Benign Lung Disease Research Articles

A novel CDA liquid biopsy technology for effective non-small cell lung cancer diagnosis.

e23131 Background: Cancer Differentiation Analysis liquid biopsy (CDA) has been investigated as a viable clinical utility in NSCLC diagnosis, as well as a new candidate for part of an overall post-CT scan clinical diagnostic and treatment decision tool kit. It has been also evaluated in NSCLC diagnosis, combined with imaging technologies. Methods: This was a retrospective investigation in which 160 individuals were recruited at Changhai Hospital of Shanghai from July to Dec. in 2014. CDA and CT scan tests were performed on all samples before final confirmation by biopsy. For CDA tests, peripheral blood was drawn in EDTA tubes before operation. A performance predication model of CDA and CT scan test results was built using pROC package in R Language for the data of Area Under the Curve (AUC) and CDA threshold values. Further analysis was carried out based on the CDA threshold values. Results: Out of the 160 individuals, 40 were diagnosed as benign lung diseases selected as control group, 120 were confirmed as NSCLC by pathology. CDA and CT test data along with epidemiological information were collected and complied. Details of other statistical results were given in the Table below. Conclusions: CDA liquid biopsy is a novel, multi-level, multi-parameter based disease diagnosis technology with improvements in multiple areas including but not limited to higher sensitivity and specificity, ability to diagnose cancer early, no side effects, and covering a wide range of cancer sites. Results showed that AUC values of CDA were higher than those of CT scan for all NSCLC stage groups. CDA plus CT scan combination has the highest average AUC value for all stages. CDA test can provide more accurate and reliable diagnostic information and data for oncologists in making crucial clinical decisions. It is expected that CDA liquid biopsy technology will play an important and critical role in NSCLC diagnosis. [Table: see text]

Systematic review and meta-analysis of the efficacy of serum neuron-specific enolase for early small cell lung cancer screening.

We performed a pooled analysis of the efficacy of serum neuron-specific enolase (NSE) levels for early detection of small cell lung cancer (SCLC) in patients with benign lung diseases and healthy individuals. Comprehensive searches of several databases through September 2016 were conducted. The quality of the included studies was assessed using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) tool. Ultimately, 33 studies containing 9546 samples were included in the review. Pooled sensitivity of NSE for detecting SCLC was 0.688 (95%CI: 0.627-0.743), specificity was 0.921 (95%CI: 0.890-0.944), positive likelihood ratio was 8.744 (95%CI: 6.308-12.121), negative likelihood ratio was 0.339 (95%CI: 0.283- 0.405), diagnostic odds ratio was 25.827 (95%CI: 17.490- 38.136) and area under the curve was 0.88 (95%CI: 0.85- 0.91). Meta-regression indicated that study region was a source of heterogeneity in the sensitivity and joint models, while cut-off level was a source in the joint model. Subgroup analysis showed that enzyme linked immunosorbent assays had the highest sensitivity and radioimmunoassay assays had the highest specificity. The diagnostic performance was better in Europe [sensitivity: 0.740 (95%CI: 0.676-0.795), specificity: 0.932 (95%CI: 0.904-0.953)] than in Asia [sensitivity: 0.590 (95%CI: 0.496- 0.678), specificity: 0.901 (95%CI: 0.819-0.948)]. In Europe, 25 ng/ml is likely the most suitable NSE cut-off level. NSE thus has high diagnostic efficacy when screening for SCLC, though the efficacy differs depending on study region, assay method and cut-off level. In the clinic, NSE measurements should be considered along with clinical symptoms, image results and histopathology.

E2F3 transcription factor: A promising biomarker in lung cancer.

