Detection Of Tumor Cells Research Articles

Microfluidic platform for circulating tumor cells isolation and detection

Microfluidic platform for circulating tumor cells isolation and detection

A Systematic Mapping Study of detection of Tumor Cell Targeted by Enzymes though Cerebrospinal Fluid

Cancers, especially of the neural tissue, are often deemed a death sentence. There is, however, still no clear understanding of the underlying causes nor the cellular and molecular mechanisms of this disease. The spread and collateral damage caused by various types of brain tumors remain poorly understood, despite the information that is currently available about these diseases. A common means of diagnosing tumors is through cerebrospinal fluid (CSF) leakage, and the enzymes from brain cancer ca

Activated Leucocyte Cell Adhesion Molecule (CD166): A Biomarker in Diagnosis and Prognosis of Breast Cancer

The global incidence of breast cancer and associated mortality is rising annually despite advanced diagnostic aids and treatment methods. This is due to the failure and difficulty in detecting the disease at an early stage. Due to an increase in the breast cancer mortality rate, biomarkers associated with early and prompt detection are being extensively studied, owing to their sensitivity, specificity, cost-effectiveness, and minimally invasive or non invasive analysis. Among those carbohydrate antigens, carcinoembryonic antigens, and circulating tumour cells are the most commonly observed serum biomarkers in the detection of cancer. In recent years, extensive research is carried out on cell adhesion molecules and their role in the process of cell adhesion which is essential for the development and homeostasis in multicellular organisms. These cell adhesion molecules accelerate appropriate cell response and intercellular communication. Activated Leucocyte Cell Adhesion Molecule (ALCAM) or Cluster of Differentiation 166 (CD166) is a transmembrane glycoprotein and emerging as a promising biomarker for primary detection of tumour cells with metastatic potential and is also upregulated in breast tumours. This review ascertains the usefulness of the ALCAM as a potential diagnostic and prognostic marker for early detection, classification, and prognosis of breast cancer and also elucidates the role of the various other biomarkers.

The Value of Circulating Tumor Cells and Tumor Markers Detection in Lung Cancer Diagnosis.

Circulating tumor cells are complete tumor cells with multi-scale analysis values that present a high potential for lung cancer diagnosis. To enhance the accuracy of lung cancer diagnosis, we detected circulating tumor cells by the innovated conical micro filter integrated microfluidic system. We recruited 45 subjects of study, including 22 lung cancer patients, 2 precancerous patients, the control group including 14 healthy participants, and 7 patients with lung benign lesions in this prospective study. We calculated the area under the receiver operating characteristic curve of circulating tumor cells, cytokeratin19 fragment, carcinoma embryonic antigen, squamous cell carcinoma, neuron-specific enolase, and their combination, respectively, while compared the circulating tumor cells levels between vein blood and arterial blood. A conical shape filter embedded in a microfluidic chip was used to improve the detection capability of circulating tumor cells. The study indicated that the sensitivity, specificity, positive predictive value, and negative predictive value of circulating tumor cells detection were 81.8%, 90.5%, 90.0%, and 82.6%, respectively. The circulating tumor cells level of lung cancer patient was significantly higher than that of the control group (P < .05). The area under the curve of circulating tumor cells, cytokeratin19 fragment, carcinoma embryonic antigen, squamous cell carcinoma, and neuron-specific enolase alone was 0.838, 0.760, 0.705, 0.614, and 0.636, respectively. The combination area under the curve of the 4 tumor markers (cytokeratin19 fragment, carcinoma embryonic antigen, squamous cell carcinoma, and neuron-specific enolase) was 0.805 less than that of circulating tumor cells alone. Together, the total area under the curve of circulating tumor cell and the 4 tumor markers were 0.847, showing the highest area under the curve value among all biomarkers. In addition, this study found that there was no significant difference in positive rate of circulating tumor cell between arterial and venous blood samples. The circulating tumor cells detection technology by conical micro filter integrated microfluidic could be used for lung cancer diagnosis with high sensitivity and specificity. Complementary combination of circulating tumor cells and conventional 4 lung cancer markers could enhance the clinical application accuracy. Venous blood should be used as a routine sample for circulating tumor cells detections.

Enrichment of Circulating Tumor Cells of Lung Cancer and Correlation With Serum Leukomonocyte and Tumor Biomarkers: A Retrospective Study.

