Capture Of Circulating Tumor Cells Research Articles

Dual targeting negative enrichment strategy for highly sensitive and purity detection of CTCs

Circulating tumor cells (CTCs) have significant clinical value in early tumor detection, dynamic monitoring and immunotherapy. CTC detection stands out as a leading non-invasive approach for tumor diagnostics and therapeutics. However, the high heterogeneity of CTCs and the occurrence of epithelial-mesenchymal transition (EMT) during metastasis pose challenges to methods relying on EpCAM-positive enrichment. To address these limitations, a method based on negative enrichment of CTCs using specific leukocyte targets has been developed. In this study, aiming to overcome the low purity associated with immunomagnetic beads targeting solely the leukocyte common antigen CD45, we introduced CD66b-modified immunomagnetic beads. CD66b, a specific target for neutrophils with abundant residues, was chosen as a complementary approach. The process involved initial collection of nucleated cells from whole blood samples using density gradient centrifugation. Subsequently, magnetically labeled leukocytes were removed by magnetic field, enabling the capture of CTCs with higher sensitivity and purity while retaining their activity. Finally, we selected 20 clinical blood samples from patients with various cancers to validate the effectiveness of this strategy, providing a new generalized tool for the clinical detection of CTCs.

Abstract A003: Characterization of circulating tumor cells from patients with metastatic bladder cancer

Abstract Assessing the molecular profile of bladder tumors from patients with metastatic disease is crucial for implementing personalized treatment. Current standards are based on highly invasive biopsies. Alternatively, liquid biopsies of tumor cells, proteins, and nucleic acids allows longitudinal profiling in a less invasive manner. Circulating tumor cells (CTCs) isolated from blood are a key analyte of liquid biopsy. In this study, we developed a protein and transcriptomic co-analysis workflow for bladder cancer CTCs utilizing the graphene oxide (GO) microfluidic chip conjugated with antibodies against both EpCAM and EGFR to isolate CTCs from blood of patients with metastatic bladder cancer. Following CTC capture, protein and mRNA were analyzed using immunofluorescent staining and ion-torrent whole transcriptome sequencing. This is the first time that whole transcriptome sequencing was used in a downstream application for the GO microfluidic platform. We found that elevated CTC counts were significantly associated with patient disease status at the time of blood draw and that greater than 2 CTCs per mL were associated with shorter overall survival. Invasive markers, including EGFR, HER-2, CD31, and ADAM15, were detected in subpopulations of CTCs. Targeted whole transcriptome sequencing showed distinct expression profiles from patients with metastatic burden at the time of blood draw. In these patients, we found significant upregulation of several metastasis-related and chemotherapy-resistant genes demonstrating the capability of identifying tumor-related RNA signatures from circulating CTCs using immune-affinity GO chip-based assays. In addition, this workflow highlights the prognostic potential of CTCs in metastatic bladder cancer patients. Citation Format: Zeqi Niu, Molly Kozminski, Kathleen C. Day, Luke Broses, Marian L. Henderson, Sophia Merajver, Aaron Udager, Phillip L. Palmbos, Sunitha Nagrath, Mark L. Day. Characterization of circulating tumor cells from patients with metastatic bladder cancer [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2024 May 17-20; Charlotte, NC. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(10_Suppl):Abstract nr A003.

Development of a C3N4-based electrochemical sensing platform for the detection of circulating tumor cells in blood

This study presents a cancer diagnostics method through the development of a graphitic carbon nitride (g-C3N4)-based electrochemical sensor for detecting circulating tumor cells (CTCs) in blood. Utilizing the unique properties of g-C3N4 nanosheets combined with gold nanoparticles functionalized with DNA aptamers, the sensor exhibits remarkable sensitivity and selectivity in CTC detection. The aptamers specifically target cell surface biomarkers, enabling the precise capture and quantification of CTCs. The sensor's detection limit is an impressive 5 cells/mL, with a broad detection range from 10 to 105 cells/mL. This high-performance sensing platform not only achieves rapid and accurate quantification of CTCs but also demonstrates robustness against interference from other cell types, making it highly suitable for clinical applications. The optimized conditions for aptamer-functionalized nanoparticle concentration and incubation time ensure the sensor's efficacy, as validated in spiked blood samples. This work provides a practical and efficient approach for non-invasive cancer diagnostics, offering significant improvements over existing methods in terms of simplicity, sensitivity, and specificity for early detection and monitoring of cancer progression.

