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Background. MYC (v-myc avian myelocytomatosis viral oncogene homolog) is one of the most frequently amplified genes in lung tumors. For the analysis of gene copy number variations, dPCR (digital PCR) is an appropriate tool. The aim of our study was the assessment of dPCR for the detection of MYC copy number variations (CNV) in lung tissue considering clinicopathological parameters. Material and Methods. MYC status was analyzed with dPCR as well as qPCR (quantitative PCR) using gDNA (genomic DNA) from tumor and adjacent nontumor tissue samples of lung cancer patients. The performance of MYC was estimated based on the AUC (area under curve). Results. The results of the MYC amplification correlated significantly between dPCR and qPCR (rS=0.81, P<0.0001). The MYC copy number revealed by dPCR showed statistically significant differences between tumor and adjacent nontumor tissues. For discrimination, a sensitivity of 43% and a specificity of 99% were calculated, representing 55 true-positive and one false-positive tests. No statistically significant differences could be observed for age, sex, and smoking status or the clinicopathological parameters (histological subtype, grade, and stage). Conclusion. The results of the study show that dPCR is an accurate and reliable method for the determination of MYC copy numbers. The application is characterized by high specificity and moderate sensitivity. MYC amplification is a common event in lung cancer patients, and it is indicated that the determination of the MYC status might be useful in clinical diagnostics.

Objectives MET c-Cbl binding site mutations constitute about 2 % of MET exon 14 alterations in lung cancer. Preclinical data suggests regarding these mutations as functional analogs of MET exon 14 skipping mutations, but clinical validation is lacking. Results We report the case of a patient with metastastic lung adenocarcinoma harboring a c-Cbl binding site alteration and demonstrate clinical, radiological and metabolic response to crizotinib with a PFS of 10.6 months. As escape mechanism, a typical MET resistance mutation could be identified. Conclusion MET c-Cbl binding site mutations should be regarded as a distinct subtype of MET exon 14 alterations. Patients with lung cancer harboring such mutations should be offered targeted therapy.

Endobronchial ultrasound-guided transbronchial needle aspirations (EBUS-TBNAs) are well established for staging lung cancer. A growing number of publications report on lymphoma diagnosis via EBUS-TBNA-acquired cytology; however current guidelines recommend histologic diagnosis. Research on the value of EBUS-TBNA-acquired cytology versus surgical-acquired histology in the diagnosis of lymphoma is lacking.We conducted a retrospective review of patients with mediastinal lymphoma diagnosed between 2010 and 2016. Mediastinal lymphadenopathy was accessible through both EBUS-TBNAs and surgical procedures. All data were extracted from our clinic's medical database and analyzed.Fifty-one patients newly diagnosed with lymphoma in the mediastinum were identified (median age, 43.5 years; mean age, 48.6 ± 20.6 years). A minimally invasive procedure was performed as a first diagnostic step in 29 patients, whereas surgical biopsy was performed in the remaining 22. The time to final diagnosis was significantly longer if a minimally invasive procedure was performed first compared with a surgical procedure (mean, 44 days [median, 38 days] vs 16 days [median, 8 days]; p0.030). The number of procedures to obtain a final diagnosis ranged from one to five (median, 2 procedures per patient) in the EBUS-TBNA group. This was significantly higher than that in the surgical group (median, 1 procedure per patient; p0.00005).We demonstrate that surgical biopsies are safe and well tolerated for lymphoproliferative disease diagnosis and lead to a final diagnosis in the shortest possible time. Unnecessary procedures were significantly reduced if a surgical biopsy was performed as the first step.

Major advances in molecular profiling for available targeted treatments and immunotherapy for lung cancer have significantly increased the complexity of tissue-based diagnostics. Endobronchial ultrasound-guided transbronchial needle aspirations (EBUS-TBNA) are commonly performed for diagnostic biopsies and lymph node staging. EBUS-TBNA has increasingly become one of the main sources of tumor cells for molecular analyses. As a result, there is a growing need for high quality EBUS-TBNA samples with adequate cellularity. This has increased the technical demands of the procedure and has created additional challenges, many of which are not addressed in the current EBUS guidelines. This review provides an overview of current evidence on the technical aspects of EBUS-TBNA in light of comprehensive sample processing for personalized lung cancer management. These include sonographic lymph node characterization, optimal needle choice, suction biopsy technique, and the role of rapid on-site evaluation. Attention to these technical details will be important to maximize the throughput of EBUS-TBNA biopsies for molecular testing.

Objective BRAF mutations have been subtyped in three functional classes with different oncogenic modes of action. The clinical impact of BRAF mutational subtypes in non–small-cell lung cancer (NSCLC) remains to be defined. So far, ambiguous results were reported from analyses of heterogeneous patient cohorts. Methods We studied patients with metastatic or recurrent NSCLC who were sequentially enrolled in precision oncology programs at two large German lung cancer centres from 2009 to 2019. The study period allowed evaluating the specific impact of BRAF V600E-targeting. Results In a cohort of 72 patients, BRAF mutation subtyping revealed p.V600E mutations in 31 cases (43%), whereas 41 cases (57%) harboured 18 different BRAF mutational subtypes of functional classes II/III. Functionally relevant comutations were observed in 6.4% of class I, and 24.4% of class II/III BRAF mutations. Most patients were treated with chemotherapy. Targeted therapy was administered in 11 patients with a response rate of 72.7%. PD-1/PD-L1-immunotherapy was given in 14 patients with a response rate of 28.6%. Overall survival of patients with BRAF-mutated NSCLC was inferior (HR 1.38, p = 0.048) as compared to patients with BRAF wild-type cancers. Median time-to-treatment-failure with BRAF-targeting agents was shorter as compared to approved targeted therapy of other oncogenic drivers (HR 1.97, p = 0.05). Survival outcomes were not impacted by BRAF mutation subtype functional class. Conclusions Patients with BRAF-mutated NSCLC have an inferior prognosis, which is not determined by BRAF mutation functional class. In contrast to NSCLC with other tractable driver mutations, BRAF-mutated NSCLC exhibit high susceptibility to immune checkpoint inhibitors.

Background/introduction In contrast to patients who present with advanced stage lung cancer and associated poor prognosis, patients with early-stage lung cancer may be candidates for curative treatments. The results of the NELSON lung cancer screening trial are expected to stimulate the development and implementation of a lung cancer screening strategy in most countries. Widespread use of chest computed tomography scans will also result in the detection of solitary pulmonary nodules. Because reliable biomarkers to distinguish between malignant and benign lesions are lacking, tissue-based histopathological diagnostics remain the gold standard. In this study, we aimed to establish a test to assess the predictive ability of DNA hypermethylation of SHOX2 and PTGER4 in plasma to discriminate between patients with 1.) lung cancer, 2.) benign lesions, and 3.) patients with chronic obstructive pulmonary disease (COPD). Patients and methods We retrospectively analysed SHOX2 and PTGER4 methylation in 121 prospectively collected plasma samples of patients with lung cancer (group 1A), benign lesions (group 1B), and COPD without nodules (group 2). Results PTGER4 DNA hypermethylation was more frequently observed in patients with lung cancer than in controls (p = 0.0004). Results remained significant after correction for tumour volume, smoking status, age, and eligibility for the NELSON trial. Conclusions Detection of methylated PTGER4 in plasma DNA may serve as a biomarker to support clinical decision-making in patients with pulmonary lesions at lung cancer screening in high-risk populations. Further exploration in prospective studies is warranted.

Introduction: In the era of targeted therapy of lung cancer, availability of adequate specimen samples for molecular analysis is increasingly important. EBUS-TBNA is often used for tissue acquisition, and the choice of the needle might influence the rate of successful molecular analysis. This prospective study evaluated quality and quantity of specimens obtained using different EBUS-TBNA needles. Methods: Consecutive patients with advanced lung cancer referred for EBUS-TBNA were included. A 22G steel needle (N1), a more flexible needle with a nitinol stylus (N2), or a core biopsy needle (N3) were used for EBUS-TBNA. Specimens were placed on a slide and weighed before transfer of tissue into formalin and sending for histopathological analysis, immunohistochemistry and next-generation sequencing (NGS). Tumour cellularity was assessed by an experienced pathologist. Results: 50 patients (28 male;mean 62.9y) with NSCLC (adeno n=38; squamous cell n=5; large cell n=3; other n=4) were included in the analysis. 25, 12 and 13 specimens were obtained with N1,N2 and N3, respectively. Mean specimen weight was 42.6±38.0 mg. Tumour cellularity was 1000 tumour cells in 6, 5, 6, 3, 8, 9 and 12 cases, respectively. Immunohistochemistry could be performed in all but 3 cases (2 with N2, 1 with N3). NGS could be done in 41 cases (82.0%; failure in 2 cases with N1, 6 with N2, 1 with N3). Tumour cellularity, sample weight and rate of successful NGS were significantly lower when EBUS-TBNA was performed with N2. Conclusion: The needle used for EBUS-TBNA had an impact on quantity and quality of tissue obtained for molecular analysis in NSCLC.