Many researches aiming to explore the pathogenesis of lung cancer have extensively studied the molecular alteration in such disease. In the present study we measured the blood E2F3 mRNA using real-time RT-PCR technique in order to evaluate its clinical significance in early diagnosis and monitoring of lung cancer. This case-control study included 50 lung cancer patients, 20 patients with benign lung diseases and 20 healthy controls. Relative quantification of blood E2F3 mRNA was done by real-time RT-PCR. Blood E2F3 mRNA levels were significantly higher in lung cancer patients when compared to either patients with benign lung diseases or healthy subjects. This elevation was significant in those with metastatic lung cancer as compared to those with localized lung cancer. At a cutoff^{(2-Δ Δ CT)} 1.5, blood E2F3 mRNA was able to distinguish malignant from benign lung conditions with a diagnostic sensitivity of 100%; while at a cutoff^{(2-Δ Δ CT)} 5.3, blood E2F3 mRNA discriminated localized from metastatic lung cancer with a sensitivity of 93.6%. Blood E2F3 mRNA is a sensitive diagnostic marker in lung cancer; moreover, it is a promising prognostic marker capable of efficiently discriminating early from late stages of the disease.

Predictive and prognostic value of serum periostin in advanced non-small cell lung cancer patients receiving chemotherapy.

Periostin is an extracellular matrix protein involved in tumorigenesis and metastasis. However, the role of serum periostin as a surrogate marker for treatment efficacy is still unknown. In 122 advanced non-small cell lung cancer cases, 37 patients with benign lung disease and 40 healthy controls, serum periostin was measured by enzyme-linked immunosorbent assays. The associations of serum periostin levels with the clinic-pathological parameters, chemotherapy response, and clinical outcomes of non-small cell lung cancer patients were analyzed. Serum periostin levels were significantly higher in non-small cell lung cancer patients, and it was related significantly to bone metastasis ( p = 0.021). Serum periostin of 65 non-small cell lung cancer patients were detected before and after two cycles of chemotherapy. The patients with and without periostin response had significant difference in objective response to chemotherapy ( p = 0.001). For the 122 non-small cell lung cancer patients, the median progression-free survival was 5 months. In a multivariate analysis, performance status (hazard ratio, 1.71; 95% confidence interval, 1.10-2.67), baseline periostin (hazard ratio, 1.01; 95% confidence interval, 1.00-1.01), and periostin response (hazard ratio, 0.50; 95% confidence interval, 0.29-0.86) were significantly correlated with prognosis. In conclusion, serum periostin was elevated in advanced non-small cell lung cancer patients. Baseline periostin and periostin responses appeared to be reliable surrogate markers to predict chemotherapy response and survival in patients with advanced non-small cell lung cancer.

Clinical Significance of Folate Receptor-positive Circulating Tumor Cells Detected by Ligand-targeted Polymerase Chain Reaction in Lung Cancer.

Background: As the heterogeneity of CTCs is becoming increasingly better understood, it is clear that identifying particular subtypes of CTCs would be more relevant.Methods: We detected folate receptor (FR)-positive circulating tumor cells (FR+-CTCs) by a novel ligand-targeted polymerase chain reaction (LT-PCR) detection technique.Results: In the none-dynamic study, FR+-CTC levels of patients with lung cancer were significantly higher than controls (patients with benign lung diseases and healthy controls). With a threshold of 8.7 CTC units, FR+-CTC showed a sensitivity of 77.7% and specificity of 89.5% in the diagnosis of lung cancer. When compared with established clinical biomarkers including carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA21-1), and neuron-specific enolase (NSE), FR+-CTC showed the highest diagnostic efficiency. Notably, the combination of FR+-CTC, CEA, NSE, and CYFRA21-1 could significantly improve the diagnostic efficacy in differentiating patients with lung cancer from benign lung disease. In our dynamic surveillance study, the CTC levels of 62 non-small cell lung cancer (NSCLC) patients decreased significantly after tumor resection.Conclusion: We established a LT-PCR-based FR+-CTC detection platform for patients with lung cancer that exhibits high sensitivity and specificity. This platform would be clinical useful in lung cancer diagnosis and treatment response assessment.

Pulmonary actinomycosis mimicking lung cancer on positron emission tomography.

