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Circulating cancer cells

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https://www.readbyqxmd.com/read/28820176/5-hydroxymethylcytosine-signatures-in-cell-free-dna-provide-information-about-tumor-types-and-stages
#1
Chun-Xiao Song, Senlin Yin, Li Ma, Amanda Wheeler, Yu Chen, Yan Zhang, Bin Liu, Junjie Xiong, Weihan Zhang, Jiankun Hu, Zongguang Zhou, Biao Dong, Zhiqi Tian, Stefanie S Jeffrey, Mei-Sze Chua, Samuel So, Weimin Li, Yuquan Wei, Jiajie Diao, Dan Xie, Stephen R Quake
5-Hydroxymethylcytosine (5hmC) is an important mammalian DNA epigenetic modification that has been linked to gene regulation and cancer pathogenesis. Here we explored the diagnostic potential of 5hmC in circulating cell-free DNA (cfDNA) using a sensitive chemical labeling-based low-input shotgun sequencing approach. We sequenced cell-free 5hmC from 49 patients of seven different cancer types and found distinct features that could be used to predict cancer types and stages with high accuracy. Specifically, we discovered that lung cancer leads to a progressive global loss of 5hmC in cfDNA, whereas hepatocellular carcinoma and pancreatic cancer lead to disease-specific changes in the cell-free hydroxymethylome...
August 18, 2017: Cell Research
https://www.readbyqxmd.com/read/28819860/circulating-tumor-cells-markers-and-methodologies-for-enrichment-and-detection
#2
M J Alvarez Cubero, J A Lorente, I Robles-Fernandez, A Rodriguez-Martinez, J L Puche, M J Serrano
Cancer is a leading cause of disease worldwide; however, nowadays many points of its initiation processes are unknown. In this chapter, we are focusing on the role of liquid biopsies in cancer detection and progression. CTCs are one of the main components of liquid biopsies, they represent a subset of tumor cells that have acquired the ability to disseminate from the primary tumor and intravasate to the circulatory system. The greatest challenge in the detection of CTCs is their rarity in the blood. Human blood consists of white blood cells (5-10 × 10(6)/mL), red blood cells (5-9 × 10(9)/mL), and platelets (2...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819859/utilizing-matrigel-transwell-invasion-assay-to-detect-and-enumerate-circulating-tumor-cells
#3
Xingtong Liu, Xiangwei Wu
Metastasis is the cause of 90% of human cancer deaths. Circulating tumor cells (CTCs) in the peripheral blood and/or lymphatic vessels are cells shed from primary tumors and considered to be precursors of metastasis. Study of CTCs allows the serial monitoring of tumor progression and may provide predictive and prognostic biomarkers in clinic. Current CTC isolation and detection technologies encounter several challenges, including: heterogeneity of CTCs, low cell viability and/or high rate of contamination post-isolation, and the inability to distinguish viable/invasive from nonviable/nonfunctional CTCs, all of which can limit in vitro and in vivo characterization of CTCs...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819858/epcam-independent-enrichment-and-detection-of-viable-circulating-tumor-cells-using-the-epispot-assay
#4
Alexandra Soler, Laure Cayrefourcq, Martine Mazel, Catherine Alix-Panabières
Identification and characterization of circulating tumor cells (CTCs) in peripheral blood can provide information on the direction and the efficacy of treatments. Current techniques such as CellSearch(®) are limited in differentiating between apoptotic and viable CTCs. In contrast, the fluorescent EPISPOT assay allows for the identification of viable cells by detecting proteins secreted/released/shed by functional single epithelial cancer cells. In addition, as CTCs are rare events, it is required to combine the EPISPOT assay with an enrichment step...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819857/a-noninvasive-and-real-time-method-for-circulating-tumor-cell-detection-by-in-vivo-flow-cytometry
#5
Xunbin Wei, Jian Zhou, Xi Zhu, Xinrong Yang, Ping Yang, Qiyan Wang
The quantification of circulating tumor cells (CTCs) has been considered a potentially powerful tool in cancer diagnosis and prognosis, as CTCs have been shown to appear very early in cancer development. Great efforts have been made to develop methods that were less invasive and more sensitive to detect CTCs earlier. There is growing evidence that CTC clusters have greater metastatic potential than single CTCs. Therefore, the detection of CTC clusters is also important. This chapter is aimed to introduce a noninvasive technique for CTCs detection named in vivo flow cytometry (IVFC), which has been demonstrated to be capable of monitoring CTCs dynamics continuously...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819856/fiber-optic-array-scanning-technology-fast-for-detection-and-molecular-characterization-of-circulating-tumor-cells
#6
Zheng Ao, Xiaohe Liu
Circulating tumor cell (CTC) as an important component in "liquid biopsy" holds crucial clinical relevance in cancer prognosis, treatment efficiency evaluation, prediction and potentially early detection. Here, we present a Fiber-optic Array Scanning Technology (FAST) that enables antigen-agnostic, size-agnostic detection of CTC. By immunofluorescence staining detection of a combination of a panel of markers, FAST technology can be applied to detect rare CTC in non-small cell lung cancer (NSCLC) setting with high sensitivity and specificity...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819854/flow-cytometric-detection-of-circulating-tumor-cells-using-a-candidate-stem-cell-marker-p75-neurotrophin-receptor-p75ntr
#7