The detection of molecular alterations is crucial for the individualized treatment of advanced non-small cell lung cancer (NSCLC). Missing targetable alterations may have a major impact on patient’s progression free and overall survival. Although laboratory testing for molecular alterations has continued to improve; little is known about how biopsy technique affects the detection rate of different mutations. In the retrospective study detection rate of epidermal growth factor (EGFR) mutations in tissue extracted by bronchoscopic cryobiopsy (CB was significantly higher compared to other standard biopsy techniques. This prospective, randomized, multicenter, single blinded study evaluates the accuracy of molecular genetic characterization of NSCLC for different cell sampling techniques. Key inclusion criteria are suspected lung cancer or the suspected relapse of known NSCLC that is bronchoscopically visible. Patients will be randomized, either to have a CB or a bronchoscopic forceps biopsy (FB). If indicated, a transbronchial needle aspiration (TBNA) of suspect lymph nodes will be performed. Blood liquid biopsy will be taken before tissue biopsy. The primary endpoint is the detection rate of molecular genetic alterations in NSCLC, using CB and FB. Secondary endpoints are differences in the combined detection of molecular genetic alterations between FB and CB, TBNA and liquid biopsy. This trial plans to recruit 540 patients, with 178 evaluable patients per study cohort. A histopathological and molecular genetic evaluation will be performed by the affiliated pathology departments of the national network for genomic medicine in lung cancer (nNGM), Germany. We will compare the diagnostic value of solid tumor tissue, lymph node cells and liquid biopsy for the molecular genetic characterization of NSCLC. This reflects a real world clinical setting, with potential direct impact on both treatment and survival.

Background The optimal treatment for patients with locally advanced non-small-cell lung-cancer (NSCLC) cT4 cN0/1 cM0 is still under debate. The purpose of this study was to examine the long-term survival of cT4 cN0/1 cM0 NSCLC patients undergoing induction chemotherapy and concurrent radiochemotherapy before surgery. Methods All consecutive patients with confirmed NSCLC (cT4 cN0/1 cM0) treated with neoadjuvant chemotherapy, concurrent radiochemotherapy (RT/CTx) (45–46 Gy) and surgical resection between 2000 and 2015 were included. According to the UICC guidelines (8th edition), T4 stage was reanalysed by an expert radiologist. The mediastinal staging was performed by systematic EBUS-TBNA or mediastinoscopy. The primary end-point was overall-survival (OS). The power to detect an increase of early tumour-associated mortality (hazard ratio > 3.5) within the first 5 years after treatment in comparison to late deaths beyond 96 months was >80%. Results Overall, 67 patients were treated with concurrent RT/CTx. T4 criteria were fulfilled by all patients, and multiple T4 criteria by 53 patients. Seventy percent of patients had an initial PET/CT staging. The median follow-up period was 134 months. OS rates at 2, 5, 10 and 15 years were 83.6 ± 4.5%, 65.4 ± 5.9%, 53.3 ± 6.3% and 36.6 ± 6.8%, respectively. A total of 44.8% of patients achieved a pathologic complete response. In multivariable analysis, ypT category was the most predictive factor. OS at 5 years for ypT0 (n = 31) was 80.5%, and ypT1 (n = 11) was 62.5%. Main sites of failure were brain and pulmonary metastases in seven and three patients, respectively. The intercurrent annual death rate was estimated from the survival curve beyond 96 months and was found to be 4.75% (95% CI 2.40–9.27%). No significant increased mortality was observed during the first 5 years (annual death rate: 8.31% [95% CI 5.60–12.24%], hazard-ratio = 1.72 [95% CI 0.81–3.65]). Conclusions The effectiveness of this trimodality schedule is high in patients with cT4 cN0/1 cM0 NSCLC with excellent local control rates. Considering the annual death rate beyond 8 years of survival as an intercurrent death rate due to comorbidity, this treatment schedule reduces annual mortality to background even in the first 5 years after therapy.

Background: Endobronchial ultrasound (EBUS) with transbronchial needle aspiration increases the diagnostic yield of lung cancer staging. The left adrenal gland (LAG) is a common site for lung cancer metastasis. The modality of transesophageal examination with an EBUS bronchoscope (EUS-B) routinely for LAG has not been assessed. Objective: The aim of this study was to prospectively assess if evaluation and tissue sampling of the LAG could routinely be implemented in an EBUS procedure. Methods: Patients referred for EBUS between March and August 2017 had assessment of the LAG via EUS-B. Fine-needle aspiration (FNA) was performed in cases with a suspicious LAG. The detection rate, procedure time, and learning curve of four experienced EBUS-bronchoscopists was assessed, plus the diagnostic accuracy and complication rate of FNA. Results: In total, 313 consecutive patients were included. The overall LAG detection rate was 87.5%. After the initial learning curve, the detection rate for all four bronchoscopists was >93%. The detection rate did not correlate with any patient characteristics. EUS-B-FNA revealed nine LAG metastases, with a sensitivity, specificity, and accuracy of 75%, 100%, and 99%, respectively. The mean EUS-B operation time was 194.4 s, with 594.8 s for FNA. There were no FNA-associated complications. Conclusions: Evaluation of the LAG with EUS-B could routinely be included in an EBUS procedure if necessary. A high detection rate can be achieved after an initial learning period. FNA of the LAG was feasible and safe. EUS-B of the LAG could be integrated into the usual EBUS/EUS-B procedure in lung cancer staging workup.

Objective Current predictive biomarkers for PD-1 (programmed cell death protein 1)/PD-L1 (programmed death-ligand 1)-directed immunotherapy in non-small cell lung cancer (NSCLC) mostly focus on features of tumour cells. However, the tumour microenvironment and immune context are expected to play major roles in governing therapy response. Against this background, we set out to apply context-sensitive feature selection and machine learning approaches on expression profiles of immune-related genes in diagnostic biopsies of patients with stage IV NSCLC. Methods RNA expression levels were determined using the NanoString nCounter platform in formalin-fixed paraffin-embedded tumour biopsies obtained during the diagnostic workup of stage IV NSCLC from two thoracic oncology centres. A 770-gene panel covering immune-related genes and control genes was used. We applied supervised machine learning methods for feature selection and generation of predictive models. Results Feature selection and model creation were based on a training cohort of 55 patients with recurrent NSCLC treated with PD-1/PD-L1 antibody therapy. Resulting models identified patients with superior outcomes to immunotherapy, as validated in two subsequently recruited, separate patient cohorts (n = 67, hazard ratio = 0.46, p = 0.035). The predictive information obtained from these models was orthogonal to PD-L1 expression as per immunohistochemistry: Selecting by PD-L1 positivity at immunohistochemistry plus model prediction identified patients with highly favourable outcomes. Independence of PD-L1 positivity and model predictions were confirmed in multivariate analysis. Visualisation of the models revealed the predictive superiority of the entire 7-gene context over any single gene. Conclusion Using context-sensitive assays and bioinformatics capturing the tumour immune context allows precise prediction of response to PD-1/PD-L1-directed immunotherapy in NSCLC.

Background Deregulated signal transduction pathways play a key role in development, progression and therapeutic resistance of non-small cell lung cancers (NSCLC). The purpose of this study is to assess the downstream markers of two well-characterized pathways and to correlate them with clinical outcome. Design 670 patients with metastatic NSCLC were prospectively enrolled in a comprehensive biomarker profiling program at a single center from 2012 to 2016. Phosphorylation of extracellular signal-regulated kinase (p-ERK), and protein kinase B (p-AKT) was assessed by standardized immunohistochemistry. Product of scores for quantity and quality of staining were calculated (immunoreactive score, 0–9). Somatic mutations of Kirsten rat sarcoma viral oncogene homolog [KRAS], epithelial growth factor receptor [EGFR], v-Raf murine sarcoma viral oncogene homolog B [BRAF] and phosphatidylinositol 3-kinase [PIK3CA]) were detected by Sanger (2012-03/2015) and amplicon NGS (04/2015-02/2016). Patients enrolled during the first year (2012) were used as discovery cohort. Patients enrolled from 2013 to 02/2016 were used as validation cohort. Clinical data were retrieved from the electronic medical records and were analyzed retrospectively. Results Using a discovery cohort, we identified an immunoreactive score of p-ERK ≥3 to be prognostically relevant. The validation cohort confirmed that higher levels of p-ERK correlated with worse overall survival (OS) and higher proportion of RAS mutations. Multivariate analysis including established risk factors such EGFR, ALK or ROS mutations and metastatic disease showed a trend of a detrimental effect of high p-ERK on OS (HR 1.23, CI 0.94–1.59, p = 0.131 for p-ERK immunoreactive score ≥3) and time to treatment failure after first-line therapy in the validation cohort. Phosphorylated AKT did not correlate with clinical outcome. Conclusion While serving as a prognosticator in univariate analysis, highly phosphorylated ERK does not convey a significant prognostic effect for OS in the presence of other prognostic factors. Phosphorylated ERK indicates a higher activity of RAS in advanced NSCLC.