Pulmonary actinomycosis frequently mimics lung malignancy on radiologic imaging studies. Positron emission tomography-computed tomography (PET-CT) is a useful diagnostic modality for differentiating lung malignancy from benign diseases. However, few studies evaluated PET-CT findings of pulmonary actinomycosis. Therefore, it is unclear whether PET-CT is helpful to distinguish lung malignancy from benign lung disease when pulmonary actinomycosis is clinically suspected. We investigated PET-CT findings in 11 patients with pathologically confirmed pulmonary actinomycosis. The median maximal standardized uptake value (SUV) on PET-CT of pulmonary actinomycosis was increased to 5.5 (interquartile range, 4.2–8.8), which was higher than the threshold value of 2.5 indicating malignancy. Pulmonary actinomycosis without central necrosis demonstrated higher maximal SUV of 7.5 (4.9–12.2) compared to 4.8 (3.2–5.6) of ones with central necrosis. PET-CT might be not helpful in differentiating lung malignancy from benign lesions when pulmonary actinomycosis is clinically suspected.

DNA Methylation Analysis of the SHOX2 and RASSF1A Panel in Bronchoalveolar Lavage Fluid for Lung Cancer Diagnosis.

Introduction: Currently the majority of lung cancer patients are diagnosed as advanced diseases for no sensitive and specific biomarkers exist, noninvasive biomarkers with high sensitivity and specificity are urgently needed in lung cancer diagnosis. Bronchoscopy is a standard procedure of the diagnostic work-up of patients with suspected lung cancer despite of the limited diagnostic accuracy. Besides, epigenetic changes through DNA methylation play an important role in tumorigenesis. Thus, we examined the aberrant methylation of the SHOX2 and RASSF1A in bronchoalveolar lavage fluid (BALF) in comparing with conventional cytology examination and serum CEA in order to evaluate the new diagnostic method.Patients and Methods: BALF and serum samples were collected from 322 patients at the time of diagnosis, 284 of them were pathologically confirmed lung cancer, 35 were benign lung diseases and 3 were malignancies in other systems. For all of the 322 patients, the methylation status of the SHOX2 and RASSF1A gene were detected by a new RT-PCR platform and then confirmed by sanger sequencing. Serum CEA were detected using electrochemiluminescence immunoassay.Results: Profiling data showed the consistency of RT-PCR and sanger sequencing in detecting the methylation of the SHOX2 and RASSF1A. Besides, the combination of SHOX2 and RASSF1A methylation in BALF yielded a diagnostic sensitivity of 81.0% and specificity of 97.4%. When compared with established cytology examination (sensitivity: 68.3%, specificity: 97.4%) and serum biomarker carcinoembryonic antigen (CEA) (sensitivity: 30.6%, specificity: 100.0%), the SHOX2 and RASSF1A methylation panel showed the highest diagnostic efficiency. Notably, the combination of cytology and the SHOX2 and RASSF1A methylation panel could significantly improve the diagnostic efficacy.Conclusion: The methylation analysis of the SHOX2 and RASSF1A panel in BALF with RT-PCR achieved a satisfactory sensitivity and specificity in lung cancer diagnosis, especially in an early stage. It could be used as a promising noninvasive biomarker for auxiliary diagnosis of lung cancer.

Significance of Serum MicroRNAs in the Auxiliary Diagnosis of Non-Small Cell Lung Cancer.

The lack of effective means for the early diagnosis of non-small cell lung cancer (NSCLC) is the leading cause of the high mortality associated with this form of lung cancer. This study aims to explore the potential significance of serum miRNA in the auxiliary diagnosis of NSCLC. The relative serum levels of 10 miRNAs in 120 patients with NSCLC, 45 patients with benign lung diseases, and 45 healthy controls were detected using real-time quantitative polymerase chain reaction (PCR). The receiver operating characteristic (ROC) curves were then used to analyze the significance of the expression of these 10 miRNAs in the diagnosis of NSCLC, as well as to compare them with the current commonly used tumor marker carcinoembryonic antigen (CEA). The serum levels of miR-125b and miR-22 in the NSCLC patients were significantly higher than those in the other two groups (p < 0.05), but the serum expression of miR-15b in the NSCLC patients was significantly lower than that in the other two groups (p < 0.01). The sensitivities of serum miR-22 and miR-15b in detecting early NSCLC (stage I + II) were significantly higher than that of CEA (p < 0.05). Area under the curves (AUCs) of serum miR-22, miR-125b, and miR-15b in the diagnosis of NSCLC were 0.725, 0.704, and 0.619, respectively, and the diagnostic significance of these three serum miRNAs for NSCLC was higher than that of serum CEA (AUC = 0.594). Serum miRNAs have potential as NSCLC-screening tumor markers, and serum miR-22 and miR15b might be used as reference indexes for the early diagnosis of NSCLC in the future.

