Circulating tumor cells enrichment and single cell analysis in non-small cell lung cancer

Department of Biomedical EngineeringCompared to general-used tissue biopsy, less invasive ???liquid biopsy??? can provide insight which related to the real-time dynamics of cancer by more frequent analysis of circulating biomarkers including circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA). Furthermore, liquid biopsy is expected to offer a more comprehensive information of the disease, because circulating biomarkers may include cancer-associated materials from multiple disease sites in the body. However, the extreme rarity of cancer-associated circulating biomarkers in blood has been a great challenge to the achievement of the goals for liquid biopsy. CTC refers to tumor cell which disseminate from the primary tumor site to blood stream. Although CTC enumeration provides potential utility as a promising prognostic liquid biopsy marker, it has not been established yet due to its extreme rarity in comparison to other types of blood cells (1???10 CTCs/10^6 blood cells in 1 mL of blood). Representatively, CellSearch has been approved by the FDA about CTC enumeration system for breast, colorectal, and prostate cancer testing. And various microfluidic chips have been developed for CTC isolation based on physical and biochemical characteristics of CTCs. For example, some chips used epithelial cell adhesion molecule (EpCAM) antibody based capture and others used a size-selective manner. Many clinical papers were reported about the role of CTCs according to the remarkable improvement of the device. Notably, a high CTC count is associated with poor prognosis for cancer patients. Despite the clinical importance and progress of CTC-based cancer diagnostics, most of the current methods of enriching CTCs are difficult to implement in general hospital settings due to complex and time-consuming protocols. Among existing technologies, size-based isolation methods provide antibody-independent, relatively simple, and high throughput protocols. However, the clogging issues and lower than desired recovery rates and purity are the key challenges. In this thesis, fluid-assisted separation technology (FAST), which applied to centrifugal microfluidics with size-based membrane filtration, was suggested to solve clogging issues. FAST was inspired by antifouling membranes with liquid-filled pores in nature and it achieved high sensitivity (95.9 ?? 3.1% recovery rate) and high selectivity (>2.5 log depletion of white blood cells) without clogging. Moreover, rapid (>3 mL/min) isolation of viable CTCs from whole blood without prior sample treatment enabled simple and ultrafast process which could be applied in clinical field. Numerical simulation and experiments demonstrated that FAST disc achieved uniform, clog-free, ultrafast cell enrichment with pressure drops much less than in conventional size-based filtration, at 1 kPa. We validated the clinical utility of the point-of-care detection of CTCs with 142 cancer patients??? samples. For the depth study, we selected lung cancer to apply molecular analysis based on CTCs. In particularly, lung cancer is the most common cause of worldwide cancer-related mortality with 1.8 million new cases reported every year in both men and women. NSCLC accounts for 85% of all lung cancers and epidermal growth factor receptor (EGFR) mutations are the representative mutation in 10-30% of patients. According to the mutant type of NSCLC patients, targeted therapies including EGFR tyrosine kinase inhibitor (TKI) drugs can induce improved clinical outcomes. For obtaining molecular information about their tumors, detection of EGFR mutation from patient-driven CTCs should be validated through the confirmation of concordance with tissue biopsy results. Successive downstream analysis of single CTCs from each patient during treatment is needed for observing a change of characteristics related with therapeutic response. To make it possible, robust CTC isolation system with high efficiency is required. With the above needs in mind, we report the clinical validation and demonstration of a FAST disc capable of rapid CTC isolation and easy single cell picking for individual CTC characterization. We report here a proof-of-concept demonstration showing clinical meaning of CTCs included multigene expression as a monitoring marker in NSCLC.clos

Liquid biopsy in lung cancer: Clinical applications of circulating biomarkers (CTCs and ctDNA)

Lung cancer is by far the leading cause of cancer death worldwide, with non-small cell lung cancer (NSCLC) accounting for the majority of cases. Recent advances in the understanding of the biology of tumors and in highly sensitive detection technologies for molecular analysis offer targeted therapies, such as epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors. However, our understanding of an individual patient's lung cancer is often limited by tumor accessibility because of the high risk and invasive nature of current tissue biopsy procedures. Liquid biopsy, the analysis of circulating biomarkers from peripheral blood, such as circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA), offers a new source of cancer-derived materials that may reflect the status of the disease better and thereby contribute to more personalized treatment. In this review, we examined the clinical significance and uniqueness of CTCs and ctDNA from NSCLC patients, isolation and detection methods developed to analyze each type of circulating biomarker, and examples of clinical studies of potential applications for early diagnosis, prognosis, treatment monitoring, and prediction of resistance to therapy. We also discuss challenges that remain to be addressed before such tools are implemented for routine use in clinical settings