Background Heart exposure to ionizing irradiation can cause ischaemic heart disease. The partial heart volume receiving ≥5 Gy (heartV5) was supposed to be an independent prognostic factor for survival after radiochemotherapy for locally advanced non-small-cell lung cancer (NSCLC). But validation of the latter hypothesis is needed under the concurrent risks of lung cancer patients. Patients and methods The ESPATUE phase III trial recruited patients with potentially operable IIIA(N2)/selected IIIB NSCLC between 01/2004 and 01/2013. Cisplatin/paclitaxel induction chemotherapy was given followed by neoadjuvant radiochemotherapy (RT/CT) to 45 Gy (1.5 Gy bid/concurrent cisplatin/vinorelbine). Operable patients were randomized to definitive RT/CT(arm A) or surgery (arm B) and therefore were treated at two different total dose levels of radiotherapy. HeartV5 and mean heart dose (MHD) were obtained from the 3D radiotherapy plans, the prognostic value was analysed using multivariable proportional hazard analysis. Results A total of 161 patients were randomized in ESPATUE, heartV5 and MHD were obtained from the 3D radiotherapy plans for 155 of these [male/female:105/50, median age 58 (33–74) years, stage IIIA/IIIB: 54/101]. Power analysis revealed a power of 80% of this dataset to detect a prognostic value of heartV5 of the size found in RTOG 0617. Multivariable analysis did not identify heartV5 as an independent prognostic factor for survival adjusting for tumour and clinical characteristics with [hazard ratio 1.005 (0.995–1.015), P=0.30] or without lower lobe tumour location [hazard ratio 0.999 (0.986–1.012), P=0.83]. There was no influence of heartV5 on death without tumour progression. Tumour progression, and pneumonia were the leading causes of death representing 65% and 14% of the observed deaths. Conclusions HeartV5 could not be validated as an independent prognostic factor for survival after neoadjuvant or definitive conformal radiochemotherapy. Tumour progression was the predominant cause of death. Register No. Z5 - 22461/2 - 2002-017 (German Federal Office for Radiation Protection).

OA embargo Objectives Chromosomal rearrangements involving ROS1 define a rare entity of lung adenocarcinomas with exquisite sensitivity to molecularly targeted therapy. We report clinical outcomes and genomic findings of patients with ROS1-positive lung cancer who were prospectively identified within a multiplex biomarker profiling program at the West German Cancer Center. Methods Standardized immunohistochemical (IHC) analysis, fluorescence in situ hybridization (FISH), and hotspot mutation analyses were performed in 1345 patients with advanced cancer, including 805 patients with metastatic lung adenocarcinoma. Clinical and epidemiological data were retrieved from the institutional database. Results ROS1 positivity by IHC analysis was detected in 25 patients with lung cancer (4.8% of lung adenocarcinomas), including 13 patients (2.5%) with ROS1 FISH positivity with a cutoff of at least 15% of events. Of the ROS1 IHC analysis–positive cases, 36% presented with concomitant oncogenic driver mutations involving EGFR (six cases, five of which were clinically validated by response to EGFR-targeting agents), KRAS (two cases), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha gene (PIK3CA), and BRAF. Three cases initially classified as ROS1 FISH–negative passed the threshold of 15% positive events when repeat biopsies were analyzed at progression. The median overall survival of the ROS1-positive patients (104 months) was significantly superior to that of the 261 patients with EGFR/anaplastic lymphoma kinase/ROS1–negative lung adenocarcinoma (24.4 months, p = 0.044). Interestingly, the overall survival of the 13 ROS1-positive patients with lung cancer from initiation of pemetrexed-based chemotherapy was significantly prolonged when compared with that of 169 pemetrexed-treated patients with EGFR/anaplastic lymphoma kinase/ROS1–negative adenocarcinoma (p = 0.01). Conclusions ROS1-positive metastatic lung adenocarcinomas frequently harbor concomitant oncogenic driver mutations. Levels of ROS1 FISH–positive events are variable over time. This heterogeneity provides additional therapeutic options if discovered by multiplex biomarker testing and repeat biopsies. © 2016 International Association for the Study of Lung Cancer

Background Lung cancer is the major cause of cancer death worldwide. Mediastinal lymph node staging is important for pretreatment lung cancer management. Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a well established method for mediastinal lymph node staging. Although EBUS-TBNA samples are much smaller than surgical lymph node biopsies, histopathological evaluation and molecular testing can successfully be performed. One step nucleic acid amplification (OSNA), which measures cytokeratin 19 (CK19) mRNA concentration, is a target marker that is gaining importance in quick detection of lymph node metastases in breast cancer and other cancers. Recent publications suggest accurate and rapid detection of lung cancer metastases in surgically removed lymph nodes. In this study we aimed to investigate if CK19 mRNA detection via OSNA is feasible to accurately detect lymph node metastases in lung cancer patients using EBUS-TBNA samples. Materials and Methods A total of 102 EBUS-TBNA samples of 55 patients were collected. EBUS-TBNA was performed in lymph nodes exceeding 5 mm. OSNA was performed using a ready to use amplification kit (Lynoamp; Sysmex, Kobe, Japan) in the RD-100 I, an automated real-time detection system (Sysmex). Results Histopathological analysis confirmed malignancy in 90 cases and excluded malignancy in 12 cases. Overall sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 78.9%, 58.3%, 93.4%, 26.9%, and 76.5%, respectively. Conclusions One step nucleic acid amplification is feasible for EBUS-TBNA lymph node samples of lung cancer patients, but CK19 mRNA is an inaccurate marker, which is unlikely to be useful as an adjuvant test for EBUS-TBNA. Further studies are required to define the optimal sample size and sampling method.

BACKGROUND: Lung cancer is the leading cause of death for male and female cancer patients alike. Early diagnosis improves prognosis. A blood test would be a valuable support. OBJECTIVE: Infrared spectroscopy provides a label-free biochemical fingerprint of a sample. A study was conducted on 161 patients with initial cancer suspicion to identify and verify spectral biomarker candidate patterns to detect non-small cell lung carcinoma (NSCLC). METHODS: Blood serum and plasma samples were analysed with an automated FTIR spectroscopic system. Two pattern recognition algorithms and two classifiers were applied. Monte Carlo cross validation was performed with linear discriminant analysis and random forest classification. RESULTS: Marker patterns for the discrimination of cancer from clinically relevant disease control patients were identified in FTIR spectra of blood samples. An accuracy of up to 79% was achieved. Squamous cell and adenocarcinoma patients were separable with an accuracy of 80%. CONCLUSIONS: The study demonstrates the applicability of FTIR spectroscopic blood testing for lung cancer detection. Evidence for cancer subtype discrimination is given. With an improved performance, the method could be developed as a routine diagnostic tool for blood testing detecting NSCLC.

Until few years ago non-specific cytotoxic agents were considered the tip of the arrow as first line treatment for lung cancer. However; age > 75 was considered a major drawback for this kind of therapy. Few exceptions were made by doctors based on the performance status of the patient. The side effects of these agents are still severe for several patients. In the recent years further investigation of the cancer genome has led to targeted therapies. There have been numerous publications regarding novel agents such as; erlotinib, gefitinib and afatinib. In specific populations these agents have demonstrated higher efficiency and this observation is explained by the overexpression of the EGFR pathway in these populations. We suggest that TKIs should administered in the elderly, and with the word elderly we propose the age of 75. The treating medical doctor has to evaluate the performance status of a patient and decide the best treatment in several cases indifferent of the age. TKIs in most studies presented safety and efficiency and of course dose modification should be made when necessary. Comorbidities should be considered in any case especially in this group of patients and the treating physician should act accordingly. OA gold - CA extern

Background: EBUS guided trans-bronchial biopsy became routine in diagnosis of peripheral pulmonary lesions (PPL). Suction catheter-biopsy is a technique for obtaining a tissue sample from peripheral lung parenchyma. Aim of this study was to evaluate diagnostic efficiency, feasibility and safety of EBUS guided suction catheter-biopsy (SCB) in comparison to trans-bronchial biopsy (TBB) in diagnosis of PPL. The main intention was to demonstrate non-inferiority of the technique over trans-bronchial biopsy, especially when used under navigation of the EBUS. Methods: Radial EBUS probe (UM-3R, Olympus Co, Japan.) without guiding sheath was used to navigate suction catheter and TBB forceps to the PPL. The catheter was connected to the collection canister via vacuum pump. The SCB specimens were fixed with 10% buffered formalin. Results: There were 168 patients enrolled in this study; 69.9% males and 30.1% females. Main lesion diameter was 4.1±1.9 cm. Majority of patients, 131(77.9%) were diagnosed with lung cancer. Per-biopsy calculated sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for EBUS-SCB were 92.4%, 100%, 100% and 67.7%, respectively. Corresponding values for EBUS-TBB were 92.3%, 100%, 100% and 69.7%. Only the size of the lesion significantly influenced (p=0.005) diagnostic performance. Complications occurred in 2 patients; one pneumothorax and one excessive bleeding. Conclusion: EBUS guided SCB is efficient, feasible and safe in diagnosis of peripheral lung cancer. The technique is complementary to trans-bronchial biopsy. OA gold - CA extern

