Search Results (129)

Background/Objectives: Breast cancer (BC) is the most commonly diagnosed cancer in women, and metastasis is the leading cause of BC-related death. Circulating tumor cells (CTCs) are a prerequisite for metastasis. This study examined the diagnostic and prognostic value of CTCs for assessing metastatic risk and recurrence in BC. Methods: The Chiline CATCH^® Circulating Target Cell Enrichment System, an automated negative selection platform, was used to enrich and enumerate CTCs from the peripheral blood of patients with BC. Epithelial cell adhesion molecule (EpCAM) and cell-surface Vimentin (CSV) were used as markers for CTC identification. Results: CSV⁺ CTC counts, but not EpCAM⁺ CTC counts, were increased in patients with BC at higher metastatic risk. A cut-off of >4.5 CSV⁺-CTCs/2 mL blood yielded a sensitivity of 0.56 and specificity of 0.92 for identifying patients at high metastatic risk. CSV⁺-CTCs outperformed conventional serum tumor markers, including cancer antigen 15-3 (CA 15-3), cancer antigen 125 (CA 125), and carcinoembryonic antigen (CEA), in identifying patients with high metastatic risk, and their combined use further improved risk stratification. An elevated CSV⁺-CTC count (≥5 cells/2 mL blood) was significantly associated with worse progression-free survival in patients with BC. Conclusions: These findings suggest that CSV⁺-CTCs may serve as a biomarker for metastatic risk stratification and recurrence monitoring in BC when measured using an automated negative selection platform. Full article

Circulating tumor cells (CTCs) are valuable liquid-biopsy biomarkers, yet their extreme rarity makes high-purity, high-throughput enrichment challenging. In spiral inertial microfluidics, high cell loading induces long-range hydrodynamic interactions that broaden the focused blood-cell stream; consequently, a subpopulation completes the ~0.5 and ~1.0 Dean-cycle migrations with a phase delay, compressing the CTC–blood cell gap and degrading purity. Here we propose a Dean-cycle phase-regulated double-spiral design informed by this phenomenon. This design aims to mitigate the stream-broadening effect by boosting the Dean number during the first half-cycle to promote synchronized blood-cell migration and shifting the CTC equilibrium position near one full cycle to further widen the CTC–blood cell separation. We implement this strategy in a second-generation double-spiral microfluidic chip (SDMC) and scale it to a four-channel parallel array (ASDMC). Under optimized conditions, ASDMC processes diluted whole blood (hematocrit = 4%) without the need for red blood cell (RBC) lysis or antibody labeling, achieving a sample throughput of 1200 μL·min⁻¹. Specifically, it exhibits a mean recovery rate of 98.8% across three spiked tumor cell lines (MCF-7, PC-9, and Mahlavu) and a mean white blood cell (WBC) depletion efficiency of 93.3%. In a pilot clinical testing of 20 patients (NSCLC and HCC), enriched fractions enabled immunofluorescence identification of CK⁺CD45⁻DAPI⁺ CTCs, with an exploratory trend of increasing CTC counts with advanced disease stage (4–34 cells·mL⁻¹). These results describe a scalable, label-free platform, and the observed purification performance aligns with our proposed mechanism: Dean-cycle phase regulation to mitigate blood-cell migration lag. Our findings support further technical validation and clinical assessment in larger cohorts. Full article

With the growing demand for personalized design, residential layout generation has become a key research area in architecture and artificial intelligence. This paper presents a novel method for generating personalized layouts using an improved Conditional Variational Autoencoder (HCVEA). This method introduces conditional variables to control key features such as room count, functional zoning, and spatial distribution, while enhancing the latent space structure to improve both diversity and controllability. By incorporating user preferences as conditional inputs and integrating reinforcement learning with the autoencoder-based architecture, the layout generation process is optimized for more accurate and efficient results. A novel Connectionist Temporal Classification Attention (CTC-Attention) decoder is introduced to improve contextual semantic understanding. The Asynchronous Advantage Actor–Critic (A3C) reinforcement learning algorithm is employed to enhance training efficiency. Experimental results averaged over three independent runs show that HCVEA achieves an FZMR of 90.7% and an RAR of 92.3% with low standard deviation, outperforming baseline models. It also maintains a constraint compliance rate of 88.6% and adaptability to different room configurations. Full article