A lab-on-a-disc platform enables serial monitoring of individual CTCs associated with tumor progression during EGFR-targeted therapy for patients with NSCLC

Rationale: Unlike traditional biopsy, liquid biopsy, which is a largely non-invasive diagnostic and monitoring tool, can be performed more frequently to better track tumors and mutations over time and to validate the efficiency of a cancer treatment. Circulating tumor cells (CTCs) are considered promising liquid biopsy biomarkers; however, their use in clinical settings is limited by high costs and a low throughput of standard platforms for CTC enumeration and analysis. In this study, we used a label-free, high-throughput method for CTC isolation directly from whole blood of patients using a standalone, clinical setting-friendly platform. Methods: A CTC-based liquid biopsy approach was used to examine the efficacy of therapy and emergent drug resistance via longitudinal monitoring of CTC counts, DNA mutations, and single-cell-level gene expression in a prospective cohort of 40 patients with epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer. Results: The change ratio of the CTC counts was associated with tumor response, detected by CT scan, while the baseline CTC counts did not show association with progression-free survival or overall survival. We achieved a 100% concordance rate for the detection of EGFR mutation, including emergence of T790M, between tumor tissue and CTCs. More importantly, our data revealed the importance of the analysis of the epithelial/mesenchymal signature of individual pretreatment CTCs to predict drug responsiveness in patients. Conclusion: The fluid-assisted separation technology disc platform enables serial monitoring of CTC counts, DNA mutations, as well as unbiased molecular characterization of individual CTCs associated with tumor progression during targeted therapy

Characteristics predicting the efficacy of SGLT-2 inhibitors versus GLP-1 receptor agonists on major adverse cardiovascular events in type 2 diabetes mellitus: a meta-analysis study

Background Recent large clinical trials have demonstrated cardiovascular benefits of similar overall magnitude for sodium–glucose cotransporter-2 inhibitor (SGLT-2i) and glucagon-like peptide-1 receptor agonist (GLP-1RA) therapy in subjects with type 2 diabetes. We sought to identify subgroups based on baseline characteristics with a differential response to either SGLT-2i or GLP-1RA. Methods PubMed, Cochrane CENTRAL, and EMBASE were searched from 2008 to 2022 for SGLT-2i or GLP-1RA randomized trials that reported 3-point major adverse cardiovascular events (3P-MACE). Baseline clinical and biochemical characteristics included age, sex, body mass index (BMI), HbA1c, estimated glomerular filtration rate (eGFR), albuminuria, preexisting cardiovascular disease (CVD), and heart failure (HF). Absolute and relative risk reductions (ARR and RRR) regarding incidence rates for 3P-MACE with a 95% confidence interval were calculated. The association of average baseline characteristics in each study with the ARR and RRR for 3P-MACE was investigated by meta-regression analyses (random-effects model, assuming inter-study heterogeneity). Meta-analysis was also conducted to investigate whether the efficacy of SGLT-2i or GLP-1RA on 3P-MACE reduction could differ according to the patients characteristics (e.g., HbA1c above/below cutoff). Results After a critical assessment of 1,172 articles, 13 cardiovascular outcome trials with a total of 111,565 participants were selected. In meta-regression analysis, the more patients with reduced eGFR in the studies, the greater ARR by SGLT-2i or GLP-1RA therapy. Similarly, in the meta-analysis, SGLT-2i therapy tended to be more effective in reducing 3P-MACE in people with eGFR < 60 ml/min/1.73 m2 than in those with normal renal function (ARR − 0.90 [–1.44 to − 0.37] vs. − 0.17 [–0.34 to − 0.01] events/100 person-years). Furthermore, people with albuminuria tended to respond better to SGLT-2i therapy than those with normoalbuminuria. However, this was not the case for the GLP-1RA treatment. Other factors including age, sex, BMI, HbA1c, and preexisting CVD or HF did not affect the efficacy of either SGLT-2i or GLP-1RA treatment on the ARR or RRR of 3P-MACE. Conclusions Because decreased eGFR [significant] and albuminuria [trend] were found to predict a better efficacy for SGLT-2i in 3P-MACE reduction, this class of drug should be preferred in such patients. However, GLP-1RA may be considered for patients with normal eGFR because it showed better efficacy than SGLT-2i in this subgroup [trend]

Cause of microfibers found in the domestic washing process of clothing; focusing on the manufacturing, wearing, and washing processes

To prepare measures for washing synthetic fibers, which cause proliferation of microplastics in the marine ecosystem, a fundamental analysis is required. Therefore, this study established an efficient method for quantitatively analyzing microfibers using artificial neural networks, comparing the amounts of microfibers generated in the manufacturing, wearing, and washing processes of clothing. The proportion of microfiber emitted during the manufacturing process was the largest (49%), followed by that emitted during the washing (28%) and wearing (23%) processes. This suggests that minimizing the amount of microfiber emitted during the manufacturing process is key to solving microfiber issues in the fashion industry. Additionally, during the wearing process, the amount of waterborne microfiber detected in washing was slightly larger than the amount of airborne microfiber. In the washing process, the washing temperature did not significantly affect microfiber emissions. However, when reducing the amount of water used or increasing the number of washings, microfiber emissions increased noticeably due to the greater friction applied to clothes. A common result of all experiments was that the largest proportion of microfibers was released during the first five washing cycles. Therefore, before wearing new items, consumers can minimize microfiber release by pre-washing using a laundry bag that filters microfibers. Furthermore, the most effective way to minimize microfibers is to eliminate them from the manufacturing process before they are distributed to consumers.This study was conducted with the support of LG Electronics (No. 2-2020-1139-001-1)

Fifteen Years After the Gozan-Dong Glass Fiber Outbreak, Incheon in 1995

Objectives: In 1995, an outbreak survey in Gozan-dong concluded that an association between fiberglass exposure in drinking water and cancer outbreak cannot be established. This study follows the subjects from a study in 1995 using a data linkage method to examine whether an association existed. The authors will address the potential benefits and methodological issues following outbreak surveys using data linkage, particularly when informed consent is absent. Methods: This is a follow-up study of 697 (30 exposed) individuals out of the original 888 (31 exposed) participants (78.5%) from 1995 to 2007 assessing the cancer outcomes and deaths of these individuals. The National Cancer Registry (KNCR) and death certificate data were linked using the ID numbers of the participants. The standardized incidence ratio (SIR) and standardized mortality ratio (SMR) from cancers were calculated by the KNCR. Results: The SIR values for all cancer or gastrointestinal cancer (GI) occurrences were the lowest in the exposed group (SIR, 0.73; 95% CI, 0.10 to 5.21; 0.00 for GI), while the two control groups (control 1: external, control 2: internal) showed slight increases in their SIR values (SIR, 1.18 and 1.27 for all cancers; 1.62 and 1.46 for GI). All lacked statistical significance. All-cause mortality levels for the three groups showed the same pattern (SMR 0.37, 1.29, and 1.11). Conclusions: This study did not refute a finding of non-association with a 13-year follow-up. Considering that many outbreak surveys are associated with a small sample size and a cross-sectional design, follow-up studies that utilize data linkage should become standard procedure.OAIID:oai:osos.snu.ac.kr:snu2011-01/102/0000040632/15SEQ:15PERF_CD:SNU2011-01EVAL_ITEM_CD:102USER_ID:0000040632ADJUST_YN:YEMP_ID:A077602DEPT_CD:902CITE_RATE:0FILENAME:fifteen years after the gozan-dong glass fiber outbreak, incheon in 1995..pdfDEPT_NM:보건학과CONFIRM:

Circulating Tumor Cells Enumerated by a Centrifugal Microfluidic Device as a Predictive Marker for Monitoring Ovarian Cancer Treatment: A Pilot Study

We investigated the size-based isolation and enumeration of circulating tumor cells (CTCs) using a centrifugal microfluidic device equipped with a fluid-assisted separation technology (FAST) disc. We further assessed the correlations among CTCs, cancer antigen-125 (CA125) levels, and clinical course of the disease in a prospective analysis of 47 serial blood samples collected at multiple time-points from 13 ovarian cancer patients. CTCs were isolated from whole blood using the FAST disc and were classified as epithelial cell adhesion molecule (EpCAM)/cytokeratin+, CD45-, and 4 &apos;,6-diamidino-2-phenylindole (DAPI)+. Mean CTC count at baseline was 20.2; 84.62% of patients had more than one CTC at baseline and had decreased CTCs counts after surgery and chemotherapy. The CTC counts in eight patients with complete responses were &lt;3. CTC counts were correlated with CA125 levels in three patients without recurrence; they were elevated in three patients with recurrence and normal CA125 concentrations. CTC counts and CA125 levels showed high concordance with directional changes (increasing 71.4%; non-increasing 75.0%). CTC counts showed higher associations with clinical status, sensitivity (100.0% vs. 60.0%), positive predictive value (55.6% vs. 42.9%), and negative predictive value (100.0% vs. 87.5%) than CA125 levels. CTC counts were better associated with treatment response and recurrence than CA125 levels