Circulating Tumor Cell Phenotype Detection and Epithelial-Mesenchymal Transition Tracking Based on Dual Biomarker Co-Recognition in an Integrated PDMS Chip.

Circulating tumor cells (CTCs) are widely considered as a reliable and promising class of markers in the field of liquid biopsy. As CTCs undergo epithelial-mesenchymal transition (EMT), phenotype detection of heterogeneous CTCs based on EMT markers is of great significance. In this report, an integrated analytical strategy that can simultaneously capture and differentially detect epithelial- and mesenchymal-expressed CTCs in bloods of non-small cell lung cancer (NSCLS) patients is proposed. First, a commercial biomimetic polycarbonate (PCTE) microfiltration membrane is employed as the capture interface for heterogenous CTCs. Meanwhile, differential detection of the captured CTCs is realized by preparing two distinct CdTe quantum dots (QDs) with red and green emissions, attached with EpCAM and Vimentin aptamers, respectively. For combined analysis, a polydimethylsiloxane (PDMS) chip with simple structure is designed, which integrates the membrane capture and QDs-based phenotype detection of CTCs. This chip not only implements the analysis of the number of CTCs down to 2 cellsmL-1, but enables EMT process tracking according to the specific signals of the two QDs. Finally, this method is successfully applied to inspect the correlations of numbers or proportions of heterogenous CTCs in 94 NSCLS patients with disease stage and whether there is distant metastasis.

A Detachable Magnetic Nanodevice for the Efficient Capture and Subtype Identification of Circulating Tumor Cells

AbstractThe effective capture and accurate identification of heterogeneous circulating tumor cells (CTCs) aid in guiding clinical diagnosis and personalized treatment of tumors. Herein, a core‐satellite‐sized magnetic separable nanodevice (denoted as MS‐RI) is developed for the capture and subtype identification of heterogeneous CTCs by identifying the protein targets of CTCs in vitro and imaging intracellular nucleic acid targets of CTCs in situ. Upon the separation of CTCs by MS‐RI using aptamers as capture elements, the programmatically dissociated satellite particles aim to internalize into the cells for simultaneous profiling of microRNA‐21 and microRNA‐141. By virtue of the validated magnetic separation of the core structure, the high affinity of the aptamer, and the high sensitivity of the recognition imaging (RI) module, MS‐RI allows the isolation and subtyping of CTCs in clinical samples from breast cancer patients. This strategy of combining extracellular recognition and intracellular imaging pioneers a new paradigm for the capture and precision subtyping of heterogeneous CTCs.

Size effect of dynamic bioactive magnetic particles on regulated isolation of tumor cells

Circulating tumor cells (CTCs) are critical biomarkers in cancer metastasis, yet their isolation from the bloodstream remains a challenging task due to their low abundance. This study introduces a method for the efficient isolation of CTCs, utilizing dynamic biointerface-based magnetic particles (DBMPs), with a particular emphasis on the role of particle size. We have developed three distinct sizes DBMPs specifically tailored for effective CTCs interaction, leveraging the unique properties of polydopamine (PDA) coatings and phenylboronic acid (PBA)-modified capture peptides through reversible catechol-boronate interactions. Comparative analysis of DBMPs of varying sizes was assessed under uniform experimental conditions for their capture and release efficiencies. The findings reveal a clear distinction: smaller particles (DBMP1 and DBMP2) achieved greater efficiency in capturing CTCs, whereas the larger particles (DBMP3) were more effective in releasing them. This difference is ascribed to the varied interaction dynamics between the particles and CTCs. Furthermore, we have fine-tuned DBMP3 to optimize CTC capture while still maintaining a high efficiency in cell release. The integration of these DBMPs into a magnetic isolation framework is poised to significantly enhance CTCs isolation methods, marking a pivotal advancement in cancer diagnostics and management.

Hybrid magnetic field system with helmholtz coils and magnets for real-time circulating tumor cell separation

Extensive research is currently underway to devise effective strategies for the isolation of circulating tumor cells (CTCs) from blood samples. The utilization of magnetic particles and magnet as a means for this separation process is garnering increased attention within this field. However, when utilizing fixed magnets to capture cancer cells with attached magnetic particles, the inability to control the magnetic field may result in cellular damage due to the strong magnetic field compressing the magnetic particles. In this study, we developed a hybrid system that leverages electromagnetic coils to modulate the strength of the magnetic field from the magnets within a microfluidic channel, enabling real-time control of cancer cells with attached magnetic particles.Through the strategic application of electromagnetic coils, we achieved precise control over the magnet’s position, thereby enabling real-time observation of the cancer cell within the microfluidic channel. Additionally, this approach facilitated the release of captured cancer cells from the magnet. The isolated cells could then be cultured to assess individualized drug responses, and evaluate their reactivity to the treatment. Through the implementation of this novel technique, we anticipate that the real-time capture and analysis of CTCs from blood samples will enable more efficient and effective diagnostics and treatment monitoring, ultimately improving patient outcomes.