Simultaneous Quantification of Serum Multi-Phospholipids as Potential Biomarkers for Differentiating Different Pathophysiological states of lung, stomach, intestine, and pancreas.

Background: Aberrant lipid metabolism is closely associated with cancer.Materials & Methods: Serum levels of sphingomyelins (SM) (34:1), phosphatidylcholine (PC) (34:2), PC(34:1), PC(36:4), PC(36:3), and PC(36:2) in 1449 serum samples (including 599 normal controls, 69 patients with benign lung diseases (BLDs), 61 with benign colorectal diseases, 54 with benign gastric diseases, 67 with benign pancreatic diseases, and 246 with lung cancer (LC), 144 with colorectal cancer, 94 with gastric cancer, 115 with pancreatic cancer) were quantified simultaneously based on their respective calibration equations with correlation coefficient of >0.98.Results: Receiver operating characteristic (ROC) analysis indicated that 18 panels obtained from these six phospholipids have high diagnostic ability to differentiate between different pathophysiological states. For example, a combination of SM(34:1), PC(34:2), PC(34:1), PC(36:3), and PC(36:2) to differentiating male patients with early stage LC from male normal controls plus male BLDs with a value under ROC curve (AUC) of 0.957, a sensitivity of 88.9%, and a specificity of 90.8%. SM(34:1) and PC(34:1) to differentiating female patients with early stage LC from female normal controls plus female BLDs with an AUC of 0.903, a sensitivity of 90.0%, and a specificity of 77.5%.Conclusion: Change trends of these six phospholipids were significantly correlated with gender, physiological states, and cancer stages.

P1.05-022 Circulating Tumor Cell Isolation to Monitor NSCLC Patients over the Course of Treatment

Compared to the investigation of the primary tumor or a biopsy taken from a distant metastasis, the investigation of a patient’s blood is relatively simple, less invasive and can be performed repeatedly. Thus, CTC (circulating tumor cells) investigation can be used as a real-time marker for staging, disease progression and therapy responsiveness. Cancer mortality might be reduced dramatically when the disease and its metastatic spread are detected and characterized early and can therefore be treated in an optimal fashion. The GILUPI CellCollector® offers medical personnel at any point-of-care with the unique opportunity to enrich these CTCs in vivo. Here, we conducted a study using this effective device, to monitor CTC counts before as well as on different time points after surgery in non-small cell lung cancer (NSCLC) patients and further to characterize the CTCs on a molecular level. In total, 20 NSCLC patients (different stages) were screened for CTCs at different time points: preoperative, 30 minutes after tumor resection, 1 week postoperative as well as in 3-monthly intervals up to 2 years. In addition, 1 patient with a benign lung disease were included in this study. CTCs were isolated independent from tumor stages and even in quite early cases CTCs could be detected. Moreover, a difference between CTC occurrence before and after surgery was seen and a correlation between CTC enumeration and clinical lack of recurrence could be detected. The GILUPI CellCollector® overcomes blood volume limitations of other diagnostic approaches and thereby increases the diagnostic sensitivity of CTC analysis. Future implementation into clinical practice may improve early detection, prognosis and therapy monitoring of cancer patients. Besides enumeration, captured CTCs are ready for molecular characterization and will help to establish more personalized treatment regiments since knowledge of the molecular make-up of the cancer cells to be defeated is an indispensable prerequisite to use targeted therapies efficiently.