Breath analyzer for personalized monitoring of exercise-induced metabolic fat burning

Dionisio V. Del Orbe recibió su Licenciatura en Ingeniería Aeronáutica de la Universidad de Western Michigan (2012), EE. UU., y una Maestría en Ingeniería de Manufactura Microelectrónica del Instituto de Tecnología de Rochester (2015), EE. UU. Recibió su doctorado en Ingeniería Mecánica KAIST (2022), Corea del Sur, y trabajó como investigador de posgrado en el Departamento de Investigación de TIC Médicas y de Bienestar en ETRI, Corea del Sur. Su investigación se centra en sensores de gases químicos para diversas aplicaciones, especialmente, análisis de aliento y detección de gases tóxicos/inflamables; también tiene intereses en dispositivos portátiles y flexibles. Actualmente, es docente e investigador en UNAPEC, República Dominicana.Obesity increases the risk of chronic diseases, such as type 2 diabetes mellitus, dyslipidemia, and cardiovascular diseases. Simple anthropometric measurements have time limitations in reflecting short-term weight and body fat changes. Thus, for detecting, losing or maintaining weight in short term, it is desirable to develop portable/ compact devices to monitor exercise-induced fat burn in real time. Exhaled breath acetone and blood-borne β-hydroxybutyric acid (BOHB) are both correlated biomarkers of the metabolic fat burning process that takes place in the liver, predominantly post-exercise. Here, we have fabricated a compact breath analyzer for convenient, noninvasive and personalized estimation of fat burning in real time in a highly automated manner. The analyzer collects end-tidal breath in a standardized, user-friendly manner and it is equipped with an array of four low-power MEMS sensors for enhanced accuracy; this device presents a combination of required and desirable design features in modern portable/compact breath analyzers. We analyzed the exhaled breath (with our analyzer) and the blood samples (for BOHB) in 20 participants after exercise; we estimated the values of BOHB, as indication of the fat burn, resulting in Pearson coefficient r between the actual and predicted BOHB of 0.8. The estimation uses the responses from the sensor array in our analyzer and demographic and anthropo- metric information from the participants as inputs to a machine learning algorithm. The system and approach herein may help guide regular exercise for weight loss and its maintenance based on individuals’ own metabolic changes

Circulating Tumor Cell Clusters Are Cloaked with Platelets and Correlate with Poor Prognosis in Unresectable Pancreatic Cancer

Simple Summary: Despite recent advances, some patients with pancreatic cancer are refractory to treatment and the disease rapidly progresses, resulting in early death. The potential prognostic value of circulating tumor cells (CTCs) has been demonstrated in other cancer types, but the clinical validity in pancreatic cancer remains elusive. Here, we show that CTC clusters, which show mesenchymal characteristics and platelet marker expression, are highly correlated with poor prognosis in patients with unresectable pancreatic cancer.Circulating tumor cells (CTCs) are known to be heterogeneous and clustered with tumor-associated cells, such as macrophages, neutrophils, fibroblasts, and platelets. However, their molecular profile and clinical significance remain largely unknown. Thus, we aimed to perform a comprehensive gene expression analysis of single CTCs and CTC clusters in patients with pancreatic cancer and to identify their potential clinical relevance to provide personalized medicine. Epitope-independent, rapid (&gt; 3 mL of whole blood/min) isolation of single CTCs and CTC clusters was achieved from a prospective cohort of 16 patients with unresectable pancreatic cancer using a centrifugal microfluidic device. Forty-eight mRNA expressions of individual CTCs and CTC clusters were analyzed to identify pancreatic CTC phenotype. CTC clusters had a larger proportion of mesenchymal expression than single CTCs (p = 0.0004). The presence of CTC clusters positively correlated with poor prognosis (progression-free survival, p = 0.0159; overall survival, p = 0.0186). Furthermore, we found that most CTCs in these patients (90.7%) were cloaked with platelets and found the presence of a positive correlation between the increase in CTC clusters and rapid disease progression during follow-ups. Efficient CTC cluster isolation and analysis techniques will enhance the understanding of complex tumor metastasis processes and can facilitate personalized disease management