Circulating tumor cells (CTCs) are shed from the primary tumor into the bloodstream and represent dynamic molecular biomarkers for monitoring the progression of cancer. While profiling tumor tissues with over expression of cell surface markers, such as PD-L1 or HER2, is standard in guiding therapy, tissue samples are often inaccessible and inadequate, especially post-surgery or in cases of recurrence. Emerging clinical evidence indicates that CTC counts and biomarker surface expression can predict prognosis and therapeutic resistance more accurately than imaging or tissue-based approaches. Recent advancements in the CTC detection methods, based on physical properties or surface markers (e.g., EpCAM), coupled with next-generation sequencing (NGS) have enabled the isolation of these rare cells and their molecular characterization. Consequently, CTCs provide a real-time alternative, enabling repeated, longitudinal assessment of tumor phenotype and therapeutic response. This review emphasizes the translational potential of surface protein biomarkers on CTCs for profiling, namely PD-L1, HER2, and EGFR, as a clinically actionable approach to stratify patients, guide immunotherapy decisions, and monitor minimal residual disease (MRD), especially when longitudinal tissue biopsies are not feasible. Full article

Lung cancer is the leading cause of cancer-related mortality worldwide, with non-small-cell lung cancer (NSCLC) accounting for the majority of cases. Standard tissue biopsies are invasive and unsuitable for repeated monitoring. Liquid biopsy technologies, particularly circulating tumor cell (CTC) analysis, offer a minimally invasive alternative for real-time disease tracking. To address the need for efficient and reproducible CTC isolation, we developed the Cellsway microfluidic CTC enrichment and identification platform, which employs inertial hydrodynamics in a spiral-shaped microfluidic channel comprising hydrofoil-shaped pillars to enable high-throughput, label-free enrichment of CTCs while preserving cell integrity, followed by an optimized CTC identification assay. Analytical performance assessed through spiking experiments using NSCLC cell lines demonstrated recovery rates of 91.9% for H1975 cells and 78.3% for A549 cells. Clinical validation was performed on blood samples from 51 stage IV NSCLC patients. A 7.5 mL volume of peripheral blood was processed with the SwayBox platform, and enriched CTCs were identified through an optimized multiplex immunofluorescence protocol. CTCs were detected in 47% of NSCLC patients, with counts ranging from 0 to 72 cells per 7.5 mL of blood. At a cutoff of 1 CTC per 7.5 mL, the assay achieved a specificity of 95%. Patient-derived CTCs exhibited smaller mean diameters compared to cultured NSCLC cell lines, yet were effectively enriched through hydro-dynamic tuning. These findings demonstrate that the Cellsway platform enables efficient and re-producible CTC isolation with high specificity, supporting its potential utility for clinical monitoring and precision oncology in NSCLC. Full article

Lung cancer, as one of the most prevalent and lethal malignancies, requires immediate and effective therapeutic solutions. Therefore, additional innovative methods are continually sought to achieve optimal treatment outcomes. Various markers are used to select the most effective therapies, assess clinical responses, and facilitate follow-up care for the patients. Circulating tumor cells (CTCs) remain a valuable biomarker in clinical management of cancer patients due to the range of information they provide and their high prognostic and predictive potential in monitoring anticancer therapy. CTCs constitute a heterogeneous population of cancer cells that undergo an epithelial-to-mesenchymal transition (EMT), are shed from the tumor mass, and migrate through the peripheral blood, ultimately causing metastases. In this literature review, we focus on the biological, biochemical, and biophysical properties of CTCs, specifically from the perspective of the design of CTC enumeration technologies. Furthermore, we combine the available data on the application of CTC count in monitoring various treatment modalities in lung cancer, including radiotherapy, chemotherapy, tyrosine kinase inhibitors, and immunotherapy. Although many published reports indicate that an increased number of CTCs in blood samples of lung cancer patients correlates with worse treatment outcomes, several limitations hinder the widespread usage of CTCs in the clinical setting. Full article

