Identification of Potential microRNA Panels for Male Non-Small Cell Lung Cancer Identification Using Microarray Datasets and Bioinformatics Methods.

Background: Non-small cell lung cancer (NSCLC) is still one of the types of cancer with the highest death rates. MicroRNAs (miRNAs) play essential roles in NSCLC development. This study evaluates miRNA expression patterns and specific mechanisms in male patients with NSCLC. Methods: We report an integrated microarray analysis of miRNAs for eight matched samples of males with NSCLC compared to the study of public datasets of males with NSCLC from TCGA, followed by qRT-PCR validation. Results: For the TCGA dataset, we identified 385 overexpressed and 75 underexpressed miRNAs. Our cohort identified 54 overexpressed and 77 underexpressed miRNAs, considering a fold-change (FC) of ±1.5 and p < 0.05 as the cutoff value. The common miRNA signature consisted of eight overexpressed and nine underexpressed miRNAs. Validation was performed using qRT-PCR on the tissue samples for miR-183-3p and miR-34c-5p and on plasma samples for miR-34c-5p. We also created mRNA-miRNA regulatory networks to identify critical molecules, revealing NSCLC signaling pathways related to underexpressed and overexpressed transcripts. The genes targeted by these transcripts were correlated with overall survival. Conclusions: miRNAs and some of their target genes could play essential roles in investigating the mechanisms involved in NSCLC evolution and provide opportunities to identify potential therapeutic targets.

Next-Generation Sequencing in Lung Cancer Patients: A Comparative Approach in NSCLC and SCLC Mutational Landscapes.

Background: Lung cancer remains one of the most diagnosed malignancies, being the second most diagnosed cancer, while still being the leading cause of cancer-related deaths. Late diagnosis remains a problem, alongside the high mutational burden encountered in lung cancer. Methods: We assessed the genetic profile of cancer genes in lung cancer using The Cancer Genome Atlas (TCGA) datasets for mutations and validated the results in a separate cohort of 32 lung cancer patients using tumor tissue and whole blood samples for next-generation sequencing (NGS) experiments. Another separate cohort of 32 patients was analyzed to validate some of the molecular alterations depicted in the NGS experiment. Results: In the TCGA analysis, we identified the most commonly mutated genes in each lung cancer dataset, with differences among the three histotypes analyzed. NGS analysis revealed TP53, CSF1R, PIK3CA, FLT3, ERBB4, and KDR as being the genes most frequently mutated. We validated the c.1621A>C mutation in KIT. The correlation analysis indicated negative correlation between adenocarcinoma and altered PIK3CA (r = −0.50918; p = 0.0029). TCGA survival analysis indicated that NRAS and IDH2 (LUAD), STK11 and TP53 (LUSC), and T53 (SCLC) alterations are correlated with the survival of patients. Conclusions: The study revealed differences in the mutational landscape of lung cancer histotypes.

EBUS in optimizing non-small cell lung cancer diagnosis and treatment.

BACKGROUND: Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a commonly used minimally invasive method for the diagnosis and staging of lung cancer. In order to improve its diagnostic accuracy, rapid on-site cytologic evaluation (ROSE) is being utilized in some institutions. ROSE, performed by a cytopathologist in the examination room, allows the assessment of the adequacy of the collected samples, identifies malignant cells and sometimes establishes diagnosis on the spot, thus improving diagnostic sensitivity. As non-small cell lung carcinomas (NSCLC) require not only pathological subtyping, but also molecular characterization, obtaining the adequate amount of tissue is crucial. Only a limited number of studies have analyzed the suitability of EBUS-TBNA samples for assessment of epidermal growth factor receptor (EGFR), anaplastic lymphoma kinase (ALK) and programmed death-ligand 1 (PD-L1) status. AIM: We intended to examine the diagnostic yield of ROSE in NSCLC and the results and feasibility of molecular analysis performed on EBUS-TBNA small samples. METHODS: 100 patients with lung tumors and hilar and/or mediastinal lymphadenopathy on CT or PET/CT scans were retrospectively identified over a 3-year period, from a prospectively maintained EBUS-TBNA database. All examinations were accompanied by on-site cytological exam - ROSE, histopathological exam (HPE) and, in the case of NSCLC, molecular testing. After the sampling of the lymph nodes, specimens were Diff-Quik stained and a rapid preliminary diagnosis was established. Immunohistochemistry and mutational testing were performed using cell blocks. RESULTS: Adenocarcinoma was the most frequent diagnosis in both ROSE (34%) and histopathology (53%). Overall sensitivity and positive predictive value of ROSE in NSCLC, considering HPE the gold standard, were 92.18% and 93.65%, respectively, with a specificity and negative predictive value of 75% and 70.58%, respectively. All samples that were tested for EGFR mutation and ALK rearrangement were adequate for analysis. The adequacy ratio for PD-L1 was 91.66%; 37.5% of patients showed a high PD-L1 expression level, with a tumor proportion score TPS ≥50%. CONCLUSION: EBUS-TBNA is a valuable method for lung cancer diagnosis. ROSE proved to have a moderate prediction of the final diagnosis in NSCLC. Molecular analysis of EGFR, ALK and PD-L1 can be successfully accomplished on EBUS-TBNA small tissue samples.

