Current Circulating Tumor Cells Research Articles

Microfluidic System Consisting of a Magnetic 3D-Printed Microchannel Filter for Isolation and Enrichment of Circulating Tumor Cells Targeted by Anti-HER2/MOF@Ferrite Core-Shell Nanostructures: A Theranostic CTC Dialysis System.

Low number of circulating tumor cells (CTCs) in the blood samples and time-consuming properties of the current CTC isolation methods for processing a small volume of blood are the biggest obstacles to CTC usage in practice. Therefore, we aimed to design a CTC dialysis system with the ability to process cancer patients' whole blood within a reasonable time. Two strategies were employed for developing this dialysis setup, including (i) synthesizing novel in situ core-shell Cu ferrites consisting of the Cu-CuFe2O4 core and the MIL-88A shell, which are targeted by the anti-HER2 antibody for the efficient targeting and trapping of CTCs; and (ii) fabricating a microfluidic system containing a three-dimensional (3D)-printed microchannel filter composed of a polycaprolactone/Fe3O4 nanoparticle composite with pore diameter less than 200 μm on which a high-voltage magnetic field is focused to enrich and isolate the magnetic nanoparticle-targeted CTCs from a large volume of blood. The system was assessed in different aspects including capturing the efficacy of the magnetic nanoparticles, CTC enrichment and isolation from large volumes of human blood, side effects on blood cells, and the viability of CTCs after isolation for further analysis. Under the optimized conditions, the CTC dialysis system exhibited more than 80% efficacy in the isolation of CTCs from blood samples. The isolated CTCs were viable and were able to proliferate. Moreover, the CTC dialysis system was safe and did not cause side effects on normal blood cells. Taken together, the designed CTC dialysis system can process a high volume of blood for efficient dual diagnostic and therapeutic purposes.

A combination of circulating tumor cell (CTC) and cancer stem cell (CSC) markers to predict the prognosis of breast cancer.

e13007 Background: CTCs are tumor cells that shed from either primary tumor or metastatic foci and can provide more information about the tumor's biological entity than circulating tumor DNA (ctDNA), or exosomes do. The current FDA-approved CTC assay misses the CSC phenotypes, which are pivotal in driving tumorigenesis, metastasis, resistance to treatment, and cancer recurrence. Therefore, we combine CTC and CSC markers to determine the prognosis. We hypothesized that the CSC markers on CTC correlate with poorer outcomes. Methods: Peripheral blood samples of qualified breast cancer patients were collected at diagnosis and every 6 months during follow-up. EpCAM, MUC-1, and HER2 are used as CTC markers, while TWIST, AKT, PI3K, and ALDH-1 are used to detect CSC. According to the expression level of the above markers, patients were categorized into four groups: NN negative with both CTC and CSC markers, NP expressing CSC markers, PN expressing CTC markers, and PP positive with both CTC and CSC markers. The progression-free survival (PFS) and overall survival (OS) of all four groups were calculated by Kaplan-Meier model from the date of surgery until the disease progression and death/the last follow-up of the patients, respectively. Results: We screened 100 patients aged 18 years or older with confirmed stage II to stage IV breast cancer and selected 81 patients for the study. Fifteen patients dropped out of the study due to loss of follow-up or incomplete clinical information leaving 66 evaluable patients. There are 55 Caucasians (83%), 5 African Americans (7.5%), 5 Hispanics (7.5%), and 1 Asian (1.5%). The median age of the patients is 52.5 years old (29-81), and the median follow-up is 22.6 months (2.5-65). Most patients have stage III disease (60 and 91%) followed by 4 stage IV (6%) and 2 Stage II (3%). The median time between the first sample date and surgery is 172 days (-148 to 270). The NN group has the most patients (41 and 62%) compared to NP group (16 and 24%) and PN group (6 and 9%). The PP group has the least number of patients (2 and 3%) and is not observed in patients with positive hormonal receptors (HR). The PN group has a higher ratio of HR-HER2+ patients than NP group does. The HR and HER2 statuses were checked in the four groups. Interestingly, the NN and NP groups are mostly HR+HER2- patients, while the PN group is mainly HR- patients regardless of the HER2 status. The PP group only has HR- patients. The PFS curves start to flatten at 12 months and separate among the four groups, with the highest PFS percentage in the NP (81.2%) and NN (77.8%) followed by the PN (62.5%) and PP (50%) group. The OS percentage is 70% in NN, 66% in PN, 60% in NP, and 50% in PP, similar to the PFS pattern. Conclusions: Checking both CTC and CSC markers help triage the prognosis of breast cancer patients. CTC and CSC markers observed in breast cancer patients with different HR or HER2 status can be further investigated for potential correlations.