Background: The detection of circulating tumor cells (CTCs) holds significant promise for the diagnosis and monitoring of lung cancer (LC). However, the clinical utility of CTCs is limited by the heterogeneity of their phenotypes and the shortcomings of existing detection methods, which often rely on epithelial markers like EpCAM. DNA aptamers offer a promising alternative due to their high affinity, stability, and ability to recognize diverse cancer-specific biomarkers. Methods: This study utilized DNA aptamers LC-17 and LC-18, previously selected against primary lung tumor tissue, to isolate and detect CTCs in the peripheral blood of 43 non-small cell lung cancer (NSCLC) patients. Mass spectrometry (LC-MS/MS) was employed to identify the target proteins of aptamer LC-17. CTCs from patients’ blood and healthy donors were isolated via filtration after erythrocyte and lymphocyte lysis and stained with FAM-labeled LC-17 and LC-18 aptamers for detection using fluorescence and light microscopy. Results: Mass spectrometry identified neutrophil defensin 1 (DEFA1) and peroxiredoxin-2 (PRDX2) as the primary protein targets of aptamer LC-17 in CTCs, both of which were absent in healthy donor samples. CTC enumeration revealed statistically significant correlations between elevated CTC counts (>3 cells/4 mL blood) and advanced primary tumor size (T4 vs. T1–T3, p = 0.012), extensive regional lymph node metastasis (N3 vs. N1–N2, p = 0.014), and shorter overall survival (median 24 vs. 32 months, p < 0.05). Conclusions: The developed aptamer-based liquid biopsy method effectively captures heterogeneous CTC populations independent of EpCAM expression. The strong correlation of CTC counts with disease progression and survival underscores their clinical relevance as a prognostic biomarker in NSCLC. This approach presents a viable, non-invasive tool for disease monitoring and stratification of NSCLC patients, with potential for integration into clinical practice. Full article

Noninvasive liquid biopsy for monitoring circulating tumor cells offers valuable insights for predicting therapeutic responses. We developed TelomeScan^® (OBP-401), based on the detection of telomerase activity as a universal cancer cell marker and an indicator of the presence of viable circulating tumor cells (CTCs) for patients with advanced non-small cell lung cancer (NSCLC). This system evaluated CTC subtypes characterized by programmed death ligand 1 (PD-L1), an immune checkpoint molecule, and vimentin, an epithelial–mesenchymal transition (EMT) marker, using a multi-fluorescent color microscope reader. The prognostic value and therapeutic responses were predicted by dynamically monitoring CTC counts in 79 patients with advanced NSCLC. The sensitivity and specificity values of TelomeScan^® for PD-L1⁽⁺⁾ cells (≥1 cell) were 75% and 100%, respectively, indicating high diagnostic accuracy. PD-L1⁽⁺⁾ and EMT⁽⁺⁾ in CTCs were detected in 75% and 12% of patients, respectively. Detection of PD-L1⁽⁺⁾CTCs and PD-L1⁽⁺⁾EMT⁽⁺⁾ CTCs before treatment was associated with poor prognosis (p < 0.05). Monitoring of reducing and increasing PD-L1⁽⁺⁾ CTC counts in two sequential samples (baseline, cycle 2 treatment) correlated significantly with partial response (p = 0.032) and progressive disease (p = 0.023), respectively. Monitoring PD-L1⁽⁺⁾CTCs by TelomeScan^® will aid in anticipating responses or resistance to frontline treatments, optimizing precision medicine choices in patients with NSCLC. Full article

Circulating tumor cells (CTCs) are multifaceted biomarkers with significant potential for precision oncology, offering opportunities to refine diagnoses and personalize treatments across various cancer types, including colorectal and breast cancer. CTC assays serve as reliable prognostic indicators, even during chemotherapy and/or molecularly targeted therapies. Notably, CTCs exhibit heterogeneity that gradually develops during carcinogenesis and becomes more pronounced in advanced disease stages. These intra- and intertumoral heterogeneities pose challenges, particularly when drug-resistant clones emerge following therapy. The dynamic behavior of CTCs provides valuable insights into treatment response and prognosis. Extensive efforts have led to the development of technologies for effective CTC isolation, accelerating their clinical implementation. While both CTC and circulating tumor DNA (ctDNA) tests offer prognostic value, they reflect different aspects of tumor biology: CTC counts indicate tumor progression, while ctDNA levels correlate with tumor burden. The combined analysis is expected to yield complementary insights. CTC tests are feasible in general hospitals and may serve as tumor markers comparable to, or even superior to, conventional markers such as carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) for colorectal cancer, and CA15-3 for breast cancer. Early incorporation of CTC tests into routine blood panels appears to be a rational and promising approach. Full article