Circulating tumor cells is important in the clinical diagnosis of cancer and there are a number of circulating tumor cell detection systems associated with different isolation strategies being validated. There is a novel platform, the CytoBot 2000, which utilizes a combination of physical and immunological technologies to isolate and capture circulating tumor cells. In this retrospective study, 39 lung cancer patients and 11 normal healthy individuals were enrolled and performed circulating tumor cell tests and immunofluorescence staining with CytoBot 2000. The performance of this device was assessed by receiver operating characteristic curve. The clinical relevance of circulating tumor cells was assessed by Chi-square. The correlations between circulating tumor cell number and blood lymphocytes and tumor biomarkers were analyzed by Pearson correlation coefficient. The number of circulating tumor cell is significantly increased in lung cancer patients (3.74 > 0.45, P < .0001). The CytoBot 2000 presented a 100% (39/39) circulating tumor cell detection rate in lung cancer patients and 36% (4/11) in healthy individual blood samples, the sensitivity and specificity were 89.7% and 90.9%, respectively, and with the area under curve of 0.966. Further, there was a positive correlation between circulating tumor cell count and carcinoembryonic antigen 211 (R2 = 0.125, P = .027), but not blood lymphocytes (P = .089). This automatic platform showed excellent performance of circulating tumor cell detection by clinical sample. The tumor biomarkers increased with the number of circulating tumor cell in the lung cancer patients.

Inflammatory pseudotumor-like follicular dendritic cell sarcoma with first clinical manifestation of thrombocytopenia: A case report.

Inflammatory pseudotumor-like follicular dendritic cell sarcoma (IPT-like FDCS) is often associated with Epstein-Barr (EB) virus infection. The tumor is commonly found in the spleen and liver, and it has been reported in the literature that it can be associated with paraneoplastic pemphigus, myasthenia gravis, and other diseases. A case of IPT-like FDCS with clinical features of thrombocytopenia has not been reported. A 59-year-old male patient visited our hospital in September 2020 due to bleeding gums and epistaxis. Splenic lymphoma with marked thrombocytopenia was initially diagnosed. The patient underwent pathological examination after splenectomy. Microscopic examination showed spindle-shaped or oval cells arranged in loose bundles, a large number of lymphocytes and plasma cells infiltrating the interstitium, and fibrin-like changes in the blood vessel wall. Immunohistochemical detection of tumor cells was positive for CD21, CD35, and Epstein-Barr virus in situ hybridization, and the patient was diagnosed with IPT-like FDCS. The patient underwent a splenectomy. The patient received platelet-raising therapy postoperatively. No tumor recurrence or metastasis was found during the 17-month follow-up period, and the platelet count returned to normal. IPT-like FDCS is an uncommon tumor, and its first presentation with marked thrombocytopenia is even rarer. The tumor was clinically and radiographically nonspecific. Definitive diagnosis relies on histopathological and immunohistochemical staining. IPT-like FDCS is biologically indolent and has a favorable prognosis.

Early Detection of Leptomeningeal Metastases Among Patients Undergoing Spinal Stereotactic Radiosurgery

PurposeThe management of patients with advanced solid malignancies increasingly uses stereotactic body radiation therapy (SBRT). Advanced cancer patients are at risk for developing leptomeningeal metastasis (LM), a fatal complication of metastatic cancer. Cerebrospinal fluid (CSF) is routinely collected during computed tomography (CT) myelography for spinal SBRT planning, offering an opportunity for early LM detection by CSF cytology in the absence of radiographic LM or LM symptoms (subclinical LM). This study tested the hypothesis that early detection of tumor cells in CSF in patients undergoing spine SBRT portends a similarly poor prognosis compared with clinically overt LM. Methods and MaterialsWe retrospectively analyzed clinical records for 495 patients with metastatic solid tumors who underwent CT myelography for spinal SBRT planning at a single institution from 2014 to 2019. ResultsAmong patients planned for SBRT, 51 (10.3%) developed LM. Eight patients (1.6%) had subclinical LM. Median survival with LM was similar between patients with subclinical versus clinically evident LM (3.6 vs 3.0 months, P = .30). Patients harboring both parenchymal brain metastases and LM (29/51) demonstrated shorter survival than those with LM alone (2.4 vs 7.1 months, P = .02). ConclusionsLM remains a fatal complication of metastatic cancer. Subclinical LM detected by CSF cytology in spine SBRT patients has a similarly poor prognosis compared with standardly detected LM and warrants consideration of central nervous system-directed therapies. As aggressive local therapies are increasingly used for metastatic patients, more sensitive CSF evaluation may further identify patients with subclinical LM and should be evaluated prospectively.