Effective Circulating Tumor Cell Isolation Using Epithelial and Mesenchymal Markers in Prostate and Pancreatic Cancer Patients

Circulating tumor cells (CTCs) display antigenic heterogeneity between epithelial and mesenchymal phenotypes. However, most current CTC isolation methods rely on EpCAM (epithelial cell adhesion molecule) antibodies. This study introduces a more efficient CTC isolation technique utilizing both EpCAM and vimentin (mesenchymal cell marker) antibodies, alongside a lateral magnetophoretic microseparator. The effectiveness of this approach was assessed by isolating CTCs from prostate (n = 17) and pancreatic (n = 5) cancer patients using EpCAM alone, vimentin alone, and both antibodies together. Prostate cancer patients showed an average of 13.29, 11.13, and 27.95 CTCs/mL isolated using EpCAM alone, vimentin alone, and both antibodies, respectively. For pancreatic cancer patients, the averages were 1.50, 3.44, and 10.82 CTCs/mL with EpCAM alone, vimentin alone, and both antibodies, respectively. Combining antibodies more than doubled CTC isolation compared to single antibodies. Interestingly, EpCAM antibodies were more effective for localized prostate cancer, while vimentin antibodies excelled in metastatic prostate cancer isolation. Moreover, vimentin antibodies outperformed EpCAM antibodies for all pancreatic cancer patients. These results highlight that using both epithelial and mesenchymal antibodies with the lateral magnetophoretic microseparator significantly enhances CTC isolation efficiency, and that antibody choice may vary depending on cancer type and stage.

Baits-trap chip for accurate and ultrasensitive capture of living circulating tumor cells.

Accurate analysis of living circulating tumor cells (CTCs) plays a crucial role in cancer diagnosis and prognosis evaluation. However, it is still challenging to develop a facile method for accurate, sensitive, and broad-spectrum isolation of living CTCs. Herein, inspired by the filopodia-extending behavior and clustered surface-biomarker of living CTCs, we present a unique baits-trap chip to achieve accurate and ultrasensitive capture of living CTCs from peripheral blood. The baits-trap chip is designed with the integration of nanocage (NCage) structure and branched aptamers. The NCage structure could "trap" the extended filopodia of living CTCs and resist the adhesion of filopodia-inhibited apoptotic cells, thus realizing the accurate capture (∼95% accuracy) of living CTCs independent of complex instruments. Using an in-situ rolling circle amplification (RCA) method, branched aptamers were easily modified onto the NCage structure, and served as "baits" to enhance the multi-interactions between CTC biomarker and chips, leading to ultrasensitive (99%) and reversible cell capture performance. The baits-trap chip successfully detects living CTCs in broad-spectrum cancer patients and achieves high diagnostic sensitivity (100%) and specificity (86%) of early prostate cancer. Therefore, our baits-trap chip provides a facile, accurate, and ultrasensitive strategy for living CTC isolation in clinical. STATEMENT OF SIGNIFICANCE: A unique baits-trap chip integrated with precise nanocage structure and branched aptamers was developed for the accurate and ultrasensitive capture of living CTCs. Compared with the current CTC isolation methods that are unable to distinguish CTC viability, the nanocage structure could not only "trap" the extended-filopodia of living CTCs, but also resist the adhesion of filopodia-inhibited apoptotic cells, thus realizing the accurate capture of living CTCs. Additionally, benefiting from the "baits-trap" synergistic effects generated by aptamer modification and nanocage structure, our chip achieved ultrasensitive, reversible capture of living CTCs. Moreover, this work provided a facile strategy for living CTC isolation from the blood of patients with early-stage and advanced cancer, exhibiting high consistency with the pathological diagnosis.