Investigating the molecular and genetic characteristics of circulating tumor cells (CTCs) presents a promising approach for personalizing treatment in patients with malignant neoplasms, given the limitations of traditional biopsy and histopathology. This study aimed to isolate, characterize, and analyze CTC dynamics in the peripheral blood of 30 patients with metastatic lung cancer to develop criteria for treatment response and prognosis. We detected CTCs before the start of the treatment and monitored changes during treatment, correlating these with responses evaluated by standard imaging methods. A decrease in the CTCs in the course of the therapy was linked to a favorable tumor response, while the stable CTC counts indicated a lack of response and poor survival prognosis. The OS of patients was analyzed and compared with the initial number of CTCs in peripheral blood samples. The significant reductions in median OS were evident in patients with >3 total CTCs at baseline compared to those with ≤3 total CTCs (median survival 26 months, n = 10, vs. median survival 8 months, n = 19, respectively with HR = 2.6, 95% CI 1.07 to 6.4). Full article

Background: Circulating tumor cells (CTCs) have recently been developed as biomarkers. Several studies have reported on the clinical use of CTCs to assess drug resistance in various cancers. However, sequential and multiple CTC measurements during chemotherapy are relatively rare. We recently reported a transient increase in CTCs early after chemotherapy by sequentially detecting CTCs in a human pancreatic cancer xenograft model in nude mice. Method: In the present study, using a newly developed immunocytology and glass slide-based convenient CTC detection platform, we examined CTC numbers sequentially before, during, and after chemotherapy in the peripheral blood of 14 pancreatic cancer patients, pathological stage (pStage) I-IV, who underwent surgery with preoperative chemotherapy and GS (Gem/S-1) and GnP (Gem/nab-PTX). Results: Among patients with strongly or weakly elevated CTC counts (3–44/5 mL of blood) following GS treatment, four out of six pancreatic cancer patients were judged to have a partial response (PR), and two out of six were deemed to have stable disease (SD) as a clinical response based on the CT image. In contrast, in patients with GnP therapy, three out of four patients showed no CTC response, and these three patients were judged to have progressive disease (PD), while the remaining one patient was judged to have SD in terms of their clinical response. Conclusion: These results suggest that sequential CTC monitoring during preoperative chemotherapy in pancreatic cancer patients can be a helpful liquid biopsy diagnostic tool as a therapeutic marker to predict tumor chemosensitivity and chemoresistance in clinical settings. Further large-scale clinical studies are required to confirm and clarify this hypothesis. Full article

Prostate cancer is a leading cause of cancer-related mortality in men, with radiotherapy (RT) playing a pivotal role in treatment. However, reliable biomarkers for assessing relapse risk following RT remain scarce. This study aimed to evaluate circulating epithelial tumor cells (CETC/CTC) as potential biomarkers for assessing relapse risk in prostate cancer patients undergoing RT. Peripheral blood samples were collected from 52 prostate cancer patients, and CETC/CTC were detected using the EpCAM surface marker. Patients received definitive, adjuvant, or salvage RT, and CETC/CTC counts were measured before, at mid-treatment, and at the end of RT. The association between changes in CETC/CTC counts and relapse risk was examined. CETC/CTC were detected in 96% of patients prior to RT. A significant reduction in CETC/CTC counts during RT, particularly in patients who had undergone surgery, was associated with a lower relapse risk. In contrast, an increase in CETC/CTC counts during or after RT was associated with a higher relapse risk (hazard ratio = 8.8; p = 0.002). Furthermore, 36% of patients receiving adjuvant RT and 14% of those receiving definitive RT relapsed, with a higher risk observed in patients showing increasing CETC/CTC counts during RT. Among patients receiving salvage RT, 18% relapsed, though changes in CETC/CTC counts were less significantly associated with relapse. Monitoring CETC/CTC levels during RT offers important prognostic insights into relapse risk in prostate cancer patients. While changes in CETC/CTC counts correlated with relapse, PSA levels measured during the study did not reliably reflect relapse risk in this cohort. CETC/CTC shows promise as a prognostic marker, though further studies are required to validate its clinical superiority over PSA. Full article