Controllable Assembly of a Quantum Dot-Based Aptasensor Guided by CRISPR/Cas12a for Direct Measurement of Circulating Tumor Cells in Human Blood.

Accurate and sensitive analysis of circulating tumor cells (CTCs) in human blood provides a non-invasive approach for the evaluation of cancer metastasis and early cancer diagnosis. Herein, we demonstrate the controllable assembly of a quantum dot (QD)-based aptasensor guided by CRISPR/Cas12a for direct measurement of CTCs in human blood. We introduce a magnetic bead@activator/recognizer duplex core-shell structure to construct a multifunctional platform for the capture and direct detection of CTCs in human blood, without the need for additional CTC release and re-identification steps. Notably, the introduction of magnetic separation ensures that only a target-induced free activator can initiate the downstream catalysis, efficiently avoiding the undesired catalysis triggered by inappropriate recognition of the activator/recognizer duplex structure by crRNAs. This aptasensor achieves high CTC-capture efficiency (82.72%) and sensitive detection of CTCs with a limit of detection of 2 cells mL-1 in human blood, holding great promise for the liquid biopsy of cancers.

Multi-DNA-Modified Double-Network Hydrogel with Customized Microstructure: A Novel System for Living Circulating Tumor Cells Capture and Real-Time Detection.

The precise and effective isolation of living circulating tumor cells (CTCs) from peripheral blood, followed by their real-time monitoring, is crucial for diagnosing cancer patients. In this study, a cell-imprinted double-network (DN) hydrogel modified with circular multi-DNA (CMD), coined the CMD-imprinted hydrogel with fixed cells as templates (CMD-CIDH), was developed. The hydrogel featured a customized surface for proficient capture of viable CTCs and in situ real-time fluorescent detection without subsequent release. The customized surface, constructed using polyacrylamide/chitosan DN hydrogel as the matrix on the cell template, had a dense network structure, thereby ensuring excellent stability and a low degradation rate. Optimal capture efficiencies, recorded at 93 ± 3% for MCF-7 cells and 90 ± 2% for Hela cells, were achieved by grafting the CMD and adjusting the nodule size on the customized surface. The capture efficiency remained significantly high at 67 ± 11% in simulated breast cancer patient experiments even at a minimal concentration of 5 cells mL-1. Furthermore, CMD grafted onto the surface produced a potent fluorescence signature, enabling in situ real-time fluorescent detection of the target cell's growth state even in complex environments. The customized surface is highly efficient for screening CTCs in peripheral blood and has promising potential for setting up the CTCs culture.

Functionalized tetrahedral DNA frameworks for the capture of circulating tumor cells.

Identification and characterization of circulating tumor cells (CTCs) from blood samples of patients with cancer can help monitor parameters such as disease stage, disease progression and therapeutic efficiency. However, the sensitivity and specificity of current multivalent approaches used for CTC capture is limited by the lack of control over the ligands' position. In this Protocol Update, we describe DNA-tetrahedral frameworks anchored with aptamers that can be configured with user-defined spatial arrangements and stoichiometries. The modified tetrahedral DNA frameworks, termed 'n-simplexes', can be used as probes to specifically target receptor-ligand interactions on the cell membrane. Here, we describe the synthesis and use of n-simplexes that target the epithelial cell adhesion molecule expressed on the surface of CTCs. The characterization of the n-simplexes includes measuring the binding affinity to the membrane receptors as a result of the spatial arrangement and stoichiometry of the aptamers. We further detail the capture of CTCs from patient blood samples. The procedure for the preparation and characterization of n-simplexes requires 11.5 h, CTC capture from clinical samples and data processing requires ~5 h per six samples and the downstream analysis of captured cells typically requires 5.5 h. The protocol is suitable for users with basic expertise in molecular biology and handling of clinical samples.

Molecular Profiling of Circulating Tumour Cells and Circulating Tumour DNA: Complementary Insights from a Single Blood Sample Utilising the Parsortix® System.