Numerical investigation of centrifugal passive cell separation in three types of serpentine microchannels and comparison with fixed platform

Cell separation plays a crucial role in diagnosing and improving a wide range of diseases, such as cancer, which is a severe reason for the death of people in the current decades. Circulating tumor cells (CTCs) inside the blood can be separated from the other whole blood cells to detect early cancer. Inertial methods are more superficial and cost less than the current CTC separation methods. These methods also have great potential in high-throughput separation in lab-on-a-chip (LOC) and lab-on-a-disk (LOD) devices because of secondary flow generation, especially in serpentine-shaped microchannels. The present study considers a continuous process for separating CTCs from the blood sample. This process is utilized in LOD devices, and this platform is also compared with the LOC platform. Moreover, two common types of different serpentine-shaped channels (simple and curved) and a new Omega channel are compared. It should be noted that a direct numerical simulation (DNS) method is used for calculating lift force in the serpentine-shaped channels. The effect of fluid velocity on WBC and CTC separation efficiency in the three mentioned different geometries are investigated in both fixed and rotational platforms, then the best results for each of them are presented. Finally, the best results belonged to the simple serpentine between the three investigated channels, with a separation percentage of 100. Moreover, in comparing the two mentioned platforms, LOD showed a more efficient application in cell separation. In addition, the Omega channel is investigated as a novel serpentine geometry. According to the results, it can be used for particle focusing applications thanks to its 100 percent efficiency at 0.5 m/s in both LOD and LOC platforms.

Utilization of Circulating Tumor Cells in the Management of Solid Tumors.

Circulating tumor cells (CTCs) are tumor cells shed from the primary tumor into circulation, with clusters of CTCs responsible for cancer metastases. CTC detection and isolation from the bloodstream are based on properties distinguishing CTCs from normal blood cells. Current CTC detection techniques can be divided into two main categories: label dependent, which depends upon antibodies that selectively bind cell surface antigens present on CTCs, or label-independent detection, which is detection based on the size, deformability, and biophysical properties of CTCs. CTCs may play significant roles in cancer screening, diagnosis, treatment navigation, including prognostication and precision medicine, and surveillance. In cancer screening, capturing and evaluating CTCs from peripheral blood could be a strategy to detect cancer at its earliest stage. Cancer diagnosis using liquid biopsy could also have tremendous benefits. Full utilization of CTCs in the clinical management of malignancies may be feasible in the near future; however, several challenges still exist. CTC assays currently lack adequate sensitivity, especially in early-stage solid malignancies, due to low numbers of detectable CTCs. As assays improve and more trials evaluate the clinical utility of CTC detection in guiding therapies, we anticipate increased use in cancer management.

A novel whole blood purifier for efficient capture and separation of circulating tumor cells

Circulating tumor cells (CTCs) as important biomarkers for noninvasive clinical diagnosis and prognostic evaluation are significant in predicting the overall survival and progression-free survival of cancer patients. However, the current typical CTCs separation and enrichment techniques were limited to a single collection of small-volume blood samples, which was inadequate to comprehensively profile the distribution of CTCs in the systemic blood. In addition, those techniques cannot reduce metastasis of CTCs unless adjuvant chemotherapy. Herein, inspired by hemodialysis, we designed a whole blood purifier (WBP) composed of a functionalized special spiral-like glass tube modified by anti-epithelial cell adhesion molecule (anti-EpCAM). The WBP allowed real-time capture, enrichment and removal of CTCs from systemic blood circulation, and the purified blood was immediately returned to the body. Furthermore, the WBP did not cause any organic damages in vivo. This approach achieves the high accuracy of liquid biopsy technology and is expected to become an effective clinical adjuvant therapy for tumor metastasis.