Hepatocellular carcinoma (HCC) is the sixth most common cancer and the third leading cause of cancer-related death worldwide, with no precise method for early detection. Circulating tumor cells (CTCs) expressing the dynamic polarity of the cytoskeletal membrane protein, ezrin, have been proposed to play a crucial role in tumor progression and metastasis. This study investigated the diagnostic and prognostic potential of polarized circulating tumor cells (p-CTCs) in HCC patients. CTCs were isolated from the peripheral blood of 20 HCC patients and 18 patients with nonmalignant liver disease (NMLD) via an OncoQuick^® kit and immunostained with Ezrin-Alexa Fluor 488^®, CD146-PE, and CD45-APC. A fluorescence microscopy was then performed for analysis. The HCC group exhibited significantly higher levels of p-CTCs, with median values of 0.56 p-CTCs/mL, compared to 0.02 p-CTCs/mL (p = 0.03) in the NMLD group. CTCs were detected in 95% of the HCC patients, with a sensitivity of 95% and specificity of 89%. p-CTCs were present in 75% of the HCC patients, with a sensitivity of 75% and a specificity of 94%. Higher p-CTC counts were associated with the significantly longer overall survival in HCC patients (p = 0.05). These findings suggest that p-CTCs could serve as valuable diagnostic and prognostic markers for HCC. The incorporation of p-CTCs into diagnostic strategies could enhance therapeutic decision-making and improve patient outcomes. Full article

Background: Relapsed unresectable triple-negative breast cancer is a demanding disease with only a few treatment options. Especially for patients with unresectable tumor masses, a treatment that offers rapid tumor shrinkage is needed. If patients are exhausted from several treatment lines, systemic side effects have to be avoided. Reversible electroporation has shown to be effective for breast cancer if combined with systemic bleomycin and/or cisplatin. To enhance the local effect and reduce the systemic side effects, we combined reversible electroporation with regional chemotherapy. Materials and Methods: Patients with advanced metastasized and relapsed breast cancer received regional chemotherapy via intra-arterial infusion and isolated thoracic perfusion combined with percutanous reversible electroporation. Circulating tumor cells (CETCs/CTCs) were counted before and 24 h after the treatment. Tumor response was evaluated by CT (computer tomography) control. Results: A total of 21 treatments were conducted for 14 patients who had a mean tumor size of 7.6 cm (standard deviation 3.3 cm). Higher local drug levels are present with arterial infusion compared to venous infusion and result in enhanced response rates. Circulating tumor cells decreased or stayed stable for 24 h after the treatment for 11 and 8 cases, respectively. An increase was observed in two cases. A total of 13 patients showed a clinical response with tumor shrinkage that led to resectability. One patient did not respond to the treatment regimen. Conclusions: The combination of reversible electroporation with intra-arterial chemotherapy is feasible and results in a good clinical response with neglectable side effects. The treatment is repeatable and can lead to resectability. Full article

Stress-inducible heat shock protein 70 (Hsp70), which functions as a molecular chaperone and is frequently overexpressed in different cancer cell types, is present on the cell surface of tumor cells and is actively released into the circulation in free and extracellular lipid vesicle-associated forms. Since the exact pathomechanism of endometriosis has not yet been elucidated (although it has been associated with the development of endometrial and ovarian cancer), we asked whether extracellular Hsp70 and circulating endometriotic cells (CECs) reflect the presence and development of endometriosis. Therefore, circulating levels of free and lipid microvesicle-associated Hsp70 were measured using the Hsp70-exo ELISA, and the presence of circulating CECs in the peripheral blood of patients with endometriosis was determined using membrane Hsp70 (mHsp70) and EpCAM monoclonal antibody (mAb)-based bead isolation approaches. Isolated CECs were further characterized by immunofluorescence using reagents directed against cytokeratin (epithelial marker), CD45 (leukocyte marker), CD105/CD44 (mesenchymal stemness markers) and by comparative RNA analysis. Similar to the situation in patients with cancer, the levels of circulating Hsp70 were elevated in the blood of patients with histologically proven endometriosis compared to a healthy control cohort, with significantly elevated Hsp70 levels in endometriosis patients with lesions outside the uterine cavity. Moreover, CECs could be isolated using the cmHsp70.1 mAb-based, and to a lesser extent EpCAM mAb-based, bead approach in all patients with endometriosis, with the highest counts obtained using the mHsp70-targeting procedure in patients with extra-uterine involvement. The longevity in cell culture and the expression of the cytokeratins CD105 and CD44, together with differentially expressed genes related to epithelial-to-mesenchymal transition (EMT), revealed similarities between mHsp70-expressing CECs and circulating tumor cells (CTCs) and suggest a mesenchymal stem cell origin. These findings support the involvement of mHsp70-positive stem cell-like cells in the development of endometriotic lesions. In summary, elevated levels of Hsp70 and CECs in the circulation could serve as liquid biopsy markers for endometriosis with extra-uterine involvement and help to elucidate the underlying pathomechanism of the disease. Full article