Cryobiopsy in Lung Cancer Diagnosis-A Literature Review.

Optimizing the diagnosis of lung cancer represents a challenge, as well as a necessity, for improving the low survival of these patients. Flexible bronchoscopy with forceps biopsy is one of the key diagnostic procedures used for lung tumors. The small sample size and crush artifacts are several factors that can often limit access to a complete diagnosis, therefore leading to the need of repeating the bronchoscopy procedure or other invasive diagnostic methods. The bronchoscopic cryobiopsy is a recent technique that proved its utility in the diagnosis of both endobronchial and peripheral lung tumors. In comparison with conventional forceps biopsy, studies report a higher diagnostic yield and a superior quality of the collected samples for both the histopathological and the molecular diagnosis of lung cancer. This method shows promising results in sampling lung tissue, alone, or in conjunction with fluoroscopy or radial endobronchial ultrasound (r-EBUS). With a good safety and cost-benefit profile, this novel method has the potential to improve the diagnosis, and therefore the management of lung cancer patients. The objective of this narrative review is to provide a comprehensive review of the recent data regarding the advantages of cryobiopsy and r-EBUS in lung cancer diagnosis.

SERS-based differential diagnosis between multiple solid malignancies: breast, colorectal, lung, ovarian and oral cancer.

PURPOSE: Surface-enhanced Raman scattering (SERS) spectroscopy on serum and other biofluids for cancer diagnosis represents an emerging field, which has shown promising preliminary results in several types of malignancies. The purpose of this study was to demonstrate that SERS spectroscopy on serum can be employed for the differential diagnosis between five of the leading malignancies, ie, breast, colorectal, lung, ovarian and oral cancer. PATIENTS AND METHODS: Serum samples were acquired from healthy volunteers (n=39) and from patients diagnosed with breast (n=42), colorectal (n=109), lung (n=33), oral (n=17), and ovarian cancer (n=13), comprising n=253 samples in total. SERS spectra were acquired using a 532 nm laser line as excitation source, while the SERS substrates were represented by Ag nanoparticles synthesized by reduction with hydroxylamine. The classification accuracy yielded by SERS was assessed by principal component analysis-linear discriminant analysis (PCA-LDA). RESULTS: The sensitivity and specificity in discriminating between cancer patients and controls was 98% and 91%, respectively. Cancer samples were correctly assigned to their corresponding cancer types with an accuracy of 88% for oral cancer, 86% for colorectal cancer, 80% for ovarian cancer, 76% for breast cancer and 59% for lung cancer. CONCLUSION: SERS on serum represents a promising strategy of diagnosing cancer which can discriminate between cancer patients and controls, as well as between cancer types such as breast, colorectal, lung ovarian and oral cancer.

Soluble sPD-L1 and Serum Amyloid A1 as Potential Biomarkers for Lung Cancer.