Reversible capture and release of circulating tumor cells on a three‐dimensional conductive interface to improve cell purity for gene mutation analysis

AbstractRare circulating tumor cells (CTCs) are always harvested with large numbers of white blood cells (WBCs) by current CTC isolation techniques, which influences the accuracy of CTC‐related gene and protein analysis. Therefore, it is very urgent to develop a method for efficient isolation of CTCs with high purity. In this work, we fabricated a reversibly assembled interface by layer‐by‐layer assembly of biotinylated poly‐(L‐lysine‐graft‐ethylene glycol) (PLL‐g‐PEG‐biotin), streptavidin, and CTC‐specific antibody on a three‐dimensional conductive scaffold, and further embedded it into a customized microchip for CTC capture. The assembled multilayers increased the roughness of the scaffold, which improved the capture efficiency. Importantly, the PLL‐g‐PEG‐biotin covalently coupled with CTCs could be detached from the scaffold by electrostatic repulsion, and the released CTCs could be recaptured on the conductive scaffold via electrostatic attraction while the WBCs were continuously drifted away. This reversible capture and release strategy significantly improved the purity of CTCs, and pure CTCs were obtained from cancer patients’ blood samples for gene mutation assay, which will be of great significance for assisting cancer diagnosis and treatment in a noninvasive way.

Bioinspired nano-plate-coral platform enabled efficient detection of circulating tumor cells via the synergistic capture of multivalent aptamer and tumor cell membrane

Circulating tumor cells (CTCs) offer rich information for early disease diagnosis and therapy evaluation. However, the limited sensitivity, binding affinity, and stability of current monovalent recognition-based CTCs detection techniques remain a challenge for extending their applications. Inspired by the highly efficient predation manner of plate corals, we firstly introduce an efficient and sensitive biomimetic CTCs recognition platform based on the conjugation of multivalent aptamer onto tumor cell membrane-coated magnetic graphene oxide to form a plate coral-like CTCs capture nanoprobe (MNPA-TCMMGO). In this method, the tumor cell membrane was employed to provide a biomimetic homologous fluidic interface for targeting homologous tumor cells. At the same time, multivalent aptamers were used as capture probes, which greatly enhanced the binding affinity and association probability between aptamer and target cells via cooperative multivalent effect. The unique features (robustness, high binding affinity and specificity, and biocompatibility) of MNPA-TCMMGO allow efficient, sensitive, and specific capture of rare tumor cells from biological samples. More importantly, the captured cells could maintain good viability, which is crucial for downstream analysis. Therefore, our developed biomimetic approach offers a new way to address the limitations of current CTCs detection methods and presents considerable potential for clinical cancer diagnostics.

Advances in the Biology, Detection Techniques, and Clinical Applications of Circulating Tumor Cells.

Circulating tumor cells (CTCs) play a crucial role in tumor recurrence and metastasis, and their early detection has shown remarkable benefits in clinical theranostics. However, CTCs are extremely rare, thus detecting them in the blood is very challenging. New CTC detection techniques are continuously being developed, enabling deeper analysis of CTC biology and potential clinical application. This article reviews current CTC detection techniques and their clinical application. CTCs have provided, and will continue to provide, important insights into the process of metastasis, which could lead to development of new therapies for different cancers.

2D MOF Nanosensor-Integrated Digital Droplet Microfluidic Flow Cytometry for In Situ Detection of Multiple miRNAs in Single CTC Cells.

Current circulating tumor cells (CTCs) detection strategies based on surface epithelial markers suffer from low specificity in distinguishing between CTCs and epithelial cells in hematopoietic cell population. Tumor-associated miRNAs within CTCs are emerging as new biomarkers due to their high correlation with tumor development and progress. However, in-situ simultaneous analysis of multiple miRNAs in single CTC cell is still challenging. To overcome this limitation, a digital droplet microfluidic flow cytometry based on biofunctionalized 2D metal-organic framework nanosensor (Nano-DMFC) is developed for in situ detection of dual miRNAs simultaneously in single living breast cancer cells. Here, 2D MOF-based fluorescent resonance energy transfer (FRET) nanosensors are established by conjugating dual-color fluorescence dye-labeled DNA probes on MOF nanosheet surface. In the Nano-DMFC, 2D MOF-based nanoprobes are precisely microinjected into each single-cell encapsulated droplets to achieve dual miRNA characterization in single cancer cell. This Nano-DMFC platform successfully detects dual miRNAs at single-cell resolution in 10 mixed positive MCF-7 cells out of 10 000 negative epithelial cells in serum biomimic samples. Moreover, this Nano-DMFC platform shows good reproductivity in the recovery experiment of spiked blood samples, which demonstrate the high potential for CTC-based cancer early diagnosis and prognosis.