P2.01-073 The Diagnostic Value of Carcinoembryonic Antigen and Squamous Cell Carcinoma Antigen in Lung Adenosquamous Carcinoma

Lung adenosquamous carcinoma (ASC) is a rare malignant tumor with an adenocarcinoma and a squamous cell carcinoma component, and associated with a lower 5-year survival rate than lung squamous cell carcinoma and lung adenocarcinoma. Surgical specimen histology revealed inadequacy of conventional transbronchial needle aspiration sample in the diagnosis of lung ASC. Most of lung ASC patients are not suitable to receive surgery, and it is difficult to diagnosis ASC. This study is to explore the possibility of using serum carcinoembryonic antigen (CEA) and squamous cell carcinoma antigen (SCC) as a supplementary diagnostic test for ASC. We retrospectively analyzed the preoperative serum CEA and SCC levels in 34 patients with lung ASC, 35 cases of lung adenocarcinoma patients, 35 cases of lung squamous cell carcinoma patients. 36 cases of lung benign disease patients and 35 cases of healthy people as control group were also retrospectively collected and analyzed from January 2012 to December 2014 at the Zhejiang Cancer Hospital, China. The differences of CEA and SCC among the groups were evaluated, and the area under the curve (AUC), sensitivity and specificity were calculated. The levels of SCC and CEA in lung ASC group were significantly higher than those in healthy control group and benign disease group (P<0.05), and SCC level in lung ASC group was significantly higher than that in lung adenocarcinoma group (P<0.05). CEA and SCC had good diagnostic sensitivity and specificity compared with the healthy control group, and the difference was statistically significant (P<0.05). Our retrospective study suggested a role for serum CEA and SCC levels as reference markers in the diagnosis of lung ASC. If the patients which CEA and SCC levels were elevated and diagnosed as lung adenocarcinoma by limited biopsy materials should be offered further work-up to reach an accurate diagnosis and treatment.

The Diagnostic Value of Carcinoembryonic Antigen and Squamous Cell Carcinoma Antigen in Lung Adenosquamous Carcinoma.

Lung adenosquamous carcinoma (ASC) is a rare malignant tumor with an adenocarcinoma and a squamous cell carcinoma component and associated with a lower 5-year survival rate than lung squamous cell carcinoma and lung adenocarcinoma. Surgical specimen histology revealed the inadequacy of conventional transbronchial needle aspiration samples in the diagnosis of lung ASC. Most lung ASC patients are not suitable to receive surgery, and it is difficult to diagnose ASC. This study is to explore the possibility of using serum carcinoembryonic antigen (CEA) and squamous cell carcinoma antigen (SCC) as a supplementary diagnostic test for ASC. We retrospectively analyzed the preoperative serum CEA and SCC levels in 34 patients with lung ASC, 35 cases of lung adenocarcinoma patients, 35 cases of lung squamous cell carcinoma patients. 36 cases of lung benign disease patients and 35 cases of healthy people as a control group were also retrospectively collected and analyzed from January 2012 to December 2014 at the Zhejiang Cancer Hospital, China. The differences of CEA and SCC among the groups were evaluated, and the area under the curve (AUC), sensitivity, and specificity were calculated. The levels of SCC and CEA in the lung ASC group were significantly higher than those in the healthy control group and benign disease group (p < 0.05). The SCC level in lung ASC group was significantly higher than that in lung adenocarcinoma group (p < 0.05). CEA and SCC had good diagnostic sensitivity and specificity compared with the healthy control group, and the difference was statistically significant (p < 0.05). Our retrospective study suggested a role for serum CEA and SCC levels as reference markers in the diagnosis of lung ASC. Patients with elevated CEA and SCC levels and diagnosed as lung adenocarcinoma by limited biopsy materials should be offered further work-up to reach an accurate diagnosis and treatment.

Diagnostic Value of Folate Receptor-positive Circulating Tumor Cell in Lung Cancer: A Pilot Study