BACKGROUND: The objective of this prospective study was to evaluate whether soluble programmed cell death-1/programmed cell death-ligand 1 (PD-1/PD-L1) and serum amyloid A1 (SAA1) are potential diagnostic, predictive or prognostic biomarkers in lung cancer. METHODS: Lung cancer patients (n=115) with advanced metastatic disease, 101 with non-small cell lung cancer, NSCLC (77 EGFR wild-type NSCLC patients on chemotherapy, 15 EGFR mutation positive adenocarcinoma patients, 9 patients with mPD-L1 Expression ≥50% NSCLC - responders to immunotherapy), and 14 patients with small cell lung cancer (SCLC) were examined. ELISA method was used to determine sPD-L1 and SAA1 concentrations in patients' plasma. RESULTS: Significantly higher blood concentrations of sPD-L1 and SAA1 were noted in lung cancer patients compared with a healthy control group. In PD-L1+ NSCLC patients, a significantly higher sPD-L1 level was noticed compared to any other lung cancer subgroup, as well as the highest average SAA1 value compared to other subgroups. CONCLUSIONS: It seems that sPD-1/PD-L1 might be a potential biomarker, prognostic and/ or predictive, particularly in patients treated with immunotherapy. Serum amyloid A1 has potential to act as a good predictor of patients' survival, as well as a biomarker of a more advanced disease, with possibly good capability to predict the course of disease measured at different time points.

Current Practice of Airway Stenting in the Adult Population in Europe: A Survey of the European Association of Bronchology and Interventional Pulmonology (EABIP).

BACKGROUND: Airway stenting (AS) commenced in Europe circa 1987 with the first placement of a dedicated silicone airway stent. Subsequently, over the last 3 decades, AS was spread throughout Europe, using different insertion techniques and different types of stents. OBJECTIVES: This study is an international survey conducted by the European Association of Bronchology and Interventional Pulmonology (EABIP) focusing on AS practice within 26 European countries. METHODS: A questionnaire was sent to all EABIP National Delegates in February 2015. National delegates were responsible for obtaining precise and objective data regarding the current AS practice in their country. The deadline for data collection was February 2016. RESULTS: France, Germany, and the UK are the 3 leading countries in terms of number of centres performing AS. These 3 nations represent the highest ranked nations within Europe in terms of gross national income. Overall, pulmonologists perform AS exclusively in 5 countries and predominately in 12. AS is performed almost exclusively in public hospitals. AS performed under general anaesthesia is the rule for the majority of institutions, and local anaesthesia is an alternative in 9 countries. Rigid bronchoscopy techniques are predominant in 20 countries. Amongst commercially available stents, both Dumon and Ultraflex are by far the most commonly deployed. Finally, 11 countries reported that AS is an economically viable activity, while 10 claimed that it is not. CONCLUSION: This EABIP survey demonstrates that there is significant heterogeneity in AS practice within Europe. Therapeutic bronchoscopy training and economic issues/reimbursement for procedures are likely to be the primary reasons explaining these findings.

Endobronchial ultrasound - one year of experience in clinical practice.

BACKGROUND AND AIM: Endobronchial ultrasound (EBUS) is a recent minimally invasive, safe examination method for the mediastinum, with a good diagnostic precision. This method makes possible real time examination with transbronchial fine needle aspiration, diagnostic transbronchial needle aspiration (TBNA) and staging of non-small pulmonary tumors, as well as diagnosis of mediastinal and hilar adenopathies of various causes. METHODS: We present the experience of the Bronchoscopy Department of the Pulmonology Clinic of Cluj-Napoca with EBUS-TBNA as a tool for the diagnosis and staging of tumors in contact with the bronchial wall and mediastinal and hilar adenopathies of unknown etiology. During the period August 2014 - January 2016 we examined 152 patients with no direct or indirect signs of lung tumor in traditional bronchoscopy. Rapid on site evaluation (ROSE) was available for all patients. RESULTS: Our study is a retrospective study of 152 EBUS-TBNA examinations. The average age of our patients was 54.43 years and 64% came from urban and 36% from rural background. EBUS-TBNA brought the final histological confirmation (tumors, sarcoidosis, limphoma) in 82.8% of the cases. A tumor confirmation was obtained in 95% of the patients who were suspected of having tumor. For a better understanding of the importance of this method in the daily clinical practice we present a case of peripheral pulmonary neoplasm with mediastinal and hilar adenopathies, where the contribution of EBUS-TBNA to a rapid diagnosis was essential. CONCLUSION: By the introduction of this method in our country one year ago, we can diagnose patients with lung and mediastinal tumors, which cannot be diagnosed by traditional bronchoscopy. This brings a valuable contribution to the improvement of lung cancer staging and diagnostic.

The use of EBUS-TBNA and ROSE in the diagnosis of lung cancer.

Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a minimally invasive method for diagnosing and staging of lung cancer. EBUS-TBNA obtained small specimens. Rapid on-site examination (ROSE) is a rapid, real-time examination method. The aim of our study is to evaluate the impact of ROSE on adequate specimen sampling, rapid results and high diagnostic rate. We present the experience of the Department of Bronchology, "Leon Daniello" Clinic of Pulmonology, Cluj-Napoca, Romania, with EBUS-TBNA as a tool for diagnostic of adenopathies of unknown etiology. We evaluated the diagnostic capacity of ROSE for malignant tumors, by considering the histopathological examination as the diagnostic "gold standard". In our retrospective and descriptive study, we analyze the data of 147 EBUS-TBNA examinations with ROSE and histopathological exam, performed for diagnostic purposes for hilar and mediastinal adenopathies of unknown origin. The age of the patients varied from 21 to 80 years, with an average age of 54.36 years. There were 98 male patients, representing 66.66% of the group. From the total of 90 cases of malignancy, 72 (80%) cases were identified as a primary lung tumor, 13 (14.44%) cases were identified as lymphoma, and five cases as malignant tumor of extrapulmonary origin. The sensitivity of the ROSE is 85.71%. By the introduction of this method, EBUS-TBNA with ROSE, in our country, we can diagnose patients with lung and mediastinal tumors, which cannot be diagnosed by traditional bronchoscopy. This brings a valuable contribution to the improvement of lung cancer diagnostic.

Endobronchial ultrasound EBUS--a new method for the diagnosis and staging of lung cancer.

In this paper we present a new method, endobronchial ultrasound (EBUS), which appeared recently among the tools of the pulmonologist for the diagnosis and staging of lung cancer. Endobronchial ultrasound transbronchial needle aspiration (EBUS-TBNA) provides the opportunity for obtaining tissue samples required for the histologic and cytologic diagnosis of lung cancer. The advantages of EBUS have to be made popular as it is a minimally invasive method, safe, simple, fast, also with a superior cost/benefit ratio compared to any previously used methods.

Mounier-Kuhn syndrome.

Mounier-Kuhn syndrome or congenital tracheobronchomegaly is a rare clinical and radiologic condition. It is characterized by a tracheal and bronchial dilation. Fewer than 100 cases have been reported in the medical literature since the original description in 1932. The first utilization of bronchoscopy for diagnosis of this condition was recorded by Lemoine only in 1949. The cause of this condition is not clearly understood; however, histopathologic findings of atrophy of smooth muscles and elastic tissue in the trachea and main bronchi have been observed. Tracheobronchomegaly can be associated with tracheal and bronchial diverticuli.

[Esophageal pathology with respiratory impact]. / Patologie esofagiana cu rasunet asupra aparatului respirator.

We present some of digestive diseases: achalazia, esophageal diverticula and esophageal fistula, which in their evolution determine respiratory symptoms: dispnea, cough and imitate until confusion respiratory diseases. The diagnosis is most of the time difficult and it needs special investigations. A better knowledge of these diseases helps us to avoid mistakes in diagnosis.

[Update in bronchoscopic techniques]. / Update tehnici bronhoscopice.

Bronchoscopy is a central technique in diagnosing lung cancer, but also in different therapeutic approaches. The most common indication for bronchoscopy is for tissue sampling and determining the extent of lung cancer. Established diagnostic techniques are forceps biopsy, aspiration or brush cytology sampling, or needle aspiration. Laser therapy, electrocautery, cryotherapy and stenting are well-described techniques for the palliation of symptoms due to airway involvement in patients with advanced stages. Newer technologies, with an established role in clinical practice, are endobronchial ultrasound, autofluorescence bronchoscopy, and electromagnetic navigation. Other technologies, such as magnification, narrow-band imaging and confocal fluorescence microendoscopy, are in development for the use within the airways.

[Imaging in screening and diagnosis of lung cancer]. / Imagistica în screening-ul si diagnosticul cancerului bronho-pulmonar.

While the lung cancer is detected, the majority of the patients could not be surgical cured. Thus appears the great necessity of prevention, of a more efficient treatment and the developing of new methods for early detection. The first screening trials for lung cancer had negative results. None of the studies had no prove in decreasing of mortality by screening for lung cancer. The lack of sensitivity of thoracic radiographs and sputum cytology was leading the investigators to use the monoclonal antibodies in detecting the neoplastic changes in sputum, the spiral computerized tomography with low dose of radiations, the autofluorescence bronchoscopy and more recent the positron emission tomography, offering the possibility of reducing mortality by early detection in lung cancer.