The study of molecular drivers of cancer is an area of rapid growth and has led to the development of targeted treatments, significantly improving patient outcomes in many cancer types. The identification of actionable mutations informing targeted treatment strategies are now considered essential to the management of cancer. Traditionally, this information has been obtained through biomarker assessment of a tissue biopsy which is costly and can be associated with clinical complications and adverse events. In the last decade, blood-based liquid biopsy has emerged as a minimally invasive, fast, and cost-effective alternative, which is better suited to the requirement for longitudinal monitoring. Liquid biopsies allow for the concurrent study of multiple analytes, such as circulating tumour cells (CTCs) and circulating tumour DNA (ctDNA), from a single blood sample. Although ctDNA assays are commercially more advanced, there is an increasing awareness of the clinical significance of the transcriptome and proteome which can be analysed using CTCs. Herein, we review the literature in which the microfluidic, label-free Parsortix® system is utilised for CTC capture, harvest and analysis, alongside the analysis of ctDNA from a single blood sample. This detailed summary of the literature demonstrates how these two analytes can provide complementary disease information.

A novel electrochemiluminescent cytosensor using 3D multivalent aptamer recognition and covalent organic frameworks-associated DNA walker for highly efficient capture and detection of rare CTCs

Circulating tumor cells (CTCs) are regarded as promising biomarkers for early clinical diagnosis, prognosis evaluation, and treatment monitoring. However, their limited abundance in peripheral blood and phenotypic heterogeneity resulting from epithelial-mesenchymal transition (EMT) present significant challenges for accurate detection of CTCs. Herein, we developed an ultrasensitive and highly specific cytosensor to address these limitations. Firstly, to overcome the EMT property of CTCs, we designed a novel 3D multivalent aptamer recognition strategy (MARS), which not only contributed to highly efficient capture of heterogeneous CTCs, but also guaranteed detection sensitivity of the developed cytosensor. Meanwhile, we integrated an exonuclease III (Exo III)-mediated DNA walking signal amplification strategy into the electrochemiluminescence (ECL) cytosensor, enhancing its sensitivity in detecting rare CTCs. Furthermore, in combination with the excellent ECL performance of the luminescent nanomaterials (Au@COF-LZU1@Ru), the cytosensor exhibited a wide detection range from 8 to 1 × 105 cells/mL with a detection limit as low as 2 cells/mL. The analytical performance of the constructed cytosensor was validated through real sample analysis (AUC = 0.97), demonstrating its potential for clinical applications. The novel cytosensor offers an effective and reliable tool for the detection of rare CTCs, thereby paving the way for improved clinical significance.

Enumeration and Molecular Characterization of Circulating Tumor Cell Using an Epithelial Cell Adhesion Molecule/Vimentin/Epidermal Growth Factor Receptor Joint Capture System in Lung Cancer.

Detection rate and isolation yield of circulating tumor cells (CTCs) are low in lung cancer with approaches due to CTC invasiveness and heterogeneity. In this study, on the basis of the epithelial cell adhesion molecule (EpCAM) phenotype, markers of vimentin and epidermal growth factor receptor (EGFR) phenotype were added to jointly construct a precise and efficient CTC capture system for capture of lung cancer CTCs. A CTC capture system combined with EpCAM lipid magnetic bead (Ep-LMB)/vimentin lipid magnetic bead (Vi-LMB)/EGFR lipid magnetic bead (EG-LMB) was constructed, and its performance was tested. The amount of CTC captured in the blood of patients with lung cancer was detected by immunofluorescence identification and analyzed for clinical relevance. The constructed CTC capture system has low cytotoxicity. The capture efficiency of lung cancer cells in phosphate belanced solution (PBS) system was 95.48%. The capture efficiency in the blood simulation system is 94.55%. The average number of CTCs in the blood of patients with lung cancer was 9.73/2 mL. The quantity distribution of CTCs is significantly correlated with tumor staging and metastasis. The area under the curve (AUC) of CTCs for the diagnosis of lung cancer was 0.9994 (95% CI = 0.9981-1.000, P < .0001). The cutoff value was 4.5/2 mL. The sensitivity was 99.39%, and the specificity was 96.88%. The EpCAM/vimentin/EGFR combined capture system has feasibility and high sensitivity in the detection of lung cancer CTC typing, which can be used as an auxiliary diagnostic indicator for lung cancer and is expected to promote the clinical application of CTCs.

Metabolic Glycoengineering-Programmed Nondestructive Capture of Circulating Tumor Cells.