DETECT: Development of Technologies for Early HCC Detection

DETECT: Development of Technologies for Early HCC Detection

A chimeric virus-based probe unambiguously detects live circulating tumor cells with high specificity and sensitivity

The current methods for detecting circulating tumor cells (CTCs) suffer from several drawbacks. We report a novel method that is based on a chimeric virus probe and can detect CTCs with extremely high specificity and sensitivity. Moreover, it exclusively detects live CTCs, and its detection efficacy is not impacted by the variation of epithelial cell adhesion molecule (EpCAM) expression. The chimeric virus probe is composed of a capsid from human papillomavirus that provides the detection with high specificity and an SV40-based genome that can amplify extensively inside CTCs and, hence, endows the detection with high sensitivity. Furthermore, different marker genes can be incorporated into the probe to provide detection with versatility. These unique capabilities will likely improve the validity and utility of this CTC detection in several clinical applications, which is one of the drawbacks suffered by many of the current CTC detection methods.

Cell-Released Magnetic Vesicles Capturing Metabolic Labeled Rare Circulating Tumor Cells Based on Bioorthogonal Chemistry.

Capture of circulating tumor cells (CTCs) with high efficiency and high purity holds great value for potential clinical applications. Besides the existing problems of contamination from blood cells and plasma proteins, unknown/down-regulated expression of targeting markers (e.g., antigen, receptor, etc.) of CTCs have questioned the reliability and general applicability of current CTCs capture methodologies based on immune/aptamer-affinity. Herein, a cell-engineered strategy is designed to break down such barriers by employing the cell metabolism as the leading force to solve key problems. Generally, through an extracellular vesicle generation way, the cell-released magnetic vesicles inherited parent cellular membrane characteristics are produced, and then functionalized with dibenzoazacyclooctyne to target and isolate the metabolic labeled rare CTCs. This strategy offers good reliability and broader possibilities to capture different types of tumor cells, as proven by the capture efficiency above 84% and 82% for A549 and HepG2 cell lines as well as an extremely low detection limitation of 5 cells. Moreover, it enabled high purity enrichment of CTCs from 1mL blood samples of tumor-bearing mice, only ≈5-757 white blood cells are non-specific caught, ignoring the potential phenotypic fluctuation associated with the cancer progression.

A Chimeric Virus-Based Probe Unambiguously Detects Live Circulating Tumor Cells with High Specificity and Sensitivity

The current methods for detecting circulating tumor cells (CTCs) suffer from several drawbacks. We report a novel method that is based on a chimeric virus probe and can detect CTCs with extremely high specificity and sensitivity. Moreover, it exclusively detects live CTCs, and its detection efficacy is not impacted by the variation of EpCAM expression. The chimeric virus probe is composed of a capsid from human papillomavirus that provides the detection with high specificity, and a SV40-based genome that can amplify extensively inside CTCs and, hence, endows the detection n with high sensitivity. Furthermore, different marker genes can be incorporated into the probe to provide detection with versatility. These unique capabilities will likely improve the validity and utility of this CTC detection in several clinical applications, which is one of the drawbacks suffered by many of the current CTC detection methods.

A high-throughput label-free time-stretch acoustofluidic imaging cytometer for single-cell mechanotyping

Current circulating tumor cells (CTC) detection methods have to compromise between sensitivity and throughput. High-throughput imaging cytometer based on serial time-encoded amplified microscopy (STEAM) facilitates CTC detection at single-cell sensitivity from abundant cells. However, this method lacks the information to spot heterogeneity of cells with high morphological similarity. Researches on cell biophysical properties suggest cell mechanotyping can be an indicator of phenotypic heterogeneity to improve classification ability of STEAM cytometer. Here, we present a high-throughput label-free acoustofluidic imaging cytometer for single-cell mechanotyping based on STEAM and acoustofluidic technology. The generated acoustic resonance field translocates cells to different transversal exit positions under continuous flow according to their intrinsic biophysical properties. Such displacements are recorded with images simultaneously using STEAM cytometry at approximately 2000 cells/s. We experimentally verified that our method accounting for both cell images and acoustic displacements can improve mechanotyping accuracy by 12% upon image-based phenotyping method. This new acoustofluidic imaging cytometer facilitates high-accuracy and high-throughput imaging cytometry for single-cell CTC mechanotyping.

