Ignacio Escobar

Epigenetic Inactivation of microRNA-34b/c Predicts Poor Disease-Free Survival in Early-Stage Lung Adenocarcinoma

Purpose: The microRNA-34b/c (miR-34b/c) is considered a tumor suppressor in different tumor types and a transcriptional target of TP53. The main objectives of this study were to investigate the clinical implications of miR-34b/c methylation in patients with early-stage lung adenocarcinoma and to determine the functional role of miR-34b/c re-expression in lung adenocarcinoma cell lines. Experimental Design: Aberrant methylation and expression of miR-34b/c were assessed in 15 lung adenocarcinoma cell lines and a cohort of 140 early-stage lung adenocarcinoma. Lung adenocarcinoma cell lines were transfected with miR-34b/c and the effects upon cell proliferation, migration, invasion, and apoptosis were investigated. Results: Aberrant methylation of miR-34b/c was detected in 6 (40%) of 15 lung adenocarcinoma cell lines and 64 of 140 (46%) primary lung adenocarcinoma. Expression of miR-34b/c was significantly reduced in all methylated cell lines and primary tumors, especially with TP53 mutations. Patients with increased miR-34b/c methylation had significantly shorter disease-free and overall survival as compared to patients with unmethylated or low level of miR-34b/c methylation. Ectopic expression of miR-34b/c in lung adenocarcinoma cell lines decreased cell proliferation, migration, and invasion. Conclusions: Epigenetic inactivation of miR-34b/c by DNA methylation has independent prognostic value in patients with early-stage lung adenocarcinoma. Reexpression of miR-34b/c leads to a less aggressive phenotype in lung adenocarcinoma cell lines. Clin Cancer Res; 19(24); 6842–52. ©2013 AACR.

Lung fibrotic tenascin-C upregulation is associated with other extracellular matrix proteins and induced by TGFβ1

BackgroundIdiopathic pulmonary fibrosis (IPF) is a progressive parenchymal lung disease of unknown aetiology and poor prognosis, characterized by altered tissue repair and fibrosis. The extracellular matrix (ECM) is a critical component in regulating cellular homeostasis and appropriate wound healing. The aim of our study was to determine the expression profile of highlighted ECM proteins in IPF lungs.MethodsECM gene and protein expression was analyzed by cDNA microarrays, rt-PCR, immunohistochemistry and western-blot in lungs from idiopathic pulmonary fibrosis (IPF), hypersensitivity pneumonitis (HP), categorized as chronic (cHP) and subacute (saHP), and healthy lung tissue. Primary fibroblast cultures from normal subjects and fibrotic patients were studied to evaluate tenascin-C (TNC) synthesis.ResultsA total of 20 ECM proteins were upregulated and 6 proteins downregulated in IPF. TNC was almost undetected in normal lungs and significantly upregulated in fibrotic lungs (IPF and cHP) compared to saHP. Furthermore, it was located specifically in the fibroblastic foci areas of the fibrotic lung with a subepithelial gradient pattern. TNC levels were correlated with fibroblastic foci content in cHP lungs. Versican and fibronectin glycoproteins were associated with TNC, mainly in fibroblastic foci of fibrotic lungs. Fibroblasts from IPF patients constitutively synthesized higher levels of TNC than normal fibroblasts. TNC and α-sma was induced by TGF-β1 in both fibrotic and normal fibroblasts. TNC treatment of normal and fibrotic fibroblasts induced a non-significant increased α-sma mRNA.ConclusionsThe difference in ECM glycoprotein content in interstitial lung diseases could contribute to the development of lung fibrosis. The increase of TNC in interstitial areas of fibrotic activity could play a key role in the altered wound healing.

Quality study of a lung cancer committee: study of agreement between preoperative and pathological staging.