Mutated RAS onco-proteins are key drivers across many cancers. The distribution of somatic RAS mutations varies between cancer entities. Retrospective analyses have associated some RAS mutations with distinct clinical outcomes. However, the clinical impact of the full spectrum of RAS mutations in their disease contextuality remains to be defined. To improve upon this situation, we studied genomically and clinically annotated, prospectively recruited cohorts of patients with RAS-mutated metastatic lung cancer and colorectal cancer. Mutational spectra were compared with predictions derived from analyzing the mutagenic impact at the genome level for each entity. Interestingly, we found concordance of predicted signatures with those actually observed in our patients. Thus, composition of the functionally active RAS mutational subtypes is primarily determined by the mutagenic context. Most RAS mutations seemed dominant oncogenic drivers with entity-dependent clinical outcomes. RAS comutations were enriched in tumors harboring class 2/3 BRAF mutations, highlighting the functional dependency of some mutated BRAF isoforms on RAS. With our dataset, we established a probabilistic model for cross-entity comparison of the prognostic impact of specific RAS mutational subtypes. The resulting prognostic clusters showed largely consistent clinical categorizations in both entities. This suggests mutant subtype-specific functional properties leading to similar clinical effects. A notable exception is KRAS G12C, which imparted an adverse prognosis only in colorectal cancer. Our findings provide a framework for risk stratification of specific RAS mutations across several cancer entities, which is required to guide the analysis of clinical findings in patients treated with direct RAS inhibitors or agents targeting downstream pathways.

ZusammenfassungRebiopsien bei primären Lungenkarzinomen sowie deren Metastasen werden im klinischen Alltag immer wichtiger, da sich die Therapien weiterentwickeln und häufig gezielte weitere Behandlungsstrategien erst nach immunhistochemischen oder/und molekularen Veränderungen zugelassen sind. Prinzipiell können fast alle Rezidive bzw. progrediente Tumore biopsiert werden, allerdings häufig nur mittels invasiver Maßnahmen. Hier muss der Aufwand und das Risiko des Patienten mit dem zu erwartenden Benefit durch die Rebiopsie in jedem einzelnen Fall überlegt und vorher mit dem Patienten besprochen werden. In der Übersichtsarbeit werden die Indikationen bei Rezidiv und progredienter Erkrankung sowie die Risiken diskutiert und Alternativen zur Rebiopsie aufgezeigt. Diese Arbeit stellt die Empfehlungen der Sektionen 2 (Endoskopie) und 11 (pneumologische Onkologie) der Deutschen Gesellschaft für Pneumologie (DGP) dar.

Endobronchial ultrasound (EBUS) has revolutionized the diagnosis of lung cancer over the last decade. This minimally invasive diagnostic method has also become increasingly important in the case of other diseases such as sarcoidosis, thereby helping to avoid unnecessary diagnostic interventions. This review article provides an update regarding EBUS and discusses current and future developments of this method.Der endobronchiale Ultraschall (EBUS) hat im letzten Jahrzehnt die Lungenkrebsdiagnostik revolutioniert. Auch bei anderen Erkrankungen wie der Sarkoidose hat der Stellenwert dieses minimalinvasiven diagnostischen Verfahrens erheblich zugenommen und somit geholfen, unnötige diagnostische Operationen zu vermeiden. Dieser Übersichtsartikel stellt ein Update zum EBUS dar und erläutert die aktuellen und zukünftigen Entwicklungen dieses Verfahrens.

Transesophageal ultrasound with the EBUS-bronchoscope (EUS-B) facilitates together with EBUS the staging of lung cancer. Whether EUS-B is feasible for the left adrenal gland (LAG) has not been prospectively evaluated. We run a prospective study with all patients that received EBUS from march-august 2017: Group A with a suspected or history of malignancy; group B without suspicion of malignancy and thoracic lymph nodes. In group A EUS-B-fine needle aspiration (FNA) was performed, if (PET)-CT or EUS-B were suspicious. 4 bronchoscopists (>500 EBUS-procedures) participated. LAG was proven malignant in the following conditions (reference standard): histology by FNA or surgery, size alteration after antitumour therapy. 317 patients were included (group A NSCLC n=179, SCLC n=25, extrathoracic tumor n=13; group B n=96). Identification of the LAG was possible in 274 (87.5%). FNA was performed in 78 patients. Diagnostic yield of 88.5%: metastasis in 9 (11.6%), regular adrenal tissue in 59 (75.6%), myelipoma in 1 (1.3%), non-representative in 9 (11.5%) patients. The reference standard proved 12 malignant LAG. This results in PPV, NPV, sensitivity and specificity of 100%, 99%, 75% and 100% respectively. The blinded radiological assessment resulted in lower PPV, NPV, sensitivity and specificity. After a learning curve of 60 procedures each interventionalist was capable to detect the LAG with a 93% probability. No FNA-related adverse events happened. EUS-B is feasible to locate and characterize the LAG. Even with a longer EBUS experience a learning curve needs to be considered. EUS-B-FNA is safe and can be performed with high sensitivity and specificity. When compared to CT and PET-CT, EUS-B has a higher sensitivity and specificity.

CA extern Epidermal growth factor receptor-tyrosine-kinase inhibitors (EGFR-TKIs) brought a significant revolution in the treatment of non-small-cell lung cancer (NSCLC). In a short period of time, EGFR-TKIs became the standard of treatment for mutation-positive, advanced stage non-squamous NSCLC. In recent years, second- and third-generation EGFR-TKIs are emerging, further widening the clinical use. However, the question of EGFR-TKIs efficiency in the treatment of early stage NSCLC still remains open. Early clinical trials failed to approve the use of EGFR-TKIs in adjuvant setting. The majority of these early trials were performed in unselected NSCLC populations and without standardized biomarker identification. One should certainly not rely solely on these results and dismiss the use of EGFR-TKIs as adjuvant therapy. Many important questions are still unanswered. Most important issues such as stage heterogeneity (IA-IIIA), timing (after or concomitantly with chemotherapy), and type of administration (monotherapy or combination) need to be answered in near future. Adjuvant TKIs in the treatment of lung cancer might offer significant number of advancements. Having in mind the significant duration of response observed in advance disease setting, there could be place for prolongation of response in adjuvant setting potentially, leading to improvement in survival. TKIs could offer less-toxic adjuvant treatment with better efficiency than chemotherapy. However, there is a chronic lack of randomized controlled trials in this field, leading to inability to draw any scientifically sound conclusion with regard to the adjuvant treatment. For now, the use of EGFR-TKIs outside clinical trial setting is not recommended. The purpose of this review is to evaluate current and available data.

Wolfgang Hohenforst-Schmidt,1 Paul Zarogoulidis,2 Joshua Stopek,3 Thomas Vogl,4 Frank Hübner,1 J Francis Turner,5,6 Robert Browning,7 Konstantinos Zarogoulidis,2 Antonis Drevelegas,8 Konstantinos Drevelegas,8 Kaid Darwiche,9 Lutz Freitag,9 Harald Rittger101II Medical Clinic, Coburg Hospital, University of Wuerzburg, Coburg, Germany; 2Pulmonary Department-Oncology Unit, G Papanikolaou General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece; 3Covidien, Jersey City, NJ, USA; 4Department of Diagnostic and Interventional Radiology, Goethe University of Frankfurt, Frankfurt, Germany; 5Division of Interventional Pulmonology, 6Medical Oncology, Cancer Treatment Centers of America, Western Regional Medical Center, Goodyear, AZ, 7Pulmonary and Critical Care Medicine, Interventional Pulmonology, National Naval Medical Center, Walter Reed Army Medical Center, Bethesda, MD, USA; 8Radiology Department, Interbalkan European Medical Center, Thessaloniki, Greece; 9Department of interventional Pneumology, Ruhrlandklinik, University Hospital Essen, University of Essen-Duisburg, Essen, Germany; 10Medical ClinicI, ‘Fuerth Hospital, University of Erlangen, Erlangen, GermanyAbstract: Lung cancer remains the leading cause of death in cancer patients. Severe treatment side effects and late stage of disease at diagnosis continue to be an issue. We investigated whether local treatment using 2-diethylaminoethyl-dextran methyl methacrylate copolymer with p53 (DDMC-p53) with or without cisplatin and/or microwave ablation enhances disease control in BALBC mice. We used a Lewis lung carcinoma cell line to inoculate 140 BALBC mice, which were divided into the following seven groups; control, cisplatin, microwave ablation, DDMC-p53, DDMC-p53 plus cisplatin, DDMC-p53 plus microwave, and DDMC-p53 plus cisplatin plus microwave. Microwave ablation energy was administered at 20 W for 10 minutes. Cisplatin was administered as 1 mL/mg and the DDMC-p53 complex delivered was 0.5 mL. Increased toxicity was observed in the group receiving DDMC-p53 plus cisplatin plus microwave followed by the group receiving DDMC-p53 plus cisplatin. Infection after repeated treatment administration was a major issue. We conclude that a combination of gene therapy using DDMC-p53 with or without cisplatin and microwave is an alternative method for local disease control. However, more experiments are required in a larger model to identify the appropriate dosage profile.Keywords: DDMC, p53, carboplatin, microwave, non-small cell lung cancer