Tomoyuki Okumura, Tetsuji Yamaguchi, Toru Watanabe, Takuya Nagata, Yutaka Shimada
The most widely studied detection for circulating tumor cells (CTCs) in peripheral blood of cancer patients has been based on immunomagnetic enrichment using antibodies against epithelial cell adhesion molecule (EpCAM), which is overexpressed in epithelial cells. A neurotrophin receptor p75 (p75NTR) is expressed in a candidate stem cell fraction in esophageal squamous cell carcinoma (ESCC), which shows significantly higher colony formation, enhanced tumor formation in mice, along with strong expression of epithelial mesenchymal transition-related genes...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819852/enumeration-dielectrophoretic-capture-and-molecular-analysis-of-circulating-tumor-cells
#8
Stephanie S Yee, Erica L Carpenter
The identification of therapeutically targetable mutations in circulating tumor cells (CTCs) from cancer patient blood is increasingly used to personalize patient care. Here, we describe a novel approach for the enumeration, capture, and molecular analysis of CTCs from blood using an FDA-approved CTC enrichment and enumeration platform followed by dielectrophoretic capture and next-generation sequencing.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819847/negative-enrichment-and-isolation-of-circulating-tumor-cells-for-whole-genome-amplification
#9
Nisha Kanwar, Susan J Done
Circulating tumor cells (CTCs) are a rare population of cells found in the peripheral blood of patients with many types of cancer such as breast, prostate, colon, and lung cancers. Higher numbers of these cells in blood are associated with a poorer prognosis of patients. Genomic profiling of CTCs would help characterize markers specific for the identification of these cells in blood, and also define genomic alterations that give these cells a metastatic advantage over other cells in the primary tumor. Here, we describe an immunomagnetic method to enrich CTCs from the blood of patients with breast cancer, followed by single-cell laser capture microdissection to isolate single CTCs...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819846/filter-adapted-fluorescent-in-situ-hybridization-fa-fish-for-filtration-enriched-circulating-tumor-cells
#10
Marianne Oulhen, Emma Pailler, Vincent Faugeroux, Françoise Farace
Circulating tumor cells (CTCs) may represent an easily accessible source of tumor material to assess genetic aberrations such as gene-rearrangements or gene-amplifications and screen cancer patients eligible for targeted therapies. As the number of CTCs is a critical parameter to identify such biomarkers, we developed fluorescent in situ hybridization (FISH) for CTCs enriched on filters (filter-adapted-FISH, FA-FISH). Here, we describe the FA-FISH protocol, the combination of immunofluorescent staining (DAPI/CD45) and FA-FISH techniques, as well as the semi-automated microscopy method that we developed to improve the feasibility and reliability of FISH analyses in filtration-enriched CTC...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819845/automated-microfluidic-filtration-and-immunocytochemistry-detection-system-for-capture-and-enumeration-of-circulating-tumor-cells-and-other-rare-cell-populations-in-blood
#11
Michael Pugia, Mark Jesus M Magbanua, John W Park
Isolation by size using a filter membrane offers an antigen-independent method for capturing rare cells present in blood of cancer patients. Multiple cell types, including circulating tumor cells (CTCs), captured on the filter membrane can be simultaneously identified via immunocytochemistry (ICC) analysis of specific cellular biomarkers. Here, we describe an automated microfluidic filtration method combined with a liquid handling system for sequential ICC assays to detect and enumerate non-hematologic rare cells in blood...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819843/microfilter-based-capture-and-release-of-viable-circulating-tumor-cells
#12
Siddarth Rawal, Zheng Ao, Ram H Datar, Ashutosh Agarwal
Microfilters with slot-pore geometry can be used for size-based capture of circulating tumor cells (CTC) from the blood of cancer patients. The slot pore geometry reduces the shear stress that the cells would typically experience during filtration process and allows the cells to remain viable. The microfilter provides a platform capable of high CTC capture efficiency; however, the release of these cells from the filter following capture is nontrivial, possibly due to the strong nonspecific electrostatic adhesion of CTC to the microfilter surface...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819842/separable-bilayer-microfiltration-device-for-label-free-enrichment-of-viable-circulating-tumor-cells
#13
Sijie Hao, Merisa Nisic, Hongzhang He, Yu-Chong Tai, Si-Yang Zheng
Analysis of rare circulating tumor cells enriched from metastatic cancer patients yields critical information on disease progression, therapy response, and the mechanism of cancer metastasis. Here we describe in detail a label-free enrichment process of circulating tumor cells based on its unique physical properties (size and deformability). Viable circulating tumor cells can be successfully enriched and analyzed, or easily released for further characterization due to the novel separable two-layer design.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819841/microscale-laminar-vortices-for-high-purity-extraction-and-release-of-circulating-tumor-cells
#14
Soojung Claire Hur, James Che, Dino Di Carlo