背景与目的评价一种通过叶酸受体(folate receptor, FR)检测循环肿瘤细胞(circulating tumor cell, CTC)的方法用于肺癌临床诊断的实用性和可行性及进一步探究CTC在肺癌术后复发的预测价值。方法通过免疫磁珠负向富集方法从3 mL外周血中捕获循环肿瘤细胞，再用肿瘤特异性叶酸配体-寡核苷酸偶和物标记捕获的循环肿瘤细胞，洗去没有结合的偶和物后，洗脱下特异性结合的偶合物的寡核苷酸用于定量PCR扩增分析。结果97例肺癌患者的CTC水平高于肺部良性疾病患者(P < 0.001)。本检测方法以8.7 Folate Units/3 mL为cutoff值，结果显示靶向PCR法对肺癌的检测灵敏度为82.5%，特异性为72.2%，特别是在Ⅰ期肺癌灵敏度达到86.8%。与其他肿瘤标志物(NSE、CEA、CYFRA21-1)比较，CTC对肺癌及Ⅰ期肺癌具有较高的诊断准确性(0.859; 95%CI: 0.779-0.939)和(0.912; 95%CI: 0.829-0.994)。5例肺癌患者术后2周内CTC水平高于cutoff值。结论叶酸受体阳性循环肿瘤细胞可以应用于肺癌的临床诊断，即使是对早期非小细胞肺癌(non-small cell lung cancer, NSCLC)的诊断。

Diagnostic and prognostic value of serum periostin in patients with non-small cell lung cancer.

The periostin protein is expressed in a variety of human malignancies. The aim of this study was to explore the diagnostic and prognostic value of serum periostin levels in patients with non-small cell lung cancer (NSCLC). We measured serum periostin levels by ELISA in 296 NSCLC patients, 120 benign lung diseases (BLD) patients and 160 healthy controls. The levels of serum periostin in NSCLC patients were significantly elevated compared with those in healthy controls (P < 0.001) and BLD patients (P < 0.001). Using a cutoff value of 30.87 ng/ml, the sensitivity and specificity of periostin in differentiating between NSCLC patients and BLD patients, and between NSCLC patients and healthy controls was, 48.6 and 91.7%, and 51.4 and 97.5%, respectively. Kaplan-Meier log rank analysis revealed that the higher serum periostin levels group had a poorer progression-free survival (PFS) and overall survival (OS) compared with lower periostin group (P = 0.024, P = 0.015, respectively). Further univariate and multivariate Cox regression analysis showed that serum periostin was an independent risk factor of prognosis of NSCLC patients. In conclusion, our study suggests that serum periostin could be considered as a diagnostic and prognostic marker for NSCLC patients.

Glycodelin is a potential novel follow-up biomarker for malignant pleural mesothelioma.

Malignant pleural mesothelioma (MPM) is a rare and aggressive tumor with a short survival time arising from the mesothelial cells of the pleura. Soluble mesothelin-related peptide (SMRP), osteopontin or EFEMP1 (Fibulin-3) are well described biomarkers for malignant mesothelioma with moderate sensitivity and specificity. In this study, we characterized the expression of glycodelin, a marker for risk pregnancy, in MPM by RNA and protein analyses and investigated its potential as a MPM biomarker. We were able to detect glycodelin in the serum of MPM patients. Compared to benign lung diseases, the serum levels were significant increased. Patients with high glycodelin serum levels revealed a worse overall survival. The glycodelin serum levels correlated with the tumor response to treatment. A comparison of SMRP and glycodelin serum measurement in a large patient cohort demonstrated that the detection of both soluble factors can increase the reliable diagnostic of MPM. Glycodelin was highly expressed in MPM tumors. Analyses of a tissue micro array indicated that the immunomodulatory form glycodelin A was expressed in MPM and correlated with the survival of the patients. Altogether, glycodelin seems to be a new potential biomarker for the aggressive malignant pleural mesothelioma.

Simultaneous Quantification of Serum Nonesterified and Esterified Fatty Acids as Potential Biomarkers to Differentiate Benign Lung Diseases from Lung Cancer.

In this study, we have employed graphene oxide as a matrix to simultaneously and directly quantify serum nonesterified and esterified fatty acids (FAs) using matrix-assisted laser/desorption ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS). Twelve serum nonesterified FAs combined with their individual esterified FAs (i.e., C16:0, C16:1, C18:0, C18:1, C18:2, C18:3, C20:2, C20:3, C20:4, C20:5, C22:5, and C22:6) were quantified based on their calibration curves with the correlation coefficients of >0.99, along with the analytical time of <1 min each sample. As a result, serum levels of twelve total FAs (TFAs) in 1440 serum samples from 487 healthy controls (HCs), 479 patients with benign lung diseases (BLDs) and 474 patients with lung cancer (LC) were determined. Statistical analysis indicated that significantly increased levels of C16:0, C16:1, C18:0, C18:1, C18:3, C20:3, and C22:6 and decreased levels of C20:5 were observed in LC patients compared with BLDs. Receiver operating characteristic (ROC) analysis revealed that panel a (C18:2, C20:3, C20:4, C20:5, C22:5, and C22:6), panel b (C18:0, C20:4, C20:5, and C22:6), and panel c (C16:1, C18:0, C18:1, C20:3, and C22:6) have exhibited good diagnostic ability to differentiate BLDs from LC relative to clinical uses of tumor markers (CEA and Cyfra 21-1).