Purpose: Somatostatin is a peptide with a potent and broad antisecretory action, which makes it an invaluable drug target for the pharmacological management of pituitary adenomas and neuroendocrine tumors. Furthermore, somatostatin (SST) receptors (SSTR1, 2A and B, 3, 4 and 5) belong to the G protein coupled receptor family and are overexpressed in tumors. Since, human small-cell lung cancer overexpresses somatostatin receptors (STTR), they could be legitimate targets for treating SCLC.The aim of this study was the evaluation of cytotoxicity of somatostatin in combination with several anticancer drugs in HTB-175 cell line (Small Cell Lung Cancer Cell line that expresses neuron specific enolase). Methods: Docetaxel, Paclitaxel, Carboplatin, Cisplatin, Etoposide, Gemzar, Navelbine, Fluorouracil, Farmorubicin are the chemotherapeutic drugs that we used for the combination before and after adding somatostatin in SCLC cell culture. HTB-175 cell line was purchased from ATCC LGC Standards.At indicated time-point, 48h after the combination, cell viability and apoptosis were measured with Annexin V staining by flow cytometry. Results: Flow cytometry showed that Docetaxel, Paclitaxel, Gemzar and Cisplatin induced apoptosis more when they were added before somatostatin, whereas etoposide induced apoptosis more after somatostatin treatment. Navelbine alone or in combination with somatostatin showed no differences in apoptosis. Farmorubicin showed equal toxicity in all combinations. Fluorouracil and Carboplatin induced apoptosis more when added alone in HTB-175 cell line. However, increased apoptosis was also observed when Carboplatin was administered before somatostatin in higher concentrations. Conclusion: Our results indicated that depending on the drug, somatostatin treatment before or after chemotherapeutic drugs increased apoptosis in small cell lung cancer cells. We suggest that long acting somatostatin analogues could be used as additive and maintenance therapy in combination to antineoplastic agents in SCLC patients.

Historical, the non-small cell lung cancer (NSCLC) was as a united disease entity and the chemotherapy to the metastatic cancer had limited results. Recent studies for the metastatic non-small cell lung cancer led to the ascertainment that the NSCLC does not constitute exclusively a disease entity, but different neoplasms guided from different molecular paths, different biological behavior and at extension requires different confrontation. Thus the new direction for the therapeutic approach of NSCLC is henceforth the most individualized approach based on the activated molecular paths of tumor. Distinct subtypes of NSCLC are driven by a specific genetic alteration, like EGFR, ALK, ROS1 or BRAF mutations, and these genetic alterations are sensitized to the inhibition of specific oncogenic pathways. The benefit from the use of tyrosine kinase inhibitors in patients with EGFR mutations it was confirmed by six randomized studies of phase III that investigated the role of gefitinib, erlotinib and afatinib. In these studies the response rates vary in the impressive percentages from 55% to 86% and were connected with a remarkable median progression free survival of approximately 8 to 13 months, and with better quality of life compared to that of chemotherapy. In early stages NSCLC is needed the individualization of systemic treatment in order to reduce toxicity that is observed in the classic chemotherapy and to impact outcome. The role of EGFR TKI's has been evaluated in the adjuvant chemotherapy in early stage resected NSCLC. The data from these studies suggest that adjuvant TKI therapy might not increase the overall survival, but delay the recurrences. Prospective trials restricted to EGFR or ALK driven NSCLC subsets potentially offering the opportunity for a definitive answer in early disease adjuvant setting (ALCHEMIST) or as induction treatment before stage III chemo-radiotherapy (RTOG 1210/Alliance 31101), are ongoing. Ongoing prospective trials may offer the opportunity for a definitive answer of the role of tyrosine kinase inhibitors in induction treatment before chemo-radiotherapy or in early disease adjuvant therapy.

Endobronchial Ultrasound (EBUS) with the two modalities curved and radial EBUS significantly improved the diagnostics in several pulmonary diseases. The examination and staging of mediastinal and hilar lymph nodes in patients with known or suspected lung malignancy as well as the evaluation of unknown pulmonary or mediastinal lesions can be achieved with minimal invasive means when using EBUS. More invasive surgical procedures for diagnostic purposes can be omitted. The diagnostic yield also increases when EBUS is applied in sarcoidosis or mediastinal lymph node tuberculosis but only to some extend in case of lymphoma. Samples obtained by EBUS-TBNA should be handled efficiently to allow molecular analysis in lung cancer. EBUS is a safe procedure, and complication rate is extremely low. Further advances of the EBUS technology focus on improving analysis of the information provided by the ultrasound image and a better tissue sampling by developing of new EBUS bronchoscopes and TBNA-needles.

Lung cancer is still the leading cause of death among all cancers. During the last 15 years, pharmacogenomics of lung cancer have established targeted therapy with tyrosine kinase inhibitors (TKIs) for epidermal growth factor receptor (EGFR) positive patients in adenocarcinoma or mixed adenosquamus lung cancer patients. However; while novel drugs are released in the market, at the same time novel mutations are observed after tyrosine kinase inhibitor administration. Recently the novel mutation T790 was observed and is highly prevalent in patients already treated with a TKI. A new drug targeting this mutation is already on the market, however; the most important factor for successful treatment in these patients, is adequate tissue re-sampling so that novel mutations can be detected. OA gold - CA extern

BACKGROUND Endobronchial ultrasound with transbronchial needle aspiration (EBUS-TBNA) is a well-established method to assess mediastinal lymph nodes for lung cancer. However, a proportion of patients require further investigation, due to the low negative predictive value (NPV). The objective of this study was to determine whether the assessment of short stature homeobox 2 (SHOX2) DNA methylation level in lymph node tissue obtained by EBUS-TBNA improves the accuracy of mediastinal staging. PATIENTS AND METHODS EBUS-TBNA was carried out for suspicious lymph nodes of 154 patients. Negative or ambiguous histological results were confirmed by surgical means and clinical follow-up over 6 months. EBUS-TBNA was assessed on 80 positive and 85 negative classified lymph nodes and compared with the result of the SHOX2 DNA methylation real-time PCR analysis. Relative methylation measured by delta-delta cycle threshold (ΔΔCt) was used to classify the samples. Clinical performance of the EBUS-TBNA procedure with and without the additional SHOX2 assessment was calculated against the final classification according to the gold standard. RESULTS Based on data from 105 patients, an average 80-fold increase in the SHOX2 methylation level was measured for positive compared with negative lymph nodes. SHOX2 results with a ΔΔCt value of

Lung cancer therapies during the last decade have focused on targeting the genome of cancer cells, and novel routes for administering lung cancer therapies have been investigated for decades. Aerosol therapies for several systematic diseases and systemic infections were introduced into the market a decade ago. One of the main issues of aerosol therapies has been the ability to investigate the deposition of a drug compound throughout the systematic circulation and lymph node circulation. Until now, none of the published studies have efficiently shown the deposition of a chemotherapy pharmaceutical within the lymph node tissue. In our current work we present, for the first time, with the novel CytoViva® (AL, USA) technique, the deposition of cisplatin aerosol therapy in surgically resected stage II lymph nodes from lung cancer patients. Finally, we present the distribution of cisplatin in correlation with the cisplatin concentration in different lymph stations and comment on the possible mechanisms of distribution.

Hemoptysis (coughing up blood) is an extremely alarming situation for the patient, caused by various different underlying diseases. First of all, a peripheral venous catheter should be placed and oxygen should be supplied as patients are threatened by impaired gas exchange caused by the bleeding. Bronchoscopy should be performed immediately, although computed tomography of the thorax may give valuable diagnostic information and should be performed if permitted by the clinical situation. Rigid bronchoscopy should be performed as it allows a broader spectrum of therapeutic options.