Development and Clinical Prospects of Techniques to Separate Circulating Tumor Cells from Peripheral Blood.

Detection of circulating tumor cells (CTC) is an important liquid biopsy technique that has advanced considerably in recent years. To further advance the development of technology for curing cancer, several CTC technologies have been proposed by various research groups. Despite their potential role in early cancer diagnosis and prognosis, CTC methods are currently used for research purposes only, and very few methods have been accepted for clinical applications because of difficulties, including CTC heterogeneity, CTC separation from the blood, and a lack of thorough clinical validation. Although current CTC technologies have not been truly implemented, they possess high potential as future clinical diagnostic techniques for individualized cancer. Here, we review current developments in CTC separation technology. We also explore new CTC detection methods based on telomerase and nanomaterials, such as in vivo flow cytometry. In addition, we discuss the difficulties that must be overcome before CTC can be applied in clinical settings.

Circulating Tumor Cell Detection Technologies and Clinical Utility: Challenges and Opportunities.

The potential clinical utility of circulating tumor cells (CTCs) in the diagnosis and management of cancer has drawn a lot of attention in the past 10 years. CTCs disseminate from tumors into the bloodstream and are believed to carry vital information about tumor onset, progression, and metastasis. In addition, CTCs reflect different biological aspects of the primary tumor they originate from, mainly in their genetic and protein expression. Moreover, emerging evidence indicates that CTC liquid biopsies can be extended beyond prognostication to pharmacodynamic and predictive biomarkers in cancer patient management. A key challenge in harnessing the clinical potential and utility of CTCs is enumerating and isolating these rare heterogeneous cells from a blood sample while allowing downstream CTC analysis. That being said, there have been serious doubts regarding the potential value of CTCs as clinical biomarkers for cancer due to the low number of promising outcomes in the published results. This review aims to present an overview of the current preclinical CTC detection technologies and the advantages and limitations of each sensing platform, while surveying and analyzing the published evidence of the clinical utility of CTCs.

Abstract A089: Detection of circulating tumor cells (CTC) by microfluidic technology based on lateral magnetophoresis and CTC-based multigene profiling analysis in prostate cancer patient

Abstract Introduction By using the circulating tumor cells (CTCs), we can get useful genomic information that enable monitoring of patient’s prognosis or selecting of appropriate treatment over time by repeat biopsy. Despite their usefulness, current CTC separation techniques are limited in terms of their ability to separate high recovery and purity CTCs from peripheral blood. We aimed to separate CTCs from prostate cancer patients using microfluidic technique that based on disposable lateral magnetophoretic microseparator (named ‘assembly-disposable CTC-μChip’) and assess the gene expression that are specific for prostate cancer using mRNA from isolated CTCs measured by droplet digital PCR (ddPCR) method. Patients and Methods Total 54 samples of 45 patients who underwent treatment for prostate cancer between 06/2018 and 03/2019 were prospectively enrolled for CTC collection. Approximately 10 ml of whole blood was drawn from prostate cancer patients; Group A, 12 samples (22.2%) with localized stage II-III cancer, Group B, 5 samples (9.3%) with stage IV, Group C, 6 samples (11.1%) with metastatic hormone-sensitive prostate cancer (mHSPC) and Group D, 31 samples (57.4%) with metastatic castration-resistant prostate cancer (mCRPC). CTCs were positively isolated by using Assembly-disposable CTC-μChip. CTCs from 5 mL of blood were used for counting to identify isolated CTC numbers with immunofluorescence detection procedure. CTCs from the remaining 5 mL of blood were used for gene analysis by ddPCR. For the gene expression analysis, selected 13 patient’s samples were examined for 6 prostate cancer-related genes, such as androgen receptor (AR), androgen receptor splice variant 7 (AR-V7), prostate specific membrane antigen (PSMA), prostate specific antigen (PSA), cytokeratin-19 (CK-19) and epithelial cell adhesion molecule (EpCAM). Results We identified the average of 3, 16, 14 and 26 CTCs per 1ml in group A, B, C and D, respectively (P< 0.01). Background contaminated white blood cells are indispensably isolated with CTCs that counted on average 2245.4 cells, thereby CTC purity rate was 2.56 %. The gene positive rate of AR, AR-V7, PSA, PSMA, KRT-19, and EpCAM were 2.3 % (12/13), 15.4 % (2/13), 38.5 % (5/13), 69.2 % (9/13), and 100 % (13/13), respectively. Especially, the copy number of PSA, PSAM and CK-19 increased in proportion as the clinical stage increased. All samples showed EpCAM positive, whereas AR-V7 was rare events, with only in 2 out 5 patients positive with mCRPC, but not in other stages. Conclusions This study demonstrates that assembly-disposable CTC-μChip is a suitable for CTC isolation from prostate cancer patients. Our data also showed that CTC-based multigene profiling can be analyzed using ddPCR, making it a clinically relevant biomarker. Interrogating tumor expression via CTCs isolation may allow for enhancing precision-based treatment. Citation Format: Hyungseok Cho, Ki-Ho Han, Jae-Seung Chung. Detection of circulating tumor cells (CTC) by microfluidic technology based on lateral magnetophoresis and CTC-based multigene profiling analysis in prostate cancer patient [abstract]. In: Proceedings of the Twelfth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2019 Sep 20-23; San Francisco, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl_2):Abstract nr A089.