Circulating tumor cells (CTCs) are the "seeds" for malignant tumor metastasis, and they serve as an ideal target for minimally invasive tumor diagnosis. Abnormal glycolysis in tumor cells, characterized by glycometabolism disorder, has been reported as a universal phenomenon observed in various types of tumors. This provides a potential powerful tool for universal CTC capture. However, to the best of our knowledge, no metabolic glycoengineering-based CTC capture strategies have been reported. Here, we proposed a nondestructive CTC capture method based on metabolic glycoengineering and a nanotechnology-based proximity effect, allowing for highly specific, sensitive, and universal CTC capture. To achieve this goal, cells are first labeled with DNA tags through metabolic glycoengineering and then captured through a DNA tetrahedra-functionalized dual-tentacle magnetic nanodevice. Due to the difference in metabolic performance, only tumor cells are labeled with more densely packed DNA tags and captured through enhanced intermolecular interaction mediated by the proximity effect. In summary, we have constructed a versatile platform for nondestructive CTC capture, offering a novel perspective for the application of CTC liquid biopsy in tumor diagnosis and treatment.

Enhancing circulating tumor cell capture: Amin-functionalized bilayer graphene biosensing with integrated chip-level system for point-of-care testing

Current research on the detection of circulating tumor cells (CTCs) as a diagnostic strategy is still in its early stages. Consequently, there is a need for an efficient platform to capture and accurately detect CTCs with high sensitivity. This study presents the development of a bio-sensing platform that employs a planar-gated graphene field-effect transistor (GFET). This platform is designed for the detection of CTCs associated with nasopharyngeal carcinoma (NPC). In this approach, bilayer graphene is functionalized through a low-damage nitrogen plasma treatment, and then further modified with specific antibodies to enable a label-free detection of CTCs. The real-time electrical signal changes of the sensor were analyzed, demonstrating advantages in terms of rapid sensing and cost-effective on chip fabrication and instrument investment. The results showed that the low-damage nitrogen plasma effectively aminated the graphene surface, with the C–NH2 bonds accounting for approximately 62.8 % of the nitrogen content. The linear detection range of the sensor from 1 cell/15 μl to 1 cell/1000 μl, with a limit of detection (LoD) of 1 cell/15 μl(∼66/ml). The sensitivity of the sensor was determined to be 0.040 V/decade with a linear regression of 0.93 and with high sensing specificity in mixture cells environments. The CTCs capture density on aminated graphene was calculated to be 238.6 ± 29.8 cells/mm2. We also integrated this sensor chip into a portable system, including the sensing and signal processing unit enabling fast detection and close to point-of-care diagnostic tests. This study provides a promising platform for the rapid and accurate detection of nasopharyngeal carcinoma CTCs for early detection and post-treatment monitoring.

Dual-source powered nanomotors coupled with dual-targeting ligands for efficient capture and detection of CTCs in whole blood and in vivo tumor imaging

Circulating tumor cells (CTCs) are important biomarkers in cancer diagnosis. However, the specific labeling of CTCs with high capture efficiency in whole blood remains a problem. Herein, a dual-source-driven nanomotor coupled with dual-targeting ligands (CD@NM) was designed for efficient capture, specific imaging and quantitative detection of CTCs. In both water and biological fluid, CD@NMs moved autonomously under the propulsion of a magnetic field and H2O2 solution, which improved the capture efficiency of CTCs to 97.50 ± 2.38%. More importantly, specific labeling of CTCs was achieved by fluorescence quenching and recovery of fluorescent carbon dots modified on the CD@NMs. As a result, the CD@NMs exhibited efficient CTC capture, specific CTC imaging and recognition in whole blood. CD@NMs were also successfully deployed in the specific imaging of tumor tissues in vivo. On this basis, CD@NMs are expected to provide a new platform for tumor diagnosis both in vitro and in vivo.