Sensitive detection of folate receptor-positive circulating tumor cells based on intracellular uptake of the PbS nanoparticle cluster-loaded phospholipid micelles decorated with folic acid in combination with E-DNA sensor

Developing methods for the circulating tumor cells (CTCs) detection is important for noninvasive cancer diagnosis. A sensitive method for homogeneous electrochemical detection of CTCs was developed based on the affinity of folic acid (FA) for folate receptor (FR), which is overexpressed on the CTC surface. The PbS nanoparticle cluster-loaded phospholipid micelles decorated with FA were prepared and used as a nanoprobe. The specific binding of FA to FR-positive CTCs leads to the uptake of the nanoprobes into the CTCs. Cellular imaging assay showed the excellent biocompatibility and selectivity of nanoprobes. After separation of intracellular nanoprobes and the residual free nanoprobes in the solution by centrifugation, nanoprobes in the cell were dissolved using acid. The amount of the target cells was quantified by the electrochemical detection of Pd2+ using the electrochemical DNA (E-DNA) sensor, which was fabricated based on the self-assembly of probe DNA on the flowerlike gold nanostructures modified the screen-printed carbon electrode (SPCE). Because of the large quantities of detectable Pd2+ contained in the nanoprobes, the signal amplification effect was achieved. The developed electrochemical assay allows the detection of ∼5 cells from 1 mL of sample. Studies with both healthy individuals and cancer patient blood samples demonstrate that the assay is sensitive and FR-positive CTCs specific.

Robot based Transurethral Bladder Tumor Resection with automatic detection of tumor cells

Thehandiness, perception, and risks of bladder wall perforation with Transurethral Bladder Tumor Resection (TBRD) have been challenging to process and diagnose due to muscle-invasive bladder tumors. Survey shows in the medical sector, TBTR is usually collected via a rigid resectoscope. Further, the deficiencies in mobility are increased when tumors in the bladder neck area are established. A TBTR Endoscopic Robotic System (ERS) is designed to overcome these limitations. This paper describes the challenges of designing, modeling, and controlling the first TBTR in the robot framework. A Master-Slave robot consists of a continuous cross-sectional robot that can control the ablation, the grasp, and fiber optics with a micro-snake-continuous robot. The strategy for limited telemanipulation, based on redundancy, is consistent with the degree of limitation where the robot experiences are transurethral. The Deep Learning assisted Multi-Scale Feature Fusion Algorithm (MSFFA) has been used to track hundreds of cells with a fully automated and efficient system. Finally, a retracing algorithm combines historical data from matching objects. The results are promising and show that the proposed approach is consistent with the other established methods, which means that potential applications are likely to exist in biomedical engineering.

Identification of Cancer Cells in the Human Body by Anti-Telomerase Peptide Antibody: Towards the Isolation of Circulating Tumor Cells.

Early detection of tumor cells by identifying universal Tumor Associated Antigens (TAA) can drastically change our diagnostic, theranostic and therapeutic possibilities to cure cancer. Human Telomerase Reverse Transcriptase (hTERT), a hallmark of cancer, could act as an optimal TAA candidate. Here we report about the development of a monoclonal antibody against hTERT peptide (α-hTERT mAb) presented on the surface of cancer cells and its possible applications as a pan-cancer marker. Liquid biopsies, an innovative tool in precision oncology, comprising the noninvasive analysis of circulating tumor-derived material to counteract limitations associated with tissue biopsies. Within the tumor circulome, the US Food and Drug Administration already approved the use of circulating tumor cells (CTCs) as valid liquid biopsies. However, currently CTCs are being trapped using antibodies against specific cancer types, with anti EpCAM as the most common antibody, directed mainly against solid tumors. Moreover, the precision medicine approach is based on specific cancer type directed antibodies. Our novel mAb against the hTERT 16-mer peptide, corresponding to amino acids 611-626, is capable of detecting various types of cancer cells both in vitro and ex vivo from tumors of patients with either hematological or solid tumors. This antibody does not bind to normal lymphocytes cells. Cleavage of our antibody to F(ab')2 fragments increased its binding specificity to the tested cancer cells. Future studies may point to the use of this antibody in the procedure of capturing CTCs.

