurate CTC enumeration with implications for patient management and prediction of cancer progression.

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