Cholangiocarcinoma (CCA), also known as bile duct cancer, is a devastating malignancy primarily affecting the biliary tract.

To assess their performance in clinical diagnosis and monitoring of CCA, plasma methylation and circulating tumor cells were detected.

Plasma samples were collected from Hubei Cancer Hospital (n = 156). Plasma DNA was tested to detect SHOX2, HOXA9, SEPTIN9, and RASSF1A methylation using TaqMan PCR. Circulating tumor cells (CTCs) were detected in the peripheral blood of patients using the United States Food and Drug Administration-approved cell search system before and after clinical therapy. The CCA diagnostic value was estimated using the area under the curve. The independent prognosis risk factors for patients with CCA were estimated using Cox and logistic regression analyses.

The sensitivity and specificity of the four DNA plasma methylations exhibited 64.74% sensitivity and 93.88% specificity for detecting CCA. The receiver operating characteristic curve of the combined value for CCA diagnosis in plasma was 0.828 ± 0.032. RASSF1A plasma methylation was related to the prognosis of patients with CCA. We determined the prognostic hazard ratio for CCA using CTC count, tumor stage, methylation, and carbohydrate antigen 19-9 levels as key factors. Our overall survival nomogram achieved a C-index of 0.705 (0.605-0.805).

SHOX2, HOXA9, SEPTIN9, and RASSF1A plasma methylation demonstrated increased sensitivity for diagnosing CCA. RASSF1A plasma methylation and CTCs were valuable predictors to assess CCA prognosis and recurrence.

• Cholangiocarcinoma
• Methylation
• Circulating cancer cells
• Diagnosis
• Prognosis

• Biomarkers
• Diagnosis
• Gastric cancer
• Liquid biopsies
• Prognosis

• Circulating tumor cell (CTC)
• Flexible graphene-based biosensor
• Ovarian cancer

• Science and Technology Plan Projects from Sichuan

The presence of cancer cells clusters is a frequent event capable of increasing their aptitude to survive in the bloodstream. Consistently, clusters ranging from 2–50 cancer cells are detected in about 50% of patients with metastatic cancers, including colorectal carcinoma. Although a deepened analysis of clusters might certainly offer new insights into the complexity of metastatic cascade, research in this field has come to a halt, since most circulating tumor cells isolation techniques are not compatible with large-sized clusters isolation. In the present study, we describe a sequential method to simultaneously isolate single and clustered circulating tumor cells from a single blood draw, opening new scenarios for an ever more precise characterization of colorectal cancer metastatic cascade.

Circulating tumor cells (CTCs) detach from a primary tumor or its metastases and circulate in the bloodstream. The vast majority of CTCs are deemed to die into the bloodstream, with only few cells representing viable metastatic precursors. Particularly, single epithelial CTCs do not survive long in the circulation due to the loss of adhesion-dependent survival signals. In metastatic colorectal cancer, the generation of large CTC clusters is a very frequent occurrence, able to increase the aptitude of CTCs to survive in the bloodstream. Although a deepened analysis of large-sized CTC clusters might certainly offer new insights into the complexity of the metastatic cascade, most CTC isolation techniques are unfortunately not compatible with large-sized CTC clusters isolation. The inappropriateness of standard CTC isolation devices for large clusters isolation and the scarce availability of detection methods able to specifically isolate and characterize both single CTCs and CTC clusters finally prevented in-depth studies on the prognostic and predictive value of clusters in clinical practice, unlike that which has been described for single CTCs. In the present study, we validated a new sequential filtration method for the simultaneous isolation of large CTC clusters and single CTCs in patients with metastatic colorectal cancer at failure of first-line treatments. The new method might allow differential downstream analyses for single and clustered CTCs starting from a single blood draw, opening new scenarios for an ever more precise characterization of colorectal cancer metastatic cascade.

• biofluids
• biomarkers
• haematological malignancies
• leukaemia
• lymphoma
• multiple myeloma
• proteomics

• Biomarkers, Tumor/metabolism
• Body Fluids/metabolism
• Hematologic Neoplasms/diagnosis
• Hematologic Neoplasms/metabolism
• Humans
• Neoplasm Proteins/metabolism
• Proteomics

• cell biology
• molecular biology

• Actomyosin/metabolism
• Caco-2 Cells
• Cell Movement/physiology
• Cell Polarity
• Cell Shape
• Endosomes/metabolism
• Epithelial Cell Adhesion Molecule/metabolism
• Epithelial Cells/metabolism
• HeLa Cells
• Humans
• Myosin Type II/metabolism
• Stress Fibers/metabolism
• rhoA GTP-Binding Protein/metabolism

• Biomarker
• Cancer
• Drug screening
• Microfluidic chip
• Nanoparticle
• Preclinical model

• EpCAM
• black phosphorus
• circulating tumor cells
• intravenous catheters
• photothermal therapy

• Shenzhen University
• National Natural Science Foundation of China
• China Postdoctoral Science Foundation