Single-cell sequencing technologies in bladder cancer research: Applications and challenges.

Bladder cancer is among the most common malignant tumors with highly heterogeneous molecular characteristics. Despite advancements of the available therapeutic options, several bladder cancer patients exhibit unsatisfactory clinical outcomes. The lack of specific biomarkers for effective targeted therapy or immunotherapy remains a major obstacle in treating bladder cancer. The rapid development of single-cell techniques is transforming our understanding of the intra-tumoral heterogeneity, thereby providing us with a powerful high-throughput sequencing tool that can reveal tumorigenesis, progression, and invasion in bladder tumors. In this review, we summarise and discuss how single-cell sequencing technologies have been applied in bladder cancer research, to advance our collective knowledge on the heterogeneity of bladder tumor cells, as well as to provide new insights into the complex ecosystem of the tumor microenvironment. The application of single-cell approaches also uncovers the therapeutic resistance mechanism in bladder cancer and facilitates the detection of urinary-exfoliated tumor cells. Moreover, benefiting from the powerful technical advantages of single-cell techniques, several key therapeutic targets and prognostic models of bladder cancer have been identified. It is hoped that this paper can provide novel insights into the precision medicine of bladder cancer.

Hierarchal polyaniline-folic acid nanostructures act as a platform for electrochemical detection of tumor cells

The fabrication of electrochemical sensing platforms for cancer monitoring by quantifying circulating tumor cells (CTCs) in blood holds promise for providing a low-cost, rapid, feasible, and safe approach for cancer diagnosis. Here, we isolate cancer cells using CoFe2O4 nanoparticles functionalized with folic acid and chitosan as an inexpensive magnetic nanoprobe. This electrochemical cytosensing platform was realized using polyaniline-folic acid nanohybrids with a three-dimensional hierarchical structure that presents abundant affinity sites toward overexpressed folate bioreceptors on cancer cells, in addition to retaining satisfied conductivity. Furthermore, 3D modeling and simulation of the polyaniline-folic acid structures were conducted to investigate the stable complex between aniline and folate, and the interaction between the polyaniline-folate complex and folate receptor alpha1, a bioreceptor on MCF-7 was revealed for the first time. The limit of detection was calculated to be 4 cells mL−1 with a linear range from 50 to 106 cells mL−1.

Synthesis of a Dual-Color Fluorescent Dendrimer for Diagnosis of Cancer Metastasis in Lymph Nodes.

Detection of cancer metastasis spread in lymph nodes is important in cancer diagnosis. In this study, a fluorescence imaging probe was designed for the detection of both lymph node and tumor cells using always-ON and activatable fluorescence probes with different colors. Rhodamine B (Rho), a matrix metalloproteinase−2 (MMP−2)-responsive green fluorescence probe, and a tumor-homing peptide were conjugated to a carboxy-terminal dendrimer that readily accumulates in lymph nodes. The activatable green fluorescence signal increased in the presence of MMP−2, which is secreted by tumor cells. Both the always-ON Rho signal and the activatable green fluorescence signal were observed from tumor cells, but only the weak always-ON Rho signal was from immune cells. Thus, this type of dendrimer may be useful for non-invasive imaging to diagnose cancer metastasis in lymph nodes.

Label-free tumor cell screening based on IDO1-mediated tryptophan metabolism at single cell level

Indoleamine 2,3-dioxygenase 1 (IDO1) plays a critical role in inflammatory and immunometabolism programming through catalyzing the oxidation of tryptophan (Trp) into downstream N-formylkynurenine. IDO1 is typically up-regulated in malignant tumors, making it a potential biomarker for cancer diagnosis. Here we show an effective strategy for tumor cell detection by integrating IDO1 activity assay with single cell-encapsulated droplets on a microfluidic platform for high-throughput bioanalysis. Mixed cells, as well as other cofactors, are encapsulated in individual droplets, which act as dynamic microreactors for IDO1-catalyzed oxidation of Trp. After pico-injection of a biosensing ensemble consisting of the macrocycle cucurbit [8]uril (Q8) and a fluorescent guest, rapid and robust screening of tumor cells by fluorescence signal is achieved in a few minutes reporting to Trp depletion, expanding the scope of conventional antibody-based detection of protein biomarkers. The results represent the first example of quantifying IDO1 enzymatic activity at the single cell level with a high-throughput performance, therefore promising warning signs and early diagnosis of tumor cells.