Abstract 2260: Classification of patients with lung cancer and benign lung disease via assessment of DNA methylation of SHOX2 and PTGER4 in plasma

Abstract Introduction: Annual screening for lung cancer (LC) with low-dose computed tomography (LDCT) is recommended in the US for specific high risk patients. Despite this, LC patients are frequently symptomatic at time of presentation and are often diagnosed with advanced stage disease associated with poor prognosis. At the same time the substantial rate of false positive findings of LDCT raised controversy about the risk profile of the groups eligible for LDCT screening. Non-invasive diagnostic tools may contribute to further risk stratification for screening candidates or patients with findings in LDCT. Aberrant DNA methylation detected in cell free-floating DNA has been shown to be clinically useful. Specifically, a DNA methylation panel of the genes SHOX2 and PTGER4 has been evaluated in three independent case-control studies comprising a total of 330 plasma specimens from LC patients and healthy individuals with promising results (AUC = 91 to 95%). Here, we report on further evaluation of the methylation marker panel in patients with malignant (LC) or benign lung disease (BLD). Material and Methods: For this case-control study 116 plasma specimen were collected from patients in Bulgaria and the US. The majority of patients were either diagnosed with LC (30 patients) or a BLD (50 patients). The remaining 36 subjects were healthy controls. A triplex real-time PCR assay for methylated DNA of SHOX2 and PTGER4 and the methylation independent ACTB control assay was developed. Total DNA was extracted from 3.5ml plasma samples and bisulfite converted utilizing a commercially available kit. After purification DNA was assayed in PCR triplicates. Cycle threshold values were aggregated utilizing a predefined algorithm. Receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC) were analysed. Results: The marker panel showed significant discriminatory power to distinguish LC cases from the remaining subjects (AUC = 0.84), including BLD patients (AUC = 0.83) and healthy subjects (AUC = 0.86). At a fixed specificity level of 95% the sensitivity for LC was 50%, while for a fixed 90% sensitivity the observed specificity was 53%. BLD and healthy subjects were similar with respect to the methylation markers (AUC = 0.55, p-value = 0.45). The clinically important group of COPD patients (18 cases) was significantly different from the LC group (AUC = 0.76, p-value &amp;lt; 0.01). Conclusions: Combination of DNA methylation markers SHOX2 and PTGER4 can be used to distinguish lung cancer patients from healthy subjects as well as patients with non-malignant diseases with high sensitivity at a reasonable false positive rate. The marker panel will be developed into a blood-based lung cancer diagnostic tool that may ultimately show clinical utility in combination with current imaging techniques to improve lung cancer risk stratification. Citation Format: Anne Schlegel, Oliver Hasinger, Selina Esche, Melanie Martini, Thomas König, Gunter Weiss. Classification of patients with lung cancer and benign lung disease via assessment of DNA methylation of SHOX2 and PTGER4 in plasma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2260.