Pulmonary metastasectomy has gained the status of a standard treatment for oligometastases of various primaries. Given that the consequences for quality of life (QoL) remain unclear, we initiated this study to characterize the therapy-induced effects of pulmonary metastasectomy on QoL.From 2008 to 2010, patients scheduled for metastasectomy were prospectively evaluated using the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-C30 (EORTC QLQ-C30) and the lung cancer module (LC13) questionnaire and again 3 months later. We analyzed QoL changes over time and looked for sex-specific and age-specific (70 versus70 years) differences.A total of 126 cases were analyzed. The median age of the 73 male and 53 female patients was 59.2 years (range, 24.2 to 83.9). There was no significant change between preoperative and postoperative QoL values for emotional, cognitive, and social functioning. Significant deterioration of QoL items was found for physical functioning (-11.0; p0.001), role functioning (-16.4; p0.001), fatigue (11.1; p0.001), pain (15.0; p0.001), and dyspnea (16.9; p0.001). There were no differences between sexes concerning preoperative and postoperative scores. Younger patients (70 years) had more preoperative symptoms (1.9; p = 0.03) and a worse function (2.2; p = 0.04). A tendency was found for decreased global QoL (-6.0; p = 0.08) in the older age group (70 years) after metastasectomy.Pulmonary metastasectomy can be offered every patient with a chance of cure or prolongation of life because the anticipated midterm changes in QoL are of moderate clinical importance, and the change in global health-related QoL is trivial.

The exhaled breath of lung cancer patients contains volatile organic compounds (VOCs) that differ from those in healthy individuals. These VOCs can be detected with methods such as ion mobility spectrometry (IMS); their origin remains unknown. In 19 patients with lung cancer, exhaled breath was aspirated via the working channel of a flexible bronchoscope from both the tumor-bearing and the opposite lung and analyzed with IMS. IMS measurement through the working channel of a bronchoscope was feasible and safe. In comparison to the opposite lung, we found two peaks that were significantly higher and three peaks that were significantly lower on the IMS of the tumor-bearing site. VOCs differ in concentration depending on the histologic subtype. Our results indicate that VOCs in lung cancer patients are produced locally in or around the tumor, and it is most likely that these VOCs represent underlying metabolic processes of the tumor.

We investigated with seven groups of BALBC mice whether local treatment with DDMC-p53 with or without cisplatin and or microwave ablation enhances disease control. We used Lewis lung carcinoma cell line to inoculate 140 BALBC mice. The mice were divided into the followoing seven groups: a), control, b) cipslatin, c) microwave, d) DDMC-p53, e) DDMC-53 plus cisplatin, f) DDMC-p53 plus microwave and g) DDMC-p53 plus cisplatin plus microwave. Microwave ablation was administered at 20 watts for 10 minutes. Cisplatin administration was 1ml/mg and DDMC-p53 complex delivered was 0.5ml. Increased toxicity was observed in the group of DDMC-p53 plus cisplatin plus microwave following by the group of cisplatin plus microwave. Infection after repeated treatment administration was a major issue. We conclude that combination of gene therapy with DDMC-p53 with or without cisplatin and or microwave is an alternative method for local disease control, however; more experiments are required in a larger model to investigate the proper dosage profile.

Lung cancer accounts for one in five cancer deaths. Broad screening strategies for high-risk populations are unavailable, and the validation of biomarkers for early cancer detection remains a prime interest. Therefore, we investigated the value of circulating U2 small nuclear RNA fragments (RNU2-1f) as a biomarker for diagnosis, prognosis estimation and treatment monitoring in a large lung cancer cohort. We determined RNU2-1f abundance in sera of patients with treatment-naive lung cancer (n = 211, 25.6 % early stage), chronic lung disease (n = 56) and healthy controls (n = 58) by reverse transcription quantitative PCR. Initial levels and changes after one chemotherapy cycle were correlated with treatment outcomes in patient subsets. Relative serum RNU2-1f expression levels (REL) were elevated in lung cancer patients compared with patients with chronic lung disease and healthy controls (p

Background: Mediastinal lung cancer staging is preferably performed via endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA). The assessment of an increasing number of predictive molecular markers for targeted therapy leads to time-delay. Dividing EBUS-TBNA samples allows earlier initiation of targeted therapy. However, it is unknown if this procedure diminishes accuracy and sensitivity of histopathologic and molecular analyses. Objectives: We evaluated accuracy and sensitivity of histopathologic and molecular analyses of divided EBUS-TBNA samples. Methods: EBUS-TBNA was performed in 88 patients with 249 enlarged lymph nodes. Negative or ambiguous histopathological results were confirmed by surgical means and clinical follow-up over 6 months. Samples were transferred onto glass slides, and then manually divided into two equivalent parts. One part was immediately put on dry ice and stored at -80°C before further molecular analyses, the other part was put into formalin before histopathological workup. The accuracy of the histopathological diagnosis and the suitability for molecular analyses were assessed. The amount of needle passes was recorded. Results: Division of EBUS-TBNA samples for simultaneous histopathological and molecular analyses was feasible in all cases . Lymph nodes were punctured an average of 3.18 times. Sensitivity and accuracy of the lymph node staging by EBUS-TBNA were 96.6% and 97.6%, respectively. A molecular test based on cytokeratin-19-mRNA concentration was feasible in 74.1%. Conclusion: Dividing EBUS-TBNA samples instead of taking additional samples provides high accuracy and sensitivity.

Rationale: Cone Beam Computed Tomography imaging has become increasingly important in many fields of interventional therapies. Objective: Lung navigation study which is an uncommon soft tissue approach. Methods: As no effective organ radiation dose levels were available for this kind of Cone Beam Computed Tomography application we simulated in our DynaCT (Siemens AG, Forchheim, Germany) suite 2 measurements including 3D acquisition and again for 3D acquisition and 4 endobronchial navigation maneuvers under fluoroscopy towards a nodule after the 8 th segmentation in the right upper lobe over a total period of 20 minutes (min). These figures reflect the average complexity and time in our experience. We hereby describe the first time the exact protocol of lung navigation by a Cone Beam Computed Tomography approach. Measurement: The hereby first time measured body radiation doses in that approach showed very promising numbers between 0,98-1,15mSv giving specific lung radiation doses of 0,42-0,38 mSv. Main results: These figures are comparable or even better to other lung navigation systems. Cone Beam Computed Tomography offers some unique features for lung interventionists as a realtime 1-step navigation system in an open structure feasible for endobronchial and transcutaneous approach. Conclusions: Due to this low level of radiation exposure Cone Beam Computed Tomography is expected to attract interventionists interested in using and guiding endobronchial or transcutaneous ablative procedures to peripheral endobronchial and other lung lesions.

Kaid Darwiche,1 Paul Zarogoulidis,1,2 Leslie Krauss,3 Filiz Oezkan,1 Robert Fred Henry Walter,1,4 Robert Werner,4 Dirk Theegarten,4 Leonidas Sakkas,5 Antonios Sakkas,5 Wolfgang Hohenforst-Scmidt,6 Konstantinos Zarogoulidis,1 Lutz Freitag11Department of Interventional Pneumology, Ruhrlandklinik, West German Lung Center, University Hospital, University Duisburg-Essen, Essen, Germany; 2Pulmonary Department, Oncology Unit, G Papanikolaou General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece; 3CytoViva, Inc, Auburn, AL, USA; 4Institute of Pathology and Neuropathology, University Hospital of Essen, University of Duisburg-Essen, Essen, Germany; 5Pathology Department, G Papanikolaou General Hospital, Thessaloniki, Greece; 6II Medical Department, Coburg Regional Clinic, University of Wuerzburg, Coburg, GermanyBackground: There are currently many techniques and devices available for the diagnosis of lung cancer. However, rapid on-site diagnosis is essential for early-stage lung cancer, and in the current work we investigated a new diagnostic illumination nanotechnology.Methods: Tissue samples were obtained from lymph nodes, cancerous tissue, and abnormal intrapulmonary lesions at our interventional pulmonary suites. The following diagnostic techniques were used to obtain the samples: endobronchial ultrasound bronchoscopy; flexible bronchoscopy; and rigid bronchoscopy. Flexible and rigid forceps were used because several of the patients were intubated using a rigid bronchoscope. In total, 30 tissue specimens from 30 patients were prepared. CytoViva® illumination nanotechnology was subsequently applied to each of the biopsy tissue slides.Results: A spectral library was created for adenocarcinoma, epidermal growth factor receptor mutation-positive adenocarcinoma, squamous cell carcinoma, usual interstitial pneumonitis, nonspecific interstitial pneumonitis, typical carcinoid tumor, sarcoidosis, idiopathic pulmonary fibrosis, small cell neuroendocrine carcinoma, thymoma, epithelioid and sarcomatoid mesothelioma, cryptogenic organizing pneumonia, malt cell lymphoma, and Wegener's granulomatosis.Conclusion: The CytoViva software, once it had created a specific spectral library for each entity, was able to identify the same disease again in subsequent paired sets of slides of the same disease. Further evaluation of this technique could make this illumination nanotechnology an efficient rapid on-site diagnostic tool.Keywords: lung cancer, endobronchial ultrasound, spectral imaging, diagnosis

Konstantinos Zarogoulidis,1 Efimia Boutsikou,1 Paul Zarogoulidis,1,2 Kaid Darwiche,2 Lutz Freitag,2 Konstantinos Porpodis,1 Dimitrios Latsios,1 Theodoros Kontakiotis,1 Haidong Huang,3,4 Qiang Li,4 Wolfgang Hohenforst-Schmidt,5 Maria Kipourou,1 J Francis Turner,6 Dionysios Spyratos11Pulmonary Department, “G Papanikolaou” General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece; 2Department of Interventional Pneumology, Ruhrlandklinik, West German Lung Center, University Hospital, University Duisburg-Essen, Essen, Germany; 3Bronchoscopy and Interventional Pulmonology, Pulmonary and Critical Care Medicine, Henry Ford Hospital, Wayne State University, School of Medicine, Detroit, MI, USA; 4Department of Respiratory Diseases, Changhai Hospital/First Affiliated Hospital of the Second Military Medical University, Shanghai, People’s Republic of China; 5II Medical Clinic, “Coburg” Regional Hospital, University of Wuerzburg, Coburg, Germany; 6Pulmonary Medicine, University of Nevada School of Medicine, National Supercomputing Center for Energy and the Environment University of Nevada, Las Vegas, NV, USABackground: To evaluate the benefit of second-line chemotherapy with platinum-based treatment in patients with recurrent small cell lung cancer (SCLC).Patients and methods: A total of 535 patients continued with follow-up or best supportive care if needed, and 229 patients who progressed after the completion of first-line chemotherapy were treated with second-line chemotherapy at the time of progression. In total, 103/229 patients received paclitaxel 190 mg/m2 and carboplatin 5.5 area under the curve while 126/229 patients received etoposide 200 mg/m2 and carboplatin 5.5 area under the curve every 28 days.Results: Patients administered second-line chemotherapy lived significantly longer, with a median survival of 422 days compared to 228 days in patients with best supportive care alone (P

Lung cancer first line treatment has been directed from the non-specific cytotoxic doublet chemotherapy to the molecular targeted. The major limitation of the targeted therapies still remains the small number of patients positive to gene mutations. Furthermore, the differentiation between second line and maintenance therapy has not been fully clarified and differs in the clinical practice between cancer centers. The authors present a segregation between maintenance treatment and second line and present a possible definition for the term “maintenance” treatment. In addition, cancer cell evolution induces mutations and therefore either targeted therapies or non-specific chemotherapy drugs in many patients become ineffective. In the present work pathways such as epidermal growth factor, anaplastic lymphoma kinase, met proto-oncogene and PI3K are extensively presented and correlated with current chemotherapy treatment. Future, perspectives for targeted treatment are presented based on the current publications and ongoing clinical trials.

Lung cancer still remains one of the leading causes of death among cancer patients. Although novel targeted therapies have been established in everyday treatment practice, and conventional platinum-based doublets have demonstrated effective results regarding overall and progression-free survival, we have still failed to achieve long-term survival. Therefore, several strategies of applying locoregional therapy are under investigation. Aerosol chemotherapy is already under investigation and, taking this a step further, aerosol gene therapies with multiple delivery systems are being developed. Several efforts have demonstrated its efficiency and effectiveness, but there are still multiple factors that have to be considered and combined to achieve an overall more effective multifunctional treatment. In the current review, we present data regarding aerosol delivery systems, transporters, carriers, vectors, genes, toxicity, efficiency, specificity, lung microenvironment and delivery gene therapy systems. Finally, we present current studies and future perspectives.

Konstantinos Zarogoulidis,1 Eftimios Ziogas,1 Efimia Boutsikou,1 Paul Zarogoulidis,1,2 Kaid Darwiche,2 Theodoros Kontakiotis,1 Kosmas Tsakiridis,3 Konstantinos Porpodis,1 Dimitrios Latsios,1 Olga Chatzizisi,4 Ilias Karapantzos,5 Qiang Li,6 Georgios Kyriazis21Pulmonary Department, "G Papanikolaou" General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece; 2Department of Interventional Pneumology, Ruhrlandklinik, West German Lung Center, University Hospital, University Duisburg-Essen, Essen, Germany; 3Thoracic Surgery Department, "Saint Luke" Private Hospital, Panorama, Thessaloniki, Greece; 4Immunology Department, "G Papanikolaou" General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece; 5Ear, Nose and Throat Department, "Saint Luke" Private Hospital, Panorama, Thessaloniki, Greece; 6Respiratory Diseases Department, Shanghai hospital, Second Military Medical University, Shanghai, People's Republic of ChinaPurpose: To evaluate the effect of immunotherapy on response, survival, and certain immune markers in patients with small cell lung cancer (SCLC) who are receiving chemotherapy.Patients and methods: Patients with SCLC (n = 164) were assigned to receive either chemotherapy alone (group A) or a combination of chemotherapy and immunotherapy as follows: interferon α (IFN-α; 3 million IU) 3 times per week (group B); IFN-γ (3 million IU) 3 times per week (group C); and IFN-α and IFN-γ (1.5 million IU of each) 3 times per week (group D). Chemotherapy was the same for all groups and consisted of eight cycles with carboplatin 5.5 mg/m2 intravenously on day 1, ifosfamide 3.5 mg/m2 intravenously on day 1, and etoposide 200 mg/m2 total dose taken orally on days 1 through 3, every 28 days. Patients completing chemotherapy were restaged, and those who were found to have limited disease received primary site and prophylactic cranial irradiation. Immunotherapy was continued throughout these treatments and during the follow-up period. Blood was taken before each course of chemotherapy and during follow-up to measure CD3+ lymphocytes, CD3+CD4+ lymphocytes, CD3+CD8+ lymphocytes, natural killer cells, and natural killer T cells.Results: Differences in response and survival were not significantly different when all patients were considered. However, among patients with limited disease, Kaplan–Meier analysis disclosed a survival benefit for group B (P < 0.05). The analysis of immunologic measurements revealed that the improvement of immune markers was always accompanied by clinical improvement, whereas deterioration of all markers was accompanied by disease progression (result not statistically significant except for group C; P < 0.05).Conclusion: Among cytokines used in the study, only IFN-α seems to confer a survival benefit to patients with SCLC with limited disease. However, immunotherapy remains a challenge in the treatment of lung neoplasms and should be further explored.Keywords: interferon, SCLC, lung cancer, immunomodifiers

Anastasios Kallianos,1 Sotirios Tsimpoukis,2 Paul Zarogoulidis,3 Kaid Darwiche,4 Andriani Charpidou,2 Ilias Tsioulis,3 Georgia Trakada,5 Konstantinos Porpodis,3 Dionysios Spyratos,3 Athanasios Panoutspoulos,5 Lemonia Veletza,5 Konstantinos Kostopoulos,5 Charalampos Kostopoulos,4 Ilias Karapantzos,6 Kosmas Tsakiridis,7 Wolfgang Hohenforst-Schmidt,8 Konstantinos Zarogoulidis,3 Aggeliki Rapti,1 Konstantinos Syrigos21Second Pulmonary Clinic, Sotiria Hospital, 2Oncology Unit, Sotiria Hospital, Athens, Greece; 3Oncology Unit, G Papanikolaou General Hospital, Thessaloniki, Greece; 4Interventional Unit, Ruhrland Clinic, Essen, Germany; 5Pulmonary Laboratory, Alexandra Hospital, Athens, Greece; 6Ear, Nose, and Throat Department, Saint Luke Private Hospital, 7Cardiothoracic Department, Saint Luke Private Hospital, Thessaloniki, Greece; 8II Medical Clinic, Hospital of Coburg, Coburg, GermanyAbstract: Nitric oxide (NO) is a marker of airway inflammation and indirectly a general indicator of inflammation and oxidative stress. NO is a contributing factor in lung cancer at an early stage and also after chemotherapy treatment of lung cancer. We studied whether exhaled NO levels were altered by three cycles of chemotherapy at diagnosis and after chemotherapy, and whether, directly or indirectly, these changes were related to the course of disease. Also, a correlation of NO levels with other markers of inflammation was performed. We studied 42 patients diagnosed early: 26 men and 16 women with lung cancer. We analyzed blood tests for control of inflammatory markers, functional pulmonary tests, and alveolar exhaled NO. We recorded a decrease in exhaled NO after three cycles of chemotherapy in all patients, regardless of histological type and stage: there were 42 patients with mean 9.8 NO after three cycles (average 7.7). Also, a strong correlation appeared between NO measurements before and after chemotherapy and C-reactive protein (P < 0.05, r = 0.42, before) and (P < 0.045, r = 0.64, after). NO alveolar measurement as an indicator of airway inflammation indicates response to chemotherapy in lung cancer. Also, the inflammatory process in lung cancer was confirmed and indicated response to chemotherapy through an index that is sensitive to inflammatory disease of the airways.Keywords: nitric oxide, lung cancer, inflammation

Purpose Multiple investigational drugs are currently explored in cancer patient populations defined by specific biomarkers. This demands a new process of patient selection for clinical trials. Patients and methods Starting January 1, 2012, preemptive biomarker profiling was offered at the West German Cancer Center to all patients with advanced non-small-cell lung (NSCLC) or colorectal cancer (CRC), who met generic study inclusion criteria. Tumour specimens were subjected to prespecified profiling algorithms to detect ‘actionable biomarkers' by amplicon sequencing, in situ hybridisation and immunohistochemistry. The clinical course was closely monitored to offer trial participation whenever applicable. Results Within 12 months, 267 patients (188 NSCLC, 79 CRC) were profiled. Estimated additional cost for biomarker profiling was 219615.51 EUR excluding histopathology workup and administration. The most prevalent biomarkers in pulmonary adenocarcinoma were KRAS mutations (29%), loss of PTEN expression (18%), EGFR mutations (9%), HER2 amplification (5%) and BRAF mutations (3%), while the prevalence of ALK translocations and PIK3CA mutations was extremely low. In pulmonary squamous cell carcinoma FGFR1 amplifications were found in 15%, PTEN expression was lost in 20% and DDR2 was mutated in a single case. KRAS mutations (41%) predominated in CRC, followed by loss of PTEN expression (16%), PIK3CA (5%) and BRAF (5%) mutations. So far 13 patients (5%) have entered biomarker-stratified clinical trials. Therapeutic decisions for approved drugs were guided in another 45 patients (17%). Conclusion Preemptive biomarker profiling can be implemented into the diagnostic algorithm of a large Comprehensive Cancer Center. Substantial investments in diagnostics and administration are required.

Konstantinos Zarogoulidis,1 Dimitrios Latsios,1 Konstantinos Porpodis,1 Paul Zarogoulidis,1,2 Kaid Darwiche,2 Nick Antoniou,1 Wolfgang Hohenforst-Schmidt,3 Ellada Eleftheriadou,1 Efimia Boutsikou,1 Theodoros Kontakiotis11Pulmonary Department-Oncology Unit, “G Papanikolaou” General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece; 2University Pulmonary Department- Interventional Unit, Ruhrlandklinik, University, University of Duisburg-Essen, Essen, Germany; 3II Medical Clinic, Hospital of Coburg, University of Wuerzburg, Coburg, GermanyBackground: Many patients with limited disease (LD) behave similarly to those with extensive disease (ED) from a prognostic point of view. On the other hand, a proportion of patients with ED small-cell lung cancer (SCLC) behave similarly to those with LD.Patients and methods: In this retrospective study analysis, 764 patients with proven SCLC were included and managed with the same therapeutic protocols. Of these patients, 278 (36.4%) had LD, while 486 (63.6%) had ED.Results: No statistically significant difference was observed for survival for IA and IB disease stages (P = 0.254) and between IIA and IIB stages (P = 0.256) according to the new tumor, node, metastasis (TNM) staging classification classification. In addition, no statistical significant difference was observed for survival between patients with (IIA + IIB) and IIIA (P = 0.951), (IIA + IIIA, P = 0.658), and (IIB + IIIA, P = 0.573) stages. Statistical significant difference was observed for survival among the LD SCLC patients with (IA + IB), (IIA + IIB + IIIA), and IIIB stages (P < 0.001). Similarly, statistical significance was observed for ED SCLC patients with (IIA + IIB + IIIA), IIIB, and IV stages (P < 0.001).Conclusions: Although stratification of SCLC patients in LD and ED is generally satisfactory, the TNM staging system is recommended for more detailed prognostic information and treatment evaluation in these patients.Keywords: small-cell lung cancer, staging, lung cancer

Eftimia Boutsikou,1 Theodoros Kontakiotis,1 Paul Zarogoulidis,1 Kaid Darwiche,2 Ellada Eleptheriadou,1 Konstantinos Porpodis,1 Grammati Galaktidou,3 Leonidas Sakkas,4 Wolfgang Hohenforst-Schmidt,5 Kosmas Tsakiridis,6 Theodoros Karaiskos,7 Konstantinos Zarogoulidis11Pulmonary Department-Oncology Unit, G Papanikolaou General Hospital, Aristotle University of Thessaloniki, Greece; 2University Pulmonary Department-Interventional Unit, Ruhrland Klinic, University of Duisburg-Essen, Essen, Germany; 3Theagenio Anticancer Institute Research Laboratory, 4Department of Pathology, G Papanikolaou Hospital, Thessaloniki, Greece; 5II Medical Clinic, Hospital Coburg, University of Wuerzburg, Coburg, Germany; 6Cardiothoracic Surgery Department, Saint Luke Private Hospital, Panorama, 7Cardiothoracic Surgery Department, G Papanikolaou General Hospital, Aristotle University of Thessaloniki, Thessaloniki, GreeceBackground: Bevacizumab and erlotinib have been demonstrated to prolong overall survival in patients with non-squamous non-small cell lung cancer (NSCLC). We designed a four-arm Phase III trial to evaluate the efficacy and toxicity of the combination of docetaxel, carboplatin, bevacizumab, and erlotinib in the first-line treatment of patients with NSCLC.Methods: A total of 229 patients with stage IIIb/IV non-squamous NSCLC were treated with two cycles of carboplatin (area under the concentration-time curve 5.5) and docetaxel 100 mg/m2 as chemotherapy. After completion of two treatment cycles, patients were evaluated for response and divided into four groups: 61/229 continued with four more cycles of chemotherapy (control group), 52/229 received chemotherapy plus erlotinib 150 mg daily, 56/229 received chemotherapy plus bevacizumab 7.5 mg/kg, and 60/229 were treated with the combination of chemotherapy, erlotinib, and bevacizumab until disease progression. The primary endpoint was overall survival.Results: Over 4 years of follow-up, there was no statistically significant difference in survival and time to progression between the four treatment groups. After two cycles of chemotherapy, responders and nonresponders were divided according to their response in order to examine the role of initial response as an independent factor in survival and response when a biological agent is combined with chemotherapy. Nonresponders, who received additional therapy with bevacizumab or combination therapy, had a survival benefit [657 days (95% confidence interval 349–970) and 681 days (95% confidence interval 315–912), respectively], which was statistically significant compared with continuation of cytotoxic chemotherapy (P < 0.001). The combination therapy had a safety profile comparable with that of bevacizumab and erlotinib taken individually.Conclusion: Administration of bevacizumab and erlotinib in combination with first-line chemotherapy, followed by bevacizumab and erlotinib monotherapy as maintenance, showed promising results in patients with NSCLC, with reduced toxicity as compared with chemotherapy alone, but did not translate into longer overall survival.Keywords: vascular endothelial growth factor, epidermal growth factor receptor, erlotinib, bevacizumab, non-small cell lung cancer

Chemotherapy drugs have still the major disadvantage of non-specific cytotoxic effects. Although, new drugs targeting the genome of the tumor are already in the market, doublet chemotherapy regimens still remain the cornerstone of lung cancer treatment. Novel modalities of administration are under investigation such as; aerosol, intratumoral and intravascular.In the present study five chemotherapy drugs; paclitaxel, docetaxel, gemcitabine, carboplatin and cisplatin were nebulized with three different jet nebulizers (Maxineb(®), Sunmist(®), Invacare(®)) and six different residual cups at different concentrations. The purpose of the study was to identify the "ideal" combination of nebulizer-residual cup design-drug-drug loading for a future concept of aerosol chemotherapy in lung cancer patients. The Mastersizer(®) 2000 was used to evaluate the aerosol droplet mass median aerodynamic diameter.The drug, nebulizer and residual cup design greatly influences the producing droplet size (p0.005, in each case). However; the design of the residual cup is the most important factor affecting the produced droplet size (F=834.6, p0.001). The drug loading plays a vital role in the production of the desired droplet size (F=10.42, p0.001). The smallest droplet size was produced at 8 ml loading (1.26 μm), while it remained the same at 2, 4 and 6 mls of drug loading.The ideal nebulizer would be Maxineb(®), with a large residual cup (10 ml maximum loading capacity) and 8 mls loading and the drug with efficient pulmonary deposition would be docetaxel.

Inhaled chemotherapy was first used more than 30 years ago. Since then, numerous chemotherapeutic agents have been used in either in vitro or in vivo studies. Several aspects of the methodology of the drug administration have been thoroughly demonstrated and explained. However, the safety concerns of these studies were not thoroughly investigated and different results regarding the same drug formulations have been reported. There are cases where the studies failed to demonstrate the long-term effects of the chemotherapeutic drug formulations to the lung parenchyma. Acute and latent effects observed in a small number of human trial studies are still under investigation of inhaled chemotherapy administration. This review provides data regarding all up-to-date inhaled chemotherapy studies and presents the methodological parameters of the safety measures incorporated. In addition, a commentary regarding the safety concerns for the medical staff participating in these studies will be presented.