Low-Cost Antenna-Array-Based Metamaterials for Non-Invasive Early-Stage Breast Tumor Detection in the Human Body.

Microstrip patch antennas have been used in many applications since their appearance. Despite their great promise, their narrow bandwidth and the loss at high-frequency bands have limited their usage in medical applications. This work proposes a developed low-cost microstrip patch antenna suitable for microwave imaging (MWI) applications within the wideband frequency range. The proposed antenna is loaded with an artificial magnetic conductor (AMC) to improve the antenna performance. The simulated results obtained using computer simulation technology (CST) indicate that the presence of the AMC has improved the frequency selectivity of the antenna at 8.6 GHz with a peak realized gain of 9.90443 dBi and 10.61 dBi for simulated and measured results, respectively. The proposed microstrip antenna has been fabricated to validate the simulated results, and its performance is tested experimentally. Additionally, the fidelity factor of face-to-face (FtF) and side-by-side (SbS) scenarios have been presented. The breast phantom models with a tumor and the antenna operating as a transceiver have been numerically simulated for the application of cancer tumor cell detection. The work will have a significant impact on the design of electromagnetic biosensors.

Vertical Graphene-Based Biosensor for Tumor Cell Dielectric Signature Evaluation.

The selective and rapid detection of tumor cells is of critical consequence for the theragnostic field of tumorigenesis; conventional methods, such as histopathological diagnostic methods, often require a long analysis time, excessive analytical costs, complex operations, qualified personnel and deliver many false-positive results. We are considering a new approach of an electrochemical biosensor based on graphene, which is evidenced to be a revolutionary nanomaterial enabling the specific and selective capture of tumor cells. In this paper, we report a biosensor fabricated by growing vertically aligned graphene nanosheets on the conductive surface of interdigitated electrodes which is functionalized with anti-EpCAM antibodies. The dielectric signature of the three types of tumor cells is determined by correlating the values from the Nyquist and Bode diagram: charge transfer resistance, electrical double layer capacity, Debye length, characteristic relaxation times of mobile charges, diffusion/adsorption coefficients, and variation in the electrical permittivity complex and of the phase shift with frequency. These characteristics are strongly dependent on the type of membrane molecules and the electromagnetic resonance frequency. We were able to use the fabricated sensor to differentiate between three types of tumor cell lines, HT-29, SW403 and MCF-7, by dielectric signature. The proposed evaluation method showed the permittivity at 1 MHz to be 3.63 nF for SW403 cells, 4.97 nF for HT 29 cells and 6.9 nF for MCF-7 cells.

Inflammatory pseudotumor-like follicular dendritic cell sarcoma with first clinical manifestation of thrombocytopenia: a case report

Inflammatory pseudotumor-like follicular dendritic cell sarcoma (IPT-like FDCS) is often associated with Epsrein-Barr (EB) virus infection. A male patient visited our hospital in September 2020 due to bleeding gums and epistaxis. The initial diagnosis was splenic lymphoma with marked thrombocytopenia. The patient was sent for pathological examination after splenectomy. Microscopic examination showed spindle-shaped or oval cells arranged in loose bundles, a large number of lymphocytes and plasma cells infiltrated in the interstitium, and fibrin-like changes in the blood vessel wall. Immunohistochemical detection of tumor cells was positive for CD21, CD35 and EBER, and we diagnosed IPT-like FDCS. The patient received platelet-raising therapy after operation. No tumor recurrence or metastasis was found during the 17-month follow-up, and the platelets returned to normal.IPT-like FDCS is an uncommon tumor, and the first presentation with marked thrombocytopenia is even rarer. The tumor was clinically and radiographically nonspecific. Its definitive diagnosis needs to rely on histopathological and immunohistochemical staining. IPT-like FDCS is biologically indolent and has a good prognosis.