• cancer immunotherapy
• immunopeptidomics
• mass spectrometry
• personalized drug therapy
• pharmacoepigenomics
• pharmacogenomics (PGx)
• pharmacomicrobiomics
• precision medicine
• therapeutic drug monitoring (TDM)

• Anti-Bacterial Agents/pharmacology
• Antineoplastic Agents
• Biomarkers, Pharmacological/analysis
• DNA Methylation/drug effects
• Drug Monitoring/methods
• Drug Therapy/methods
• Histones/metabolism
• Humans
• Liquid Biopsy
• Mass Spectrometry/methods
• Pharmacogenomic Testing/methods
• Precision Medicine/methods

Circulating tumor cells (CTCs) carry a wealth of information on primary and metastatic tumors critical for enhancing the understanding of the occurrence, progression and metastasis of non-small cell lung cancer (NSCLC). However, the low sensitivity of traditional tumor detection methods limits the application of CTCs in the treatment and disease surveillance of NSCLC. Therefore, CTCs isolation and detection with high sensitivity is highly desired especially for NSCLC patients, which is significant because of high occurrence and mortality. While it is very challenging because of the lower expression of CTC positive biomarkers such as epithelial cell adhesion molecules and cytokeratins (EpCAM and CKs), herein we report a method based on peptide-functionalized magnetic nanoparticles with high CTC capture efficiency, which demonstrates superiority in NSCLC clinical applications.

For analysis and comparison of the peptide-functionalized magnetic nanoparticles (TumorFisher, Nanopep Corp.) and the antibody-modified magnetic beads (CellSearch, Janssen Diagnostics, LLC), two NSCLC cell lines, A549 and NCI-H1975 were chosen to measure the binding affinity and capture efficiency. In order to compare the effect of the clinical application of these two detection systems, 7 early stage patients with NSCLC were enrolled in this study. To further explore the clinical utility of CTC counting in different stages, 81 NSCLC patients in stage I–IV were enrolled for CTC enumeration and statistical analysis.

The binding affinities of the recognition peptide to A549 and NCI-H1975 are 76.7%±11.0% and 70.1%±4.8%, respectively, which is similar with the positive control group (anti-EpCAM antibodies). CTCs were captured in 5/7 (71.4%) of early stage NSCLC patients with NSCLC in TumorFisher system, which is higher than CellSearch, and the false negative of TumorFisher is much lower than CellSearch. In a larger clinical cohort, the CTC numbers of NSCLC patients varied in different stages and the overall detection rate of TumorFisher was 59/81 (72.8%), with the similar proportion in stage I (21/29, 72.4%), II (17/22, 77.3%) and III (16/21, 76.2%).

Highly efficient CTC isolation technique based on peptide-magnetic nanoparticles was firstly applied in NSCLC patients. Compared with the antibody-based the technique, the higher CTC detection rates (71.4%) in NSCLC patient blood samples were demonstrated for the patients in different stages, I–IV, especially in early stages. This indicates the feasibility of the clinical utility of this new technique in early stage screening, prognosis and therapy evaluation of NSCLC.

• circulating tumor cell (CTC)
• liquid biopsy
• nanoparticle
• non-small cell lung cancer (NSCLC)
• peptide (Pep)

Patients with operable breast cancer have a better prognosis for recovery. However, once distant organ metastasis occurs, the chance of a long-term survival or a cure is limited. The collection and counting of circulating tumor cells (CTCs) by reliable detection techniques are of increasing importance in the diagnosis of early metastasis and prognosis of disease progression. Isolation by size of epithelial tumor cells (ISET) has the advantage of simplicity of operation and high homogeneity. It is practical for large-scale clinical detection showing cell abundance. The value of ISET in the detection of circulating breast cancers in the blood has not been determined. The purpose of this study is to explore the feasibility of applying ISET to detect CTCs by determining the detection rate of ISET in operable breast cancer and by evaluating the correlation between detection rate, cell count and clinical factors such as molecular typing and pathological staging.

The experiment included 193 breast cancer patients who were diagnosed by core needle biopsy before the operation. 10 mL of venous blood was collected from the patients preoperatively, and CTCs in their blood samples were counted and analyzed by ISET.

Patients were divided into groups according to pathology and immunohistochemistry. The overall detection rate of CTCs by ISET was 41.24%. The detection rate, the number of overall CTCs and the average number of CTCs in each group were analyzed individually. No significant differences were observed between the different groups.

Although ISET has a relatively good detection rate for circulating breast cancer cells, it fails to provide more information on pathological staging, molecular classification and so forth.

• Isolation by size of epithelial tumor cells (ISET)
• breast cancer
• circulating tumor cells (CTCs)

• Cancer nanomedicine
• Clinical imaging
• Combination therapy
• Gene therapy
• Immune therapy
• Nanodiagnostics
• Nanotechnology
• Theranostics

• Breast cancer
• CTC
• death rate
• primary
• prognosis
• relapse rate.

• circulating tumor cells
• epithelial cell adhesion molecule
• folate receptor alpha
• ovarian cancer