Abstract 2247: A multicenter clinical trial of lung cancer circulating tumor cell assay with the largest sample size (1210 cases) in China

Abstract Background: As a novel biomarker of primary tumors, circulating tumor cells (CTCs) are well known to play an important role in cancer diagnosis, recurrence and metastasis, and disease monitoring. This study investigated the application of CTCs in lung cancer therapy by quantitative determination of folate receptor-positive CTCs. Methods: This study enrolled 1210 subjects (including 560 lung cancer patients, 350 patients with benign lung diseases, 150 healthy subjects and 150 non-lung cancer malignant tumor patients) and quantified the tiny amounts of CTCs in peripheral blood by negative enrichment using immunomagnetic beads in combination with folate receptor-directed polymerase chain reaction (PCR). Results: Following ROC curve analyses of CTC levels in the patients with benign lung diseases or benign lung diseases and the healthy subjects, the cut-off of CTCs was 8.70 CTC Units/3mL with a specificity of 88.2% (441/500) and a sensitivity of 79.6% (446/560) as determined by the method mentioned above. The ROC area under curve of more than 0.8 (AUC = 0.8797, P&amp;lt;0.0001) suggested the excellence of ligand-directed PCR to diagnose lung cancer. For stage II-IV lung cancer patients, the sensitivities were 70.8%(34/48), 79.6% (125/157), and 90.2% (185/205) respectively, and the sensitivity in stage I patients can reach up to 68.0% (102/150) as well. CTC levels appeared to have greater accuracy to diagnose lung cancer (AUC: 0.8833, P&amp;lt;0.0001) than the combination of the other four lung cancer-specific serum tumor markers (CEA, NSE, CYFRA21-1 and SCC)(AUC: 0.8557, P&amp;lt;0.0001). CTCs were related to tumor stage and tumor size other than pathological type. By quantitative determination of folate receptor-positive cells in 150 different types of cancers, the CTC level in lung cancer was found to be higher than those in gastric, breast, colorectal, liver, and esophageal cancers. Conclusions: LT-PCR achieves quantitative determination of folate receptor-positive CTCs to diagnose lung cancer with high sensitivity and specificity, and significantly greater diagnosis accuracy than the model of combined lung cancer-specific serum tumor markers. Citation Format: Jiatao Lou, Caicun Zhou, Jing Wu, Lihua Qiao, Xiaohui Liang, Xiaoqian Wang, Xiaoxia Chen, Xuefei Li, Chao Zhao. A multicenter clinical trial of lung cancer circulating tumor cell assay with the largest sample size (1210 cases) in China. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2247.

Video-assisted thoracoscopic surgery is a safe option for benign lung diseases requiring lobectomy

Lobectomy for benign lung disease is renowned to be technically complex and to be subjected to an increased complication rate. The objective of this study was to evaluate whether the results obtained with video-assisted surgery (VATS) in benign disease are comparable to those obtained in oncologic surgery, where VATS has been validated. We have reviewed the files of 246 consecutive patients who underwent VATS lobectomy from January 2012 to August 2015. The cohort was divided into two groups according to pathology (benign or malignant). Outcome parameters on scrutiny were demographics, pathology, duration of air leak, drainage and hospital stay, conversion, and perioperative complication rate. Comparisons were made with the χ 2 test and Student's t test; any p value ≤0.05 was considered as significant. Group 1 (36 patients) included patients who underwent lobectomy for benign disease and group 2 (210 patients) patients affected by lung cancer or pulmonary metastases. The two groups differed with reference to age (p<0.001), history of cancer (p<0.001), history of stroke (p=0.05), and the presence of pleural adhesions (p=0.03). There was no difference for duration of air leaks, chest tube drainage and hospital stay, conversion rate, and perioperative complication rate. We conclude that pathology did not impact on outcomes after VATS lobectomy. This study suggests that VATS is as a safe option in selected patients with benign disease requiring lobectomy, despite a more complex technical context.

Clinical value of serum trophoblast cell surface protein 2 (TROP2) antibody in non-small-cell lung cancer patients

Objective: This study was to explore the clinical role of serum trophoblast cell surface protein 2 (TROP2) antibody in patients with non-small-cell lung cancer (NSCLC).Materials and methods: We collected serum specimens from 117 NSCLC patients, 40 benign lung disease patients, and 60 healthy controls. TROP2 antibody concentrations were measured using enzyme-linked immunosorbent assay.Results: Serum TROP2 antibody levels were higher in the NSCLC group compared to the control group (p < 0.001). TROP2 antibody, at a cutoff value of 9.8 U/ml, showed good diagnostic performance for NSCLC.Conclusion: Measurement of TROP2 antibody might have diagnostic value for patients with NSCLC.