427P Targeting adenosine (ADO) to improve the efficacy of combinations of cetuximab (CET) and immunotherapy in EGFRvIII MSS colorectal cancer (CRC)

The majority (∼85%) of CRCs that are microsatellite stable (MSS) are largely unresponsive to immunotherapy due to the lack of immune infiltration and low tumor mutation burden. Here, we demonstrated the value of EGFRvIII, a tumor-specific EGFR mutant, as the biomarker for patient stratification and explored therapeutic targets to convert cold tumors into responsive hot tumors. To identify EGFRvIII and assess immune infiltration, peripheral blood and tumor sections from newly diagnosed CRC patients and those receiving first-line CET-based chemotherapy were collected for circulating tumor cells (CTCs) detection and immunofluorescence (IF) staining. Sequencing data of CRC patients from TCGA was utilized for GSVA analysis to determine tumor immune microenvironment (TIME) phenotype. In total, EGFRvIII was identified in CTCs from 12% (14/115) of untreated CRC patients. Besides, 49 patients treated with CET were included in our analysis. The ORR of patients with baseline expression of EGFRvIII in CTCs (n=14) was decreased (14.3% vs 37.1%) accompanied with shorter TTP (median 5.5 vs 9.1 months, P=0.0086). CET resistance of MC38 EGFRvIII tumors diminished in immunodeficient nude mice, revealing the critical role of immune system in CET efficacy. GSVA analysis of TCGA MSS CRC cohort (n=192) showed that the TIME of EGFRvIII CRC was classified as immune-desert phenotype, characterized by the suppression of immunity. In CET treatment cohort, IF staining showed the significantly reduced infiltration of CD8+T cells and increased PD-L1 expression in EGFRvIII tumors compared with wild-type tumors. Subsequent experiments found that EGFRvIII CRC facilitated the conversion of pro-inflammatory extracellular ATP into immunosuppressive ADO, thus escaped from immunogenic cell death triggered by CET. Combined treatment of ADO pathway inhibitors restored the infiltration and anti-tumor activities of immune cells, indicating the potential application of second-line immune therapy in EGFRvIII MSS CRC. ADO-mediated suppressive TIME resulted in declined CET efficacy in EGFRvIII MSS CRC. Regulating ATP-to-ADO conversion remodeled TIME and may pave the way for immunotherapy.

1063P Comprehensive genomic profiling of leptomeningeal metastases on NSCLC patients through circulating tumor DNA in cerebrospinal fluid

Leptomeningeal metastases occurred in more than 3% of patients diagnosed with NSCLC, resulting in poor clinical outcomes. The cerebrospinal fluid (CSF) was a direct liquid biopsy for pathological diagnosis of leptomeningeal metastases. However, traditional clinical methods of detecting tumor cells in CSF showed limited sensitivity. Meanwhile, the unique genomic aberrations of leptomeningeal metastases remained unclear.

Novel Ultrasensitive Photoelectrochemical Cytosensor Based on Hollow CdIn2S4/In2S3 Heterostructured Microspheres for HepG2 Cells Detection and Inhibitor Screening.

Hepatocellular carcinoma is a life-threatening malignant tumor found around the world for its high morbidity and mortality. Therefore, it is of great importance for sensitive analysis of liver cancer cells (HepG2 cells) in clinical diagnosis and biomedical research. To fulfill this demand, hollow CdIn2S4/In2S3 heterostructured microspheres (termed CdIn2S4/In2S3 for clarity) were prepared by a two-step hydrothermal strategy and applied for building a novel photoelectrochemical (PEC) cytosensor for ultrasensitive and accurate detection of HepG2 cells through specific recognition of CD133 protein on the cell surface with the respective aptamer. The optical properties of CdIn2S4/In2S3 were investigated by UV-vis diffuse reflectance spectroscopy (DRS) and PEC technology. By virtue of their appealing PEC characteristics, the resultant PEC sensor exhibited a wider dynamic linear range from 1 × 102 to 2 × 105 cells mL-1 with a lower limit of detection (LOD, 23 cells mL-1), combined by evaluating the expression level of CD133 protein stimulated by metformin as a benchmarked inhibitor. This work opens a valuable and feasible avenue for sensitive detection of diverse tumor cells, holding great potential in early clinical diagnosis and treatment coupled by screening inhibitors.