Abstract B14: Artificial intelligence assists automation and high performance of circulating tumor cells enumeration and circulating tumor microemboli characterization in fluorescence microscopy images

Abstract Analysis of circulating tumor cells (CTCs) provides exclusive benefits compared with other liquid biopsy techniques due to their advanced clinical applications including single-cell multiomics (DNA, RNA, proteins, and metabolites) analysis and the ability to generate CTCs-derived xenograft models. Current CTC analysis primarily relies on labor-intensive and human-biased manual CTC enumeration or semiautomated work, hindering clinical adaptation of CTC analysis into practice. Beyond CTC enumeration, recent studies focused on detection and characterized features of circulating tumor microemboli (CTM) to improve cancer monitoring and diagnosis, but the added complexity of CTM makes CTC analysis even more challenging. Development of an automated, unbiased, and specialized computation method for CTC analysis remains a clinical unmet need. We developed a deep learning-based framework by applying computer vision to efficiently automate CTC enumeration and characterization with high throughput and accuracy. Fluorescence microscopy CTCs images (CK+/CD45-/DAPI+) have been prepared from blood samples of non-small cell lung cancer patients by the “CMx” CTC capture platform. This study applied an “active learning” model that only requires a small upfront training dataset while being able to accumulate more training data to make improvements with minimal annotation efforts. We have collected 20 images annotated by subject matter experts to train the initial model. Using an iterative process where feedback from the subject matter experts was used to influence the model, we achieved continuous improvements using adaptive annotations. After the model had been well trained and validated by 4 extra images, we used 18 new test images to benchmark the model performance. Preliminary results demonstrated that our AI model outperformed current semiautomated methods having higher sensitivity and reduced time for CTC enumeration in a lung cancer CTC sample analysis. In 18 test samples, the model predicted 34% more total CTCs than the current method (1,775 versus 1,328 counts). By applying the model to the 24 training samples, the model also recovered 45% more total CTCs absent from the original human annotation (2,507 versus 1,732 counts). In some samples, enumeration assisted by the model saved 90% of time over the current method (<20 min vs ~4 hrs.) for CTC enumeration, which would result in an enormous advantage if the datasets being analyzed included a large number of samples. Furthermore, our AI model was able to characterize features of CTM, including CTC clusters and CTC-associated immune cells, in addition to enumerating the samples. The AI model developed in this study is a promising method for CTC analysis that significantly improved throughput, accuracy, and reproducibility, leading to an approach of CTC analysis better suited for clinical adoption. Development of AI-driven CTC/CTM analysis provides additional resources and advancements to the field of liquid biopsy for the diagnosis and treatment of disease. Citation Format: Wei-Lei Yang, Chi-Bin Li, Jia-Yang Chen, Samuel Chen, Ten-Jen Liu, Ying-Chih Chang. Artificial intelligence assists automation and high performance of circulating tumor cells enumeration and circulating tumor microemboli characterization in fluorescence microscopy images [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr B14.