Magnetically-activated, nanostructured cellulose for efficient capture of circulating tumor cells from the blood sample of head and neck cancer patients

In this report, the relative efficiency of cellulose nanocrystals (CNCs) and nanofibers (CNFs) to capture circulating tumor cells (CTCs) from the blood sample of head and neck cancer (HNC) patients was evaluated. Detection and enumeration of CTCs are critical for monitoring cancer progression. Both types of nanostructured cellulose were chemically modified with Epithelial Cell Adhesion Molecule (EpCAM) antibody and iron oxide nanoparticles. The EpCAM antibody facilitated the engagement of CTCs, promoting entrapment within the cellulose cage structure. Iron oxide nanoparticles, on the other hand, rendered the cages activatable via the use of a magnet for the capture and separation of entrapped CTCs. The efficiency of the network structures is shown in head and neck cancer (HNC) patients' blood samples. It was observed that the degree of chemical functionalization of hydroxyl groups located within the CNCs or CNFs with anti-EpCAM determined the efficiency of the system's interaction with CTCs. Further, our result indicated that inflexible scaffolds of nanocrystals interacted more efficiently with CTCs than that of the fibrous CNF scaffolds. Network structures derived from CNCs demonstrated comparable CTC capturing efficiency to commercial standard, OncoDiscover®. The output of the work will provide the chemical design principles of cellulosic materials intended for constructing affordable platforms for monitoring cancer progression in ‘real time’.

Circulating Tumor Cells Adhesion: Application in Biosensors.

The early and non-invasive diagnosis of tumor diseases has been widely investigated by the scientific community focusing on the development of sensors/biomarkers that act as a way of recognizing the adhesion of circulating tumor cells (CTCs). As a challenge in this area, strategies for CTCs capture and enrichment currently require improvements in the sensors/biomarker's selectivity. This can be achieved by understanding the biological recognition factors for different cancer cell lines and also by understanding the interaction between surface parameters and the affinity between macromolecules and the cell surface. To overcome some of these concerns, electrochemical sensors have been used as precise, fast-response, and low-cost transduction platforms for application in cytosensors. Additionally, distinct materials, geometries, and technologies have been investigated to improve the sensitivity and specificity properties of the support electrode that will transform biochemical events into electrical signals. This review identifies novel approaches regarding the application of different specific biomarkers (CD44, Integrins, and EpCAm) for capturing CTCs. These biomarkers can be applied in electrochemical biosensors as a cytodetection strategy for diagnosis of cancerous diseases.

A microfluidic device for enhanced capture and high activity release of heterogeneous CTCs from whole blood

Circulating tumor cells (CTCs) are tumor cells that spontaneously detach from the primary focus, and early detection and characterization of CTCs is vital for cancer diagnosis and appropriate treatment. Current methods commonly use EpCAM to capture CTCs, but this results in a loss of information on other CTC subsets (EpCAM-negative cells) due to the heterogeneity of CTCs. Here, we report a novel microfluidic device that integrates the capture and release of heterogeneous CTCs directly from whole blood. A spiral chip was designed for the separation of differently sized cells, and larger CTCs were effectively separated from smaller blood cells with a 98% recovery rate. CD146-containing magnetic beads were used to complement the EpCAM-based CTC capture methods, and the capture efficiency of Fe3O4@Gelatin@CD146/EpCAM increased by 20% over Fe3O4@Gelatin@EpCAM. Finally, MMP-9 was employed to release CTCs with high efficiency and less damage by degrading gelatins on the surface of Fe3O4. The established method was successfully applied to CTC capture and release in a simulated patient's whole blood. The developed method achieved enhanced capture and high activity release of heterogeneous CTCs with less interference by blood cells, which contributes to the early detection and clinical downstream analysis of CTCs.

Automated Capture and Analysis of Circulating Tumor Cells in Pediatric, Adolescent and Young Adult Patients with Central Nervous System Tumors.

Tumors of the central nervous system (CNS) are the most common and lethal childhood malignancy. Detection of residual disease and longitudinal monitoring of treatment response in patients are challenging and rely on serial imaging. This current standard of care fails to detect microscopic disease or provide molecular characteristics of residual tumors. As such, there is dire need for minimally invasive liquid biopsy techniques. We have previously shown the high specificity of using cell surface vimentin (CSV) to identify circulating tumor cells (CTCs) from patients bearing various types of cancers. Here, we describe the first report of CTCs captured from peripheral blood samples in 58 pediatric CNS tumor patients. In this study, we used a CSV-coated cell capture chip, the Abnova CytoQuest automated CTC isolation system, to boost the CTC capture from pediatric patients with CNS tumors. We successfully isolated CTCs in six glioma patients using immunostaining of histone H3 lysine27-to-methionine (H3K27M) mutations which are highly expressed by this tumor. We show that CSV is a viable marker for CNS CTC isolation and that this is a feasible method for detecting microscopic disease. Larger-scale studies focusing on CTCs in pediatric CNS tumors to explore their diagnostic and prognostic value are warranted.