OBJECTIVE Accurate preoperative staging is essential to provide the best treatment for lung cancer. The objective of the present study was to determine agreement between preoperative and surgical-pathological staging and to analyse the impact of any disparity on treatment. METHODS This is a descriptive study of a series of 176 lung cancer cases treated by surgery between 2005 and 2007. Preoperative staging was based on clinical information and computed tomography (CT), positron emission tomography (PET), PET-CT, bronchoscopy and mediastinoscopy. In all cases, surgical-pathological staging was based on the analysis of surgical samples and the findings during surgery. Both preoperative and pathological stage determination were based on the TNM (tumour, node, metastasis) classification established in 1997. Concordance was measured by calculating agreement rates and the kappa value. RESULTS Preoperative and surgical-pathological staging agreed in 102 cases, an agreement rate of 58% and kappa value of 0.54 (95% confidence interval (CI) 0.44-0.63). The highest kappa value (0.68, 95% CI 0.53-0.82) was obtained in stage IA patients. Patients who underwent PET or PET-CT had a better kappa index (0.56, 95% CI 0.45-0.67, vs 0.39, 95% CI 0.21-0.56). Surgical-pathological staging validated surgery in 145 cases (82%), while 21 (12%) were revised to stage IIIA N2 and 10 (6%) to non-surgical stages. CONCLUSIONS Global agreement between preoperative and surgical-pathological staging was moderate. The best agreement was found in stages IV and IA.

An assessment of plantar hyperhidrosis after endoscopic thoracic sympathicolysis

BACKGROUND Endoscopic bilateral thoracic sympathicolysis (EBTS) is an effective and minimally invasive procedure used for patients with primary hyperhidrosis. The purpose of this study was to examine plantar hyperhidrosis before and after EBTS. METHODS A total of 198 patients with primary hyperhidrosis underwent 396 thoracoscopic sympathicolysis of ganglia T2-T3 in a prospective study. All completed a preoperative questionnaire, followed by a second questionnaire 12 months after the operation. The questionnaires evaluated sweating in the different body areas. Only the zones of anhydrosis were considered in delimiting the cutaneous expression of sympathetic ganglia T2-T3. RESULTS Redistribution of perspiration as reported by the patients comprised significant reductions in palmar and axillary hyperhidrosis, and an increase in the zone of the trunk and popliteal region. The incidence of plantar anhydrosis and plantar hypohidrosis was 30.3% and 20.7%, respectively (p < 0.001). CONCLUSIONS EBTS is followed by redistribution of body perspiration, with, and important, plantar anhydrosis and hypohidrosis. Although EBTS is the standard treatment for palmar primary hyperhidrosis, we must continue studying baseline sympathetic activity in patients affected by primary hyperhidrosis and the neuroanatomy of the sympathetic system to understand the redistribution of sweating and decrease of hyperhidrosis in the zones regulated by mental or emotional stimuli.

Localized (solitary) fibrous tumors of the pleura: an analysis of 15 patients.

BACKGROUND Localized fibrous tumors of the pleura (LFTPs) are rare neoplasms, which are considered to originate from submesothelial connective tissue. The aim of this article is to present 15 new cases because of their different clinical behaviors and to discuss the treatment of choice of such neoplasms. METHODS The records of 15 consecutive patients with LFTP operated at our Institution between 1995 and 2006 were retrospectively reviewed. Diagnostic procedures, clinical courses, and outcomes of these patients were studied. Total excision through a thoracotomy was performed in all patients. Neoplasms were considered to be malignant if one or more of the following histologic features were present: increasing mitotic activity; high cellularity with crowding and overlapping of nuclei; necrosis; and pleomorphism. RESULTS No operative mortality was reported. The mean follow-up time was 76 months. Malignant transformation was seen in 1 patient 26 months after resection of a benign tumor. Six cases were pathologically considered to be malignant: 2 patients developed local recurrence. One of these underwent redo-surgery and required pneumonectomy; in the other one surgery is not indicated because at the time of diagnosis the patient was 85 years. Currently, all patients are alive and 13 disease-free. CONCLUSIONS For histologically benign tumors, because of the risk of recurrence and malignant transformation, complete surgical resection is indicated and long-term follow-up is recommended in all patients. For malignant cases, complete surgical resection may be insufficient for the cure: further study should be performed to identify reliable prognostic factors to indicate and evaluate the effectiveness of systemic treatment.

Hiperhidrosis primaria. Situación actual de la cirugía del simpático

Primary hyperhidrosis-PH is an excessive sweating without known etiology. The PH is more frequent in women and in palms, soles and axillae. Medical treatment is not effective. The objective of the surgery is to remove or to disconnect sympathetic ganglia T2 (craniofacial PH or facial blushing), T3 (palmar PH) and T3-T4 (axillary PH). The surgical techniques are mainly resection/transection, ablation with electrocoagulation, sympathetic block by clipping and radiofrequency. Anