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PDF file - 126K, PF-2341066 treatment upfront decreases tumorogenesis and local invasion in H69M xenograft.

PDF file - 110K, Karyotypic and gene expression differences between H69 and H69M.

PDF file - 119K, Supplementary Table 1. Patients� characteristics. Supplementary Table 2. Biomarker expression in human samples. Supplementary Table 3. Spearman correlation between biomarkers� in cases showing expression. Supplementary Table 4. Association of Met and p-Met with other biomarkers. Supplementary Table 5. Cox regression analysis for overall survival for each variable adjusted for performance status (PS) and stage. Supplementary Table 6. Biomarker status in biopsies pre-treatment and at relapsed disease.

PDF file - 441K, Cellular and molecular changes induced by Met inhibitors.

PDF file - 354K, PF-2341066 sensitizes H841-derived xenografts cells to chemotherapy.

PDF file - 40K, Kaplan-Meier curves for overall survival univariate analysis according to biomarker status.

PDF file - 163K, H69M2 are phenotypically mesenchymal and more tumorogenic.

Background Stroma, mainly composed by fibroblasts, extracellular matrix (ECM) and vessels, may play a role in tumorigenesis and cancer progression. Chronic Obstructive Pulmonary Disease (COPD) is an independent risk factor for LC. We hypothesized that markers of fibroblasts, ECM and endothelial cells may differ in tumors of LC patients with/without COPD. Methods Markers of cultured cancer-associated fibroblasts and normal fibroblasts [CAFs and NFs, respectively, vimentin and alpha-smooth muscle actin (SMA) markers, immunofluorescence in cultured lung fibroblasts], ECM, and endothelial cells (type I collagen and CD31 markers, respectively, immunohistochemistry) were identified in lung tumor and non-tumor specimens (thoracotomy for lung tumor resection) from 15 LC-COPD patients and 15 LC-only patients. Results Numbers of CAFs significantly increased, while those of NFs significantly decreased in tumor samples compared to non-tumor specimens of both LC and LC-COPD patients. Endothelial cells (CD31) significantly decreased in tumor samples compared to non-tumor specimens only in LC patients. No significant differences were seen in levels of type I collagen in any samples or study groups. Conclusions Vascular endothelial marker CD31 expression was reduced in tumors of non-COPD patients, while type I collagen levels did not differ between groups. A rise in CAFs levels was detected in lung tumors of patients irrespective of airway obstruction. Low levels of CD31 may have implications in the overall survival of LC patients, especially in those without underlying airway obstruction. Identification of CD31 role as a prognostic and therapeutic biomarker in lung tumors of patients with underlying respiratory diseases warrants attention.

Resumen En el ano 2011 se inicio un proyecto conjunto entre la Sociedad Espanola de Oncologia Medica (SEOM) y la Sociedad Espanola de Anatomia Patologica (SEAP) para establecer unas recomendaciones basadas en la evidencia actual con respecto a la determinacion de biomarcadores en pacientes con carcinoma de pulmon de celula no pequena avanzado. Al ser un area en continua evolucion, estas recomendaciones se han actualizado previamente en 2012 y 2015, y ahora en 2019. Con la evidencia que existe hoy en dia, las determinaciones obligatorias en cualquier paciente con este tipo de carcinoma de pulmon avanzado son las mutaciones de EGFR y BRAF, los reordenamientos de ALK y ROS1, y la expresion de PD-L1. La creciente necesidad que existe para estudiar otros biomarcadores emergentes promueve el uso de forma rutinaria de la secuenciacion masiva (next-generation sequencing, NGS). Continua siendo un reto coordinar a todos los profesionales implicados y priorizar las determinaciones y las tecnologias mas adecuadas en cada caso.

Non-small cell lung cancer (NSCLC) is one of the oncological entities with the greatest evolution in molecular diagnosis due to the large number of diagnostic biomarkers and new treatments approved by international regulatory agencies. An accurate, early diagnosis using the least amount of tissue is the goal for the establishing and developing precision medicine for these patients. Rapid on-site evaluation (ROSE) provides cytological samples of optimal quantity and quality for a complete diagnosis of NSCLC. The usefulness of cytological samples has been demonstrated, not only for massive parallel sequencing but also for the quantification of the expression of programmed death-ligand 1 (PD-L1) and tumour mutational burden (TMB). Pre-analytical, analytical, and post-analytical recommendations are made for the management and appropriate use of cytological samples in order to obtain all the information necessary for the diagnosis and treatment of patients with NSCLC according to current quality parameters.

Biopsy site changes in mediastinal lymph nodes (LNs) attributable to prior endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) have not been studied in a systematic manner. Twenty-four contributors from 14 institutions in 5 countries collaborated via social media (Twitter) to retrospectively review consecutive cases of resected mediastinal LNs from patients with prior EBUS-TBNA. Resected LNs were reexamined by submitting pathologists for changes attributable to EBUS-TBNA. Patients who received neoadjuvant therapy were excluded. Cases with suspected biopsy site changes underwent central review by 5 pathologists. A total of 297 mediastinal LN resection specimens from 297 patients (183 male/114 female, mean age: 65 y, range: 23 to 87) were reviewed. Biopsy site changes were most common in station 7 (10 cases) followed by 11R, 4R, and 10R, and were found in 34/297 (11.4%) cases, including displacement of tiny cartilage fragments into LN parenchyma in 26, intranodal or perinodal scars in 7, and hemosiderin in 1. Cartilage fragments ranged from 0.26 to 1.03 mm in length and 0.18 to 0.62 mm in width. The mean interval between EBUS-TBNA and LN resection was 38 days (range: 10 to 112) in cases with biopsy site changes. A control group of 40 cases without prior EBUS-TBNA, including 193 mediastinal LN stations, showed no evidence of biopsy site changes. Biopsy site changes are identified in a subset of resected mediastinal LNs previously sampled by EBUS-TBNA. The location of the abnormalities, temporal association with prior EBUS-TBNA, and the absence of such findings in cases without prior EBUS-TBNA support the contention that they are caused by EBUS-TBNA.

Simple Summary Many people still die of lung cancer (LC), a disease that is mainly related to cigarette smoking. Smokers may also develop chronic obstructive pulmonary disease (COPD). COPD is a risk factor per se for LC. Cigarette smoking and other chemicals injure DNA on a daily basis. A repair mechanism based on PARP-1 and PARP-2 activity can restore damaged DNA to keep cells alive. However, cancer cells also take advantage of this mechanism to survive. Fifteen LC-COPD and 15 LC patients were enrolled in this study to elucidate whether COPD influences DNA damage-dependent PARP activity in lung tumors. DNA damage, PARP activity, PARP-1 and PARP-2 expression were analyzed in tumor and non-tumor lungs obtained during surgical resection of the lung tumors. DNA damage and PARP activity were increased only in tumors in LC-COPD patients. However, PARP-1 and PARP-2 expression decreased in tumors of both patient groups. LC patients with COPD may benefit from PARP inhibitor therapies. Abstract (1) Background: Lung cancer (LC) is a major leading cause of death worldwide. Poly (ADP-ribose) polymerase (PARP)-1 and PARP-2 are key players in cancer. We aimed to assess PARP-1 and PARP-2 expression and activity and DNA damage in tumors and non-tumor lungs from patients with/without chronic obstructive pulmonary disease (COPD). (2) Methods: Lung tumor and non-tumor specimens were obtained through video-assisted thoracoscopic surgery (VATS) in LC patients with/without underlying COPD (two groups of patients, n = 15/group). PARP-1 and PARP-2 expression (ELISA), PARP activity (PARP colorimetric assay kit) and DNA damage (immunohistochemistry) levels were identified in all samples. (3) Results: Both PARP-1 and PARP-2 expression levels were significantly lower in lung tumors (irrespective of COPD)compared to non-tumor specimens, while DNA damage and PARP activity levels significantly increased in lung tumors compared to non-tumor specimens only in LC-COPD patients. PARP-2 expression was positively correlated with smoking burden in LC-COPD patients. (4) Conclusions: In lung tumors of COPD patients, an overactivation of PARP enzyme was observed. A decline in PARP-1 and PARP-2 protein expression was seen in lung tumors irrespective of COPD. Other phenotypic features (airway obstruction) beyond cancer may account for the increase in PARP activity seen in the tumors of patients with underlying COPD.

Immune microenvironment plays a role in the development of lung cancer (LC). We hypothesized that immune profile of B and T cells may differ in tumors of LC patients with and without COPD and may also influence survival. Objectives: 1) To analyze levels of tertiary lymphoid structures (TLSs), B and T cells in tumor and non-tumor (control samples) lung specimens of LC patients with/without COPD and 2) to analyze the influence of those biological markers in the patients´10-year survival. TLSs (numbers and area), B (CD20), and T (CD3) cells were identified in both tumor and non-tumor specimens (thoracotomy) from 90 LC-COPD patients and 43 LC-only patients (immunohistochemistry). Survival was analyzed in all 133 patients. Immune profile in tumors of LC-COPD versus LC: The number of TLSs significantly decreased in tumors of LC-COPD compared to LC patients. No significant differences were observed in tumors between LC-COPD and LC for B or T cells. In tumors compared to non-tumor specimens, a significant rise in TLSs was observed in LC (numbers and area) and LC-COPD (area), T cell counts declined in tumors of LC, while B cell counts increased in tumors of both LC and LC-COPD patients. Survival: In LC-COPD patients: lower numbers of TLSs and greater numbers of B cells were associated with worse survival. In LC patients: lower levels of T cells were associated with longer survival rates. TLSs, B cells and T cells are differentially expressed in tumors of LC-COPD from that in LC-only patients. Method: FIS 18/00075 & CIBERES (FEDER, ISC-III), SEPAR 2018, unrestricted research grant from Menarini SA 2018.

Introduction: In two previous cohorts we demonstrated that EGFR/KRAS mutations can be present in non-tumoral lung tissue (NTLT) regardless tumor genomic status. In cohort one (J Clin Med. 2019) the presence of these mutations was associated with a worse prognosis (disease-free survival), while in cohort two the presence of NTLT mutations in EGFR and KRAS was demonstrated even in the absence of mutations in the primary tumor (no differences in prognosis were found due to small sample size). The objective of this study is to analyze the 5-year prognosis of both cohorts and assess whether the presence of driver-mutations in NTLT is associated with a higher overall mortality in patients with localized ADC. We present the preliminary results Methods: 78 patients were included (47 from the first cohort and 31 from the second one). After tumor genotyping, the EGFR/KRAS status of the NTLT was performed with real-time PCR Results: two patients did not continued the follow-up. The remaining completed the 36-months follow-up and 77% have completed the 5-year follow-up at the time of the analysis. 13 (16.7%) patients had mutations in the EGFR and/or KRAS in the NTLT (M-NTLT group). At 60 months, 53.8% of M-NTLT group patients had distant recurrence vs 32.3%. Overall mortality was higher in the M-NTLT group: average survival of 40.5 months vs 54.8 (-14.3 months, Log-rank p=0.003). These results are independent of the type of mutation, cancer treatment and preoperative stage Conclusion: based on two single-center cohorts, we can conclude that in patients with localized ADC, having EGFR or KRAS mutations in NTLT is associated with a higher risk of death. These data reaffirm the results of our previous studies

Background Twitter is an expanding social media network among cytopathologists to share knowledge. Tweets are made up of text which may also include images or video. All tweets labeled under a hashtag can be tracked. The #FNAFriday hashtag was created in 2015 by one of the authors (X.J.) to build a community of individuals, to educate and share interesting cases, and highlight a variety of diagnoses with FNA specimens. Methods We retrospectively extracted all tweets labeled with #FNAFriday from April 2015 to mid-February 2019 (47 mo) using the Twitter search engine. The data point included: author, number of figures, type of cytology-stain, use of immunocytochemistry, histochemistry or molecular techniques, and the subspeciality. The educational content was categorized as: live-tweeting, training activities, and publication references. The number if comments, retweets and likes was also recorded. Results A total of 349 original tweets using #FNAFriday were tracked with an average of 7.43 tweets/month. We describe the "top three" countries with most tweets, active users and subspecialties. The most frequent stain was Papanicolau and part of the content of the tweets was using cellblock (14.04%), histologic correlation (10.03%), immunocytochemistry (8.60%), molecular tests (2.01%), gross pictures (4.58%), and radiologic pictures (3.4%). Conclusion The presence of cytopathologists on Twitter who want to share their cases has increased. The weekly FNAFriday label with other cytology hashtags is a specific keyword for those interested in the field.

Background More than 60% of patients with lung cancer are diagnosed at advanced stages. The introduction of targeted therapies requires molecular diagnosis to guide treatment. The aim of this study was to evaluate the feasibility of performing mutational analysis with brushing specimens obtained by radial-miniprobe endobronchial ultrasound (R-EBUS) plus fluoroscopy-guided bronchoscopy in patients with peripheral pulmonary adenocarcinoma. Methods Brushing specimens were deposited on cytological slides and were conserved in Roswell Park Memorial Institute (RPMI) culture medium. DNA was isolated to perform a mutational analysis with real-time quantitative polymerase chain reaction and Sanger sequencing for epidermal growth factor receptor (EGFR) and Kirsten rat sarcoma viral oncogene homolog (KRAS). Results Thirty patients with adenocarcinoma were prospectively included. In 100% of the patients, the molecular study was viable with brushing specimens. In 16 (53.3%), KRAS or EGFR mutations were detected: 10 KRAS mutations (33.3%) and 6 EGFR mutations (20%). In a comparison with histological samples, a correlation of 86.6% was detected, and only 2 patients with wild-type results from brushing specimens presented with an EGFR mutation in histological samples. Conclusions Brushing specimens conserved in RPMI medium and obtained by R-EBUS plus fluoroscopy-guided bronchoscopy are valid for molecular studies. They allow the detection of EGFR/KRAS mutations in patients with peripheral adenocarcinoma. In addition, invasive techniques are avoided, the risk of complications is reduced, and the obtained samples are optimized.

Introduction Bronchoalveolar lavage (BAL) analysis has been proposed as an objective technique for confirming asbestos exposure. However, the reliability and diagnostic yield of this procedure has not been studied in Spain. The aim of this study was to assess the usefulness of the analysis of asbestos bodies (AB) in bronchoalveolar lavage (BAL) for the diagnosis of asbestos-related diseases (ARD). Methods BAL samples from 72 patients (66 male, mean age 66 years) undergoing bronchoscopy were analyzed. Lung tissue from 23 of these patients was also analyzed. Asbestos exposure was assessed by anamnesis and a review of the patient's medical records. BAL and lung samples were processed and AB count was determined by light microscopy. The accepted threshold value to diagnose asbestos-related diseases was 1 AB/ml BAL or 1000 AB/gr dry tissue. Results Thirty-nine patients reported exposure to asbestos. Of these, 13 (33%) presented AB values above 1 AB/ml BAL. In the 33 non-exposed patients, 5 (15%) presented AB values above 1 AB/ml BAL. There was a significant difference between the AB levels of exposed and non-exposed patients (P =.006). The ROC curve showed that a value of 0.5 AB/ml BAL achieved the most satisfactory sensitivity, 46%, and a specificity of 83%. The correlation between AB levels in BAL and lung was 0.633 (P =.002). Conclusions BAL study provides objective evidence of exposure to asbestos. The good correlation between the AB counts in BAL and lung tissue indicates that both techniques are valid for the analysis of asbestos content.

Resumen Introduccion El analisis del lavado broncoalveolar (LBA) se ha propuesto como tecnica objetiva para certificar la exposicion a amianto. Sin embargo, la fiabilidad y rendimiento diagnostico de este procedimiento diagnostico no se han analizado en Espana. El proposito de este estudio fue evaluar la utilidad del analisis de cuerpos de amianto (CA) en el LBA para el diagnostico de enfermedades relacionadas con el amianto (ERA). Metodos Se analizaron muestras de LBA de 72 pacientes (66 varones, edad media de 66 anos) sometidos a broncoscopia. Tambien se analizo el tejido pulmonar de 23 de estos pacientes. La exposicion al amianto se evaluo a partir de la anamnesis y la revision de las historias clinicas de los pacientes. Las muestras de LBA y de tejido pulmonar se procesaron, y la cantidad de CA se determino mediante microscopia optica. El valor umbral aceptado para diagnosticar una enfermedad relacionada con el amianto fue de 1 CA/ml de LBA o 1.000 CA/g de tejido seco. Resultados Treinta y nueve pacientes refirieron exposicion a amianto. En 13 (33%) de estos pacientes, los niveles de CA fueron superiores a 1 CA/ml de LBA. De los 33 pacientes no expuestos, los valores de CA fueron superiores a 1 CA/ml de LBA en 5 casos (15%). La diferencia entre los niveles de CA de los pacientes expuestos y los no expuestos fue significativa (p = 0,006). La curva ROC indico que el nivel de 0,5 CA/ml de LBA era el que alcanzaba mayor sensibilidad (46%), con un 83% de especificidad. El grado de correlacion entre los niveles de CA en el LBA y el tejido pulmonar fue de 0,633 (p = 0,002). Conclusiones El estudio del LBA ofrece una prueba objetiva de la exposicion a amianto. La buena correlacion observada entre los recuentos de CA en el LBA y en el tejido pulmonar indica la validez de ambas tecnicas para analizar el contenido de amianto.

Background: The immune microenvironment plays a role in tumorigenesis. Chronic Obstructive Pulmonary Disease (COPD) is an independent risk factor for lung cancer (LC). We hypothesized that immune profile characterized by T regulatory (Treg), natural killer (NK), and plasma cells, as well as interleukin (IL)-10 and interferon-gamma, may differ within tumors of LC patients with/without COPD. Methods: Treg (anti-CD3 and anti-forkhead boxP3 antibodies), NK (anti-NCR1 antibody), IgG (anti-CD138-IgG antibody), IgA (anti-CD138-IgA antibody) using immunohistochemistry, and both IL-10 and interferon-gamma (ELISA) were quantified in tumor and non-tumor specimens (thoracotomy for lung tumor resection) from 33 LC&ndash, COPD patients and 20 LC-only patients. Results: Immune profile in tumor versus non-tumor specimens: Treg cell counts significantly increased in tumors of both LC and LC&ndash, COPD patients, while in tumors of the latter group, IgG-secreting plasma cells significantly decreased and IL-10 increased. No significant differences were seen in levels of NK cells, IgA-secreting cells, IgA/IgG, or interferon-gamma. Immune profile in tumors of LC&ndash, COPD versus LC: No significant differences were observed in tumors between LC&ndash, COPD and LC patients for any study marker. Conclusions: Immune cell subtypes and cytokines are differentially expressed in lung tumors, and the presence of COPD elicited a decline in IgG-secreting plasma cell levels but not in other cell types.

With the identification of therapeutic targets for lung adenocarcinoma, it has become mandatory to distinguish it from other entities. Some cases remain classified as non-small cell lung carcinoma, not otherwise specified (NSCLC-NOS) with immunohistochemistry. Electron microscopy (EM) can be useful, allowing the identification of glandular differentiation. The aim of this study was to determine the complementary value of immunohistochemistry and EM.Forty-eight NSCLC-NOS cases were selected (PSMAR-Biobank, Barcelona, Spain). Immunohistochemistry (TTF-1, p40) was performed. Tissue was retrieved from paraffin blocks. Results were compared to the final diagnosis, derived from combination of light microscopy, immunohistochemistry, EM, molecular studies and resection specimen.Immunohistochemistry concurred with final diagnosis in 36 cases (75%, Kappa = 0.517). EM agreed with final diagnosis in 35 (72.9%, Kappa = 0.471). Immunohistochemistry had a sensitivity = 73%, specificity = 100%, positive predictive value (PPV) = 100% and negative predictive value (NPV) = 52.4% for adenocarcinoma. All adenocarcinoma cases not solved by immunohistochemistry (n = 10) were classified by EM, and vice versa. Data from EM were identical to those of immunohistochemistry: sensitivity = 73%, specificity = 100%, PPV = 100% and NPV = 52.4%. Combining both techniques, 47 cases were coincident with final diagnosis (97.9%, Kappa = 0.943).EM can provide valuable information in subtyping NSCLC-NOS, being particularly useful when immunohistochemistry is inconclusive. EM could be considered as a complementary tool for decision-making in NSCLC-NOS.

Context.— Social media sites are increasingly used for education, networking, and rapid dissemination of medical information, but their utility for facilitating research has remained largely untapped. Objective.— To describe in detail our experience using a social media platform (Twitter) for the successful initiation, coordination, and completion of an international, multi-institution pathology research study. Design.— Following a tweet describing a hitherto-unreported biopsy-related histologic finding in a mediastinal lymph node following endobronchial ultrasound–guided transbronchial needle aspiration, a tweet was posted to invite pathologists to participate in a validation study. Twitter's direct messaging feature was used to create a group to facilitate communication among participating pathologists. Contributing pathologists reviewed consecutive cases of mediastinal lymph node resection following endobronchial ultrasound–guided transbronchial needle aspiration and examined them specifically for biopsy site changes. Data spreadsheets containing deidentified data and digital photomicrographs of suspected biopsy site changes were submitted via an online file hosting service for central review by 5 pathologists from different institutions. Results.— A total of 24 pathologists from 14 institutions in 5 countries participated in the study within 143 days of study conception, and a total of 297 cases were collected and analyzed. The time interval between study conception and acceptance of the manuscript for publication was 346 days. Conclusions.— To our knowledge, this is the first time that a social media platform has been used to generate a research idea based on a tweet, recruit coinvestigators publicly, communicate with collaborating pathologists, and successfully complete a pathology study.

In 2011 the Spanish Society of Medical Oncology (SEOM) and the Spanish Society of Pathology (SEAP) started a joint project to establish guidelines on biomarker testing in patients with advanced non-small-cell lung cancer (NSCLC) based on current evidence. As this field is constantly evolving, these guidelines have been updated, previously in 2012 and 2015 and now in 2019. Current evidence suggests that the mandatory tests to conduct in all patients with advanced NSCLC are for EGFR and BRAF mutations, ALK and ROS1 rearrangements and PD-L1 expression. The growing need to study other emerging biomarkers has promoted the routine use of massive sequencing (next-generation sequencing, NGS). The coordination of every professional involved and the prioritisation of the most suitable tests and technologies for each case remains a challenge., SEAP and SEOM have received financial support for this project in the form of unrestricted grants from AstraZeneca, Novartis, Pfizer, Roche, Sysmex and Takeda. F. López-Rios, E. Conde and L. Paz-Ares thank the support of iLUNG-CM (B2017/BMD-3884).

Resumen Introduccion La tuberculosis (TBC) ganglionar mediastinica es rara y su presentacion intratoracica es un reto diagnostico. La puncion aspirativa transbronquial guiada por ultrasonografia endobronquial (PATb-USEB) podria ser util para su diagnostico, pero no disponemos de estudios que evaluen su eficacia en nuestro medio. Metodos Se analizaron retrospectivamente todos los pacientes con TBC ganglionar mediastinica aislada diagnosticados entre 2008 y 2014. Se identificaron los pacientes con diagnostico definitivo a quienes se les realizo PATb-USEB. Resultados Se identificaron 29 casos, siendo la PATb-USEB diagnostica en 28 pacientes (96,6%). Los hallazgos citologicos confirmaron granulomas en el 93% de los casos. El cultivo fue positivo en 14 (48,2%) y la PCR para Mycobacterium tuberculosis , en el 30% de los casos en los que se realizo. El 84% de los pacientes eran inmigrantes y al compararlos con la poblacion nativa se encontraron diferencias significativas en el estado inmunologico y el rendimiento del cultivo. No se registraron complicaciones derivadas del procedimiento. Conclusion La PATb-USEB es una tecnica de primera linea, efectiva y segura en el diagnostico de pacientes con sospecha de TBC mediastinica.

Immune profile of B and T cells and tertiary lymphoid structures (TLSs) may differ in tumors of lung cancer (LC) patients with/without chronic obstructive pulmonary disease (COPD), and may also influence patient survival. We sought to analyze: (1) TLSs, germinal centers (GCs), B and T cells, and (2) associations of the immune biomarkers with the patients&rsquo, 10-year overall survival (OS). TLSs (numbers and area), B [cluster of differentiation (CD) 20], and T (CD3), and GCs cells were identified in both tumor and non-tumor specimens (thoracotomy) from 90 LC-COPD patients and 43 LC-only patients. Ten-year OS was analyzed in the patients. Immune profile in tumors of LC-COPD versus LC: TLS numbers and areas significantly decreased in tumors of LC-COPD compared to LC patients. No significant differences were observed in tumors between LC-COPD and LC patients for B or T cells. Immune profile in tumors versus non-tumor specimens: TLS areas and B cells significantly increased, T cells significantly decreased in tumors of both LC and LC-COPD patients. Survival: in LC-COPD patients: greater area of TLSs and proportion of B cells were associated with longer survival rates. The immune tumor microenvironment differs in patients with underlying COPD and these different phenotypes may eventually impact the response to immunotherapy in patients with LC.

Background Detection of ALK and ROS1 gene rearrangements in non–small-cell lung cancer is required for directing patient care. Although fluorescence in situ hybridization (FISH) and immunohistochemistry have been established as gold standard methods, next-generation sequencing (NGS) platforms are called to be at least equally successful. Comparison of these methods for translation into daily use is currently under investigation. Patients and Methods Forty non–small-cell lung cancer paraffin-embedded samples with previous ALK (n = 33) and ROS1 (n = 7) FISH results were examined with the Oncomine Focus Assay and tested for ALK and ROS1 immunoreactivity. Clinical implications of concurrent molecular alterations and concordance between methods were evaluated. Results NGS was successful in 32 (80%) cases: 25 ALK and 7 ROS1. Few concomitant alterations were detected: 1 ALK rearranged case had an ALK p.L1196M-resistant mutation, 4 had CDK4, MYC, and/or ALK amplifications, and 1 ROS1 rearranged case showed a FGFR4 amplification. Comparison between techniques revealed 5 (16%) discordant cases that had lower progression-free survival than concordant cases: 7.6 (95% confidence interval, 2.2-13) versus 19.4 (95% confidence interval, 10.1-28.6). Remarkably, 4 of these cases had isolated 3' signal FISH pattern (P = .026). Conclusion Our data support that the identification of 3' isolated signal FISH pattern in ALK and ROS1 cases might suggest a false-positive result. NGS seems a reliable technique to assess ALK and ROS1 rearrangements, offering the advantage over immunohistochemistry of detecting other molecular alterations with potential therapeutic implications.

OBJECTIVE: We aimed to assess MET intratumoral heterogeneity and its potential impact on biomarker-based patient selection as well as potential surrogate biomarkers of MET activation. METHODS: Our study included 120 patients with non-squamous Non-small-cell Lung Cancer (nsNSCLC), of which 47 were incorporated in tissue microarrays (TMA). Four morphologically distinct tumor areas were selected to assess MET heterogeneity. MET positivity by immunohistochemistry (IHC) was defined as an above-median H-score and by +2/+3 staining intensity in >50% of tumor cells (Metmab criteria). MET FISH positivity was defined by MET/CEP7 ratio ≥ 2.0 and/or MET ≥ 5.0. MET staining pattern (cytoplasmic vs. membranous) and mesenchymal markers were investigated as surrogates of MET activation. RESULTS: Median MET H-score was 140 (range 0-400) and 47.8% of patients were MET positive by Metmab criteria. Eight cases (6.8%) were MET FISH positive and showed higher H-scores (p = 0.021). MET positivity by IHC changed in up to 40% of cases among different tumor areas, and MET amplification in 25-50%. Cytoplasmic MET staining and positivity for vimentin predicted poor survival (p = 0.042 and 0.047, respectively). CONCLUSIONS: MET status is highly heterogeneous among different nsNSCLC tumor areas, hindering adequate patient selection for MET-targeted therapies. MET cytoplasmic staining and vimentin might represent surrogate markers for MET activation. This work was supported by RD12/0036/0051 /FEDER, PI13/00140/FEDER, a grant from Fundacio Marato de TV3. Ref.666/C/2013 and 2014 SGR 740. the “Xarxa de Bancs de tumors sponsored by Pla Director d’Oncologia de Catalunya (XBTC).” JA is recipient of intensification programme ISCIII.

Resumen Introduccion La Anatomia Patologica sigue siendo desconocida, no solo para el publico en general, sino tambien para nuestros companeros de profesion. El uso generalizado de las redes sociales permite difundir nuestra labor. Por ello, se planteo generar un grupo de trabajo. Material y metodos Se emitio una convocatoria a los socios de la Sociedad Espanola de Anatomia Patologica a traves de su Vocalia de Comunicacion y Proyeccion Social titulada #IWantYouForSEAP, para conformar una red de trabajo en la red social Twitter. El periodo de reclutamiento fue de un mes (agosto del 2016). Se registro la actividad desarrollada durante el XXVIII Congreso de la SEAP-IAP de 2017 utilizando las herramientas analiticas Symplur y Tweet Binder. Resultados Se recibieron 32 solicitudes (29 patologos, 2 tecnicos de Anatomia Patologica, un administrativo), con una distribucion geografica dentro del territorio espanol que incluyo a participantes de 14 de las 17 comunidades autonomas, perteneciendo a 22 ciudades y 25 centros distintos. La actividad en relacion con el hashtag #SEAP2017V utilizado en el congreso incluyo a 685 participantes, 6.704 tweets y 8.837.435 impresiones; 28 de los 32 reclutados por la campana #IWantYouForSEAP participaron, contribuyendo con 2.410 tweets y generando 2.090.423 impresiones (el 36 y el 24% del total, respectivamente). Conclusiones Es posible promover y motivar el trabajo en grupo dentro de nuestra disciplina a traves de las redes sociales. Se trata de una fase inicial, pero los resultados experimentados son alentadores y existen grandes expectativas por parte de los participantes. Un uso adecuado de las redes sociales podria acercar la figura del patologo a la sociedad.

Background Chronic lung diseases such as chronic obstructive pulmonary disease (COPD) and epigenetic events underlie lung cancer (LC) development. The study objective was that lung tumor expression levels of specific microRNAs and their downstream biomarkers may be differentially regulated in patients with and without COPD. Methods In lung specimens (tumor and non-tumor), microRNAs known to be involved in lung tumorigenesis (miR-21, miR-200b, miR-126, miR-451, miR-210, miR-let7c, miR-30a-30p, miR-155 and miR-let7a, qRT-PCR), DNA methylation, and downstream biomarkers were determined (qRT-PCR and immunoblotting) in 40 patients with LC (prospective study, subdivided into LC-COPD and LC, N = 20/group). Results Expression of miR-21, miR-200b, miR-210, and miR-let7c and DNA methylation were greater in lung tumor specimens of LC-COPD than of LC patients. Expression of downstream markers PTEN, MARCKs, TPM-1, PDCD4, SPRY-2, ETS-1, ZEB-2, FGFRL-1, EFNA-3, and k-RAS together with P53 were selectively downregulated in tumor samples of LC-COPD patients. In these patients, tumor expression of miR-126 and miR-451 and that of the biomarkers PTEN, MARCKs, FGFRL-1, SNAIL-1, P63, and k-RAS were reduced. Conclusions Biomarkers of mechanisms involved in tumor growth, angiogenesis, migration, and apoptosis were differentially expressed in tumors of patients with underlying respiratory disease. These findings shed light into the underlying biology of the reported greater risk to develop LC seen in patients with chronic respiratory conditions. The presence of an underlying respiratory disease should be identified in all patients with LC as the differential biological profile may help determine tumor progression and the therapeutic response. Additionally, epigenetic events offer a niche for pharmacological therapeutic targets. Electronic supplementary material The online version of this article (10.1186/s13148-017-0437-0) contains supplementary material, which is available to authorized users.

Background The assessment of epidermal growth factor receptor (EGFR) mutations is crucial for the management of patients with lung adenocarcinoma. Circulating tumor DNA (ctDNA)-based assessment offers advantages over tumor as a minimally invasive method able to capture tumor heterogeneity. Patients and Methods Consecutive patients diagnosed with EGFR-mutant lung adenocarcinoma in tumor biopsy were included in this study. Plasma samples were obtained at different time points during the course of the disease. EGFR mutations in plasma were quantified using BEAMing (beads, emulsions, amplification, and magnetics) or digital PCR and were correlated with mutations in tumor and with radiologic response and progression. Results Two hundred twenty-one plasma samples from 33 patients were analyzed. EGFR mutations in plasma were detected in 83% of all patients and 100% of those with extrathoracic metastases. The dynamics of the EGFR mutation load predicted response in 93% and progression in 89% of cases well in advance of radiologic evaluation. Progression-free survival for patients in whom ctDNA was not detected in plasma during treatment was significantly longer than for those in whom ctDNA remained detectable (295 vs. 55 days; hazard ratio, 17.1; P Conclusion The detection of EGFR mutations in ctDNA showed good correlation with that in tumor biopsy and predicted tumor response and progression in most patients. The liquid biopsy for ctDNA-based assessment of EGFR mutations is a reliable technique for diagnosis and follow-up in patients with EGFR-mutant lung adenocarcinoma in routine clinical practice.

Although adenocarcinoma (ADC) is the most frequent lung cancer, its diagnosis is often late, when the local invasion is important and/or the metastases have already appeared. Therefore, the mortality at 5 years is still very high, ranging from 51% to 99%, depending on the stage. The implementation of different molecular techniques has allowed genomic studies even in relatively small histological samples such as obtained with non-invasive or minimally invasive techniques, facilitating a better phenotyping of lung ADC. Thus, current classification differentiates between preinvasive lesions (atypical adenomatous hyperplasia and in situ ADC), minimally invasive ADC (MIA) and invasive ADC. 'Field cancerization' is a concept that refers to progressive loco-regional changes occurring in tissues exposed to carcinogens, due to the interaction of the latter with a predisposing genetic background and an appropriate tissue microenvironment. Somatic genetic alterations, including mutations but also other changes, are necessary for oncogenesis, being especially frequent in lung ADC. Changes in the epidermal growth factor receptor (EGFR) gene, Kirsten rat sarcoma viral oncogene (KRAS), v-Raf murine sarcoma viral oncogene homolog B (BRAF), gene encoding neurofibromin (NF1), anaplastic lymphoma kinase (ALK) and ROS1 are the main genes that suffer alterations in the tumors of patients with ADC. Molecular profiling of these tumors allows more targeted treatments through two distinct strategies, genome-guided therapy and immunotherapy. The former, targets the aberrant pathways secondary to the genomic alteration, whereas the latter may be based on the administration of antibodies [such as those against cytotoxic T-lymphocyte antigen 4 (CTLA-4) or the programmed cell death ligand 1/protein 1 pathway (PD-L1/PD-1)] or the stimulation of the patient's own immune system to produce a specific response. These strategies are obtaining better results in selected ADC patients.

Pathology is a speciality that is often poorly understood, not only by the general public, but also by clinicians. However, the recent widespread use of social media provides an opportunity to increase the visibility and comprehension of our profession. A working group was formed to carry out this task.The members of the Spanish Society of Pathology were contacted through its Communication and Social Projection Subcommittee to engage in the campaign #IWantYouForSEAP, to form a network on Twitter. The recruitment period was one month (August, 2016). The resulting project, developed during the XXVIII Congress of the SEAP-IAP, was registered using the analytical tools Symplur and Tweet Binder.32 applications (29 pathologists, 2 histotechnicians, 1 administrative personnel) were received from all over Spain, including participants from 14 of the 17 Autonomous Regions, from 22 cities and 25 medical centres. The activity in relation to the hashtag #SEAP2017V used in the congress included 685 participants with 6704 tweets and 8,837,435 impressions. 28 of the 32 recruited by the #IWantYouForSEAP campaign participated, contributing with 2410 tweets, and generating 2,090,423 impressions (36% and 24% of the total, respectively).It is possible to promote and motivate teamwork within our discipline through social media networks. This preliminary experience of the use of social media networks in our scientific community has had encouraging results which have raised high expectations among participants. An appropriate use of social media networks could help to narrow the gap between pathologists and society.

Resumen Introduccion La broncoscopia guiada por fluoroscopia se utiliza para el diagnostico de las lesiones pulmonares perifericas (LPP), pero su rendimiento es muy variable en funcion de los estudios. La ultrasonografia endobronquial (USEB) radial podria incrementar el rendimiento diagnostico de la broncoscopia para estas lesiones. Objetivos Comparar el rendimiento diagnostico de la broncoscopia guiada por fluoroscopia y USEB radial con la broncoscopia guiada por fluoroscopia para el estudio de LPP. Metodos Se incluyeron de forma prospectiva todos los pacientes que se sometieron a broncoscopia con fluoroscopia para el estudio de LPP desde enero de 2009 a diciembre de 2012. Los 145 pacientes se aleatorizaron en 2 grupos: fluoroscopia y USEB radial (50 pacientes, 71,3 ± 8,2 anos) o fluoroscopia unicamente (95 pacientes, 68 ± 10,5 anos). El diametro medio de las lesiones fue de 41,97 ± 19,22 mm. Se tomaron muestras de cepillado bronquial citologico y biopsia transbronquial. Todas las exploraciones se realizaron bajo control fluoroscopico y con sedacion intravenosa. Para la USEB se utilizo un procesador ecografico equipado con una ultra-minisonda ecografica de 20 MHz que se introducia por una guia. Broncoscopista, citologo, protocolo de estudio, tecnicas y utillaje fueron los mismos durante todo el estudio. Resultados Ciento veintinueve (89%) pacientes presentaban patologia maligna. Se obtuvo el diagnostico por broncoscopia en 105 (72,4%) enfermos. En el grupo con fluoroscopia y USEB radial se diagnosticaron el 78% de los pacientes y en el grupo con solo fluoroscopia el 69,5% (n.s.). Sin embargo, para lesiones menores de 30 mm la fluoroscopia con USEB radial incrementaba significativamente el rendimiento diagnostico comparado con la fluoroscopia unicamente (90 vs. 52%; p = 0,05). Conclusiones La USEB radial asociada a fluoroscopia es una tecnica especialmente util para el diagnostico de las LPP de un tamano inferior a 30 mm.

Resumen Introduccion El estadio de la enfermedad es el factor pronostico mas importante en el cancer de pulmon, y una estadificacion optima es importante para determinar la mejor opcion terapeutica. La FDG-PET/TC ha resultado util en el estadio inicial del cancer de pulmon no microcitico (CPNM) pero los datos existentes continuan siendo insuficientes para definir su papel en otros estadios. Hipotesis La informacion aportada por la FDG-PET/TC tiene repercusiones en el enfoque terapeutico adoptado ante los pacientes con CPNM. Metodos Se realizo una revision retrospectiva de los pacientes a los que se practico una FDG-PET/TC en el proceso diagnostico del CPNM entre enero de 2008 y diciembre de 2010. Se obtuvieron datos relativos al estadio clinico antes y despues de la FDG-PET/TC asi como informacion sobre el cambio de la decision terapeutica como consecuencia de la informacion aportada por la FDG-PET/TC. Utilizando como patron de referencia el examen anatomopatologico, se calcularon los valores de sensibilidad, especificidad y valor predictivo positivo y negativo de la TC y la FDG-PET/TC. Resultados De los 522 pacientes con un diagnostico de CPNM, en 246 (47,1%) se realizo una FDG-PET/TC. En 85 casos (34,6%) la FDG-PET/TC comporto una migracion del estadio. El abordaje terapeutico se modifico en 60 pacientes (24,4% del total de las FDG-PET/TC realizadas), y ello permitio evitar una toracotomia futil en 13 casos (5,2%), e hizo posible un tratamiento con intencion curativa en 26 (10,5%). De los 90 pacientes (36,5%) clasificados en el estadio iii mediante la estadificacion de TC, la FDG-PET/TC modifico el abordaje terapeutico en 36 (40%). En los 133 casos (54%) con una evaluacion anatomopatologica de los ganglios linfaticos mediastinicos, la sensibilidad, la especificidad, el valor predictivo positivo y el valor predictivo negativo fueron de 0,57; 0,64; 0,48 y 0,72 con la TC; y de 0,68; 0,86; 0,75 y 0,81 con la FDG-PET/TC. Discusion Nuestros datos respaldan los de trabajos previos que indican que la FDG-PET/TC es esencial en el proceso de estadificacion, no solo en los pacientes con un CPNM potencialmente operable, sino tambien en los pacientes en estadio iii , tal como ponen de manifiesto nuestros datos.

Background MET amplifications present a potential therapeutic target in NSCLC and while FISH is conventionally used to asses it, there is no clinically defined cutoff. NGS is becoming routine in molecular diagnostics, and provides a means to assess MET amplification in the context of comprehensive genomic profiling. Here, we assess MET amplifications in NSCLC, in addition to TMB and concomitant driver mutations. Methods 222 NSCLC cases were screened, and 26 samples were selected based on increased MET gene copy number (GCN) detected by FISH (Cappuzzo score, MET GCN >5). Cases were reclassified to determine amplification versus high polysomy by University of Colorado Cancer Center (UCCC) FISH-criteria. Archival tissue samples were evaluated by PGDx elio™ tissue complete (assay under development; Personal Genome Diagnostics), a 500+ gene NGS panel. Results Eight patients were amplified by UCCC FISH-criteria (1 patient high-amplified (MET/CEP7 >5) and 7 patients intermediate-amplified (>2.2 to 3 fold cutoff) and were male smokers, with a median age of 57 years, with no concurrent driver alterations. One MET-amplified sample also had a concomitant exon 14 skipping mutation. The 3 discordant cases were highly heterogeneous by FISH with focal MET amplifications. Eighteen patients exhibited high copy number gain by FISH but were negative for MET amplifications according to UCCC FISH-criteria and NGS. Five of these cases had driver mutations (2 = KRAS exon 2 mutations and 3 = EGFR sensitizing mutations). TMB scores in NGS MET-positive were lower than NGS MET-negative cases: 5.9 vs. 12.5 mut/Mb exome equivalent. Conclusions NGS and FISH produced similar MET amplification status. The wide range of MET classifications via FISH, together with tumor heterogeneity and lack of clinically relevant thresholds, could contribute to discordance. In most cases negative for MET amplification by NGS, alternative driver mutations and/or higher TMB were identified. These results suggest that, while these technologies provide complementary value, further clinical data is needed to define clinically meaningful MET alterations. Legal entity responsible for the study The authors. Funding Has not received any funding. Disclosure E. Weingartner: Full / Part-time employment: Personal Genome Diagnostics. G. Cerqueira: Full / Part-time employment: Personal Genome Diagnostics. D. Nichol: Full / Part-time employment: Personal Genome Diagnostics. J. Simmons: Full / Part-time employment: Personal Genome Diagnostics. All other authors have declared no conflicts of interest.

Tumor recurrence is frequent and survival rates remain extremely low in lung adenocarcinoma (ADC). We hypothesize that carcinogenic factors will promote loco-regional modifications not only in the future tumor, but throughout the exposed lung. Objective: To analyze whether the most prevalent mutations observed in ADC can also be observed in the non-neoplastic lung tissue, as well as the short-term prognosis implications of this finding. Methods: Non-tumoral lung parenchyma specimens obtained during surgery from 47 patients with EGFR and/or KRAS abnormalities in their ADC tumors underwent similar genomic testing. Short-term outcomes were also recorded. Results: The same mutations were present in the tumor and the histologically normal tissue in 21.3% of patients (SM group). Although local recurrences were similar in both groups, distant metastases were more frequent in the former (60 vs. 5.4%, p &lt, 0.001). Moreover, SM patients showed lower time-to-progression (8.5 vs. 11.7 months, p &lt, 0.001) and disease-free survival (8.5 vs. 11.2 months, p &lt, 0.001). COX regression showed a higher risk of progression or death (DFS) in the SM group (HR 5.94, p &lt, 0.01]. Similar results were observed when adjusting for potential confounding variables. Conclusions: These results confirm that genetic changes are present in the apparently normal lung in many ADC patients, and this finding has prognostic implications.