Microfluidic Assaying of Circulating Tumor Cells and Its Application in Risk Stratification of Urothelial Bladder Cancer

Bladder cancer is characterized by its frequent recurrence and progression. Effective treatment strategies need to be based on an accurate risk stratification, in which muscle invasiveness and tumor grade represent the two most important factors. Traditional imaging techniques provide preliminary information about muscle invasiveness but are lacking in terms of accuracy. Although as the gold standard, pathological biopsy is only available after the surgery and cannot be performed longitudinally for long-term surveillance. In this work, we developed a microfluidic approach that interrogates circulating tumor cells (CTCs) in the peripheral blood of bladder cancer patients to reflect the risk stratification of the disease. In a cohort of 48 bladder cancer patients comprising 33 non-muscle invasive bladder cancer (NMIBC) cases and 15 muscle invasive bladder cancer (MIBC) cases, the CTC count was found to be considerably higher in the MIBC group compared with the NMIBC group (4.67 vs. 1.88 CTCs/3 mL, P=0.019), and was significantly higher in high-grade bladder cancer patients verses low-grade bladder cancer patients (3.69 vs. 1.18 CTCs/3mL, P=0.024). This microfluidic assay of CTCs is believed to be a promising complementary tool for the risk stratification of bladder cancer

Evaluation of Circulating Endometrial Cells as a Biomarker for Endometriosis

Background: Circulating endometrial cells (CECs) have been reported to be present in the peripheral blood of women with endometriosis (EM), providing clear and specific evidence of the presence of ectopic lesions. In this study, we established a method with a high detection rate of CECs, assessed the diagnostic value of CECs for EM and compared with serum CA125, and proposed a hypothesis for the pathogenesis of EM from the new perspective of CECs. Methods: The participants were enrolled prospectively from October 2015 to July 2016. The peripheral blood samples were collected from 59 participants, and the blood cells were isolated for immunofluorescence staining via microfluidic chips. The cells that were positive for vimentin/cytokeratin and estrogen/progesterone receptor and negative for CD45 were identified as CECs. The serum CA125 level was tested with electrochemiluminescence immunoassay. Results: The detection rate of CECs reached 89.5% (17/19) in the EM group, which was significantly higher than that of the control group (15.0% [6/40], P < 0.001) and was independent of menstrual cycle phases. Furthermore, a positive CEC assay detected 4/5 cases of Stage I–II EM. In contrast, a positive CA125 test had limited value in detecting EM (13/19, 68.4%) and detected only one case of Stage I–II EM. Conclusion: CECs are promising biomarkers for EM with great potential for a noninvasive diagnostic assay

Microfluidic assay of circulating endothelial cells in coronary artery disease patients with angina pectoris - Fig 4

<p>(<b>A</b>) Immunofluorescence staining of isolated CECs. Captured CECs (DAPI⁺/CD146⁺/VEGFR1⁺/CD45^-) can be clearly observed at the single cell level, together with residuary leukocytes (DAPI⁺/CD45⁺). (<b>B</b>) Comparative CECs counts among HC group, CSA group and UA group. Data are expressed as the median with IQRs. (<b>C</b>) Comparative CEC counts with increasing TIMI UA/NSTEMI risk score. Data are expressed as the median with IQRs. P-value referred to the results of Mann-Whitney test between groups. Scale bar, 20μm.</p

Calculation of risk score using the TIMI NSTEMI/UA system.

<p>Calculation of risk score using the TIMI NSTEMI/UA system.</p

Correlations between CEC count and typical cardiac biomarkers.

<p>Spearman rank correlation coefficient (<i>ρ</i>) was used to assessed the correlation between CEC counts in CAD patients with angina pectoris and each initial presenting serum cardiac biomarkers. (<b>A</b>) No correlation was observed between CEC count and cTnI value (<i>ρ</i> = 0.01, <i>p</i> = 0.987). (<b>B</b>) No correlation was observed between CEC count and AST value (<i>ρ</i> = 0.02, <i>p</i> = 0.910). (<b>C</b>) No correlation was observed between CEC count and LDH value (<i>ρ</i> = 0.15, <i>p</i> = 0.271). (<b>D</b>) No correlation was observed between CEC count and CK value (<i>ρ</i> = 0.18, <i>p</i> = 0.181). (<b>E</b>) No correlation was observed between CEC count and CK-MB value (<i>ρ</i> = -0.14, <i>p</i> = 0.326). (<b>F</b>) No correlation was observed between CEC count and α-HBDH value (<i>ρ</i> = 0.17, <i>p</i> = 0.212).</p

Baseline characteristics and CEC count for the three comparison groups.

<p>Baseline characteristics and CEC count for the three comparison groups.</p

Receiver-operator characteristic curves illustrating the predictive accuracy of CEC count for the diagnosis of CAD with angina pectoris.

<p>(<b>A</b>) AUC was equal to 0.867 (<i>p</i> < 0.01). The red point represented a classification cutoff value of 2.7 CEC/ml, which is associated with a sensitivity of 87.5% and specificity of 66.7% to accurately differentiate a CSA case from a healthy control. (<b>B</b>) AUC was equal to 0.938 (<i>p</i> < 0.001). The red point represented a classification cutoff value of 4.2 CEC/ml, which is associated with a sensitivity of 84.6% and specificity of 86.7% to accurately differentiate a UA case from a healthy control.</p

Comparing CEC count with an increasing TIMI UA/NSTEMI risk score for the 3 categories.

<p>Comparing CEC count with an increasing TIMI UA/NSTEMI risk score for the 3 categories.</p

Size distribution of HUVECs.

<p>Min: 16.8μm, max: 35.3μm, mean: 24.0μm, SD: 3.2μm, skewness: -0.072 and kurtosis: 0.882.</p