Circulating tumor cells (CTCs) are disseminated tumor cells that reflect the tumors of origin and can provide a liquid biopsy that would potentially enable noninvasive tumor profiling, treatment monitoring, and identification of targeted treatments. Accurate and rapid purification of CTCs holds great potential to improve cancer care but the task remains technically challenging. Microfluidic isolation of CTCs within microscale vortices enables high-throughput and size-based purification of rare CTCs from bodily fluids...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819840/microfluidic-based-enrichment-and-retrieval-of-circulating-tumor-cells-for-rt-pcr-analysis
#15
Priya Gogoi, Saedeh Sepehri, Will Chow, Kalyan Handique, Yixin Wang
Molecular analysis of circulating tumor cells (CTCs) is hindered by low sensitivity and high level of background leukocytes of currently available CTC enrichment technologies. We have developed a novel device to enrich and retrieve CTCs from blood samples by using a microfluidic chip. The Celsee PREP100 device captures CTCs with high sensitivity and allows the captured CTCs to be retrieved for molecular analysis. It uses the microfluidic chip which has approximately 56,320 capture chambers. Based on differences in cell size and deformability, each chamber ensures that small blood escape while larger CTCs of varying sizes are trapped and isolated in the chambers...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819839/a-novel-microfluidic-device-for-isolation-of-circulating-tumor-cells-from-pancreatic-cancer-blood-samples
#16
Jose I Varillas, Kangfu Chen, Jinling Zhang, Thomas J George, Z Hugh Fan
Enumeration of circulating tumor cells (CTCs) can provide valuable prognostic information to guide cancer treatment as well as help monitor disease progression. Analysis of these rare malignant cells has the potential to further our understanding of cancer metastasis by gaining insights into CTC characteristics and properties. Microfluidics presents a unique platform to isolate and study CTCs. In this chapter, we describe the detailed procedures for the fabrication and use of a microfluidic device to detect CTCs from the blood of pancreatic cancer patients...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819838/microfluidic-separation-of-circulating-tumor-cells-based-on-size-and-deformability
#17
Emily S Park, Simon P Duffy, Hongshen Ma
Circulating tumor cells (CTCs) have been implicated as the seeds of cancer metastasis and therefore have the potential to provide significant prognostic and diagnostic values. Here, we describe a procedure for separating CTCs from whole blood based on size and deformability using the microfluidic ratchet device. This device leverages the ratcheting motion of single cells created as they are deformed through funnel-shaped constrictions using oscillatory flow in order to divert cells based on differences in size and deformability...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819837/microfluidic-capture-and-multiplex-immunofluorescence-of-circulating-tumor-cells-to-identify-cancer-of-origin
#18
Chian-Hui Lai, Ying-Chih Chang
Circulating tumor cells (CTCs) are an important biomarker and their analysis can be considered a form of "liquid biopsy." The purpose of this book chapter is to describe the use of the 4-channel CMx (cells captured in maximum) microfluidic chip, containing special micropatterns coated with an antibody-conjugated supported lipid bilayer (SLB) on its surface, to capture and isolate CTCs from the blood of cancer patients. Captured CTCs are subsequently released by an air foam to an immunofluorescence (IF) staining panel that enables further analysis, including the identification of the primary cancer source of the CTCs...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28819450/circulating-tumor-cells-moving-biological-insights-into-detection
#19
REVIEW
Lichan Chen, Ann M Bode, Zigang Dong
Circulating tumor cells (CTCs) have shown promising potential as liquid biopsies that facilitate early detection, prognosis, therapeutic target selection and monitoring treatment response. CTCs in most cancer patients are low in abundance and heterogeneous in morphological and phenotypic profiles, which complicate their enrichment and subsequent characterization. Several methodologies for CTC enrichment and characterization have been developed over the past few years. However, integrating recent advances in CTC biology into these methodologies and the selection of appropriate enrichment and characterization methods for specific applications are needed to improve the reliability of CTC biopsies...
2017: Theranostics
https://www.readbyqxmd.com/read/28819424/circulating-tumor-cells-and-microemboli-can-differentiate-malignant-and-benign-pulmonary-lesions
#20
Mario Mascalchi, Cristina Maddau, Lapo Sali, Elena Bertelli, Francesca Salvianti, Stefania Zuccherelli, Marzia Matucci, Alessandra Borgheresi, Claudio Raspanti, Monica Lanzetta, Massimo Falchini, Ernesto Mazza, Alessandra Vella, Michaela Luconi, Pamela Pinzani, Mario Pazzagli
The presence of circulating tumor cells (CTC) or microemboli (CTM) in the peripheral blood can theoretically anticipate malignancy of solid lesions in a variety of organs. We aimed to preliminarily assess this capability in patients with pulmonary lesions of suspected malignant nature. We used a cell-size filtration method (ScreenCell) and cytomorphometric criteria to detect CTC/CTM in a 3 mL sample of peripheral blood that was taken just before diagnostic percutaneous CT-guided fine needle aspiration (FNA) or core biopsy of the suspicious lung lesion...
2017: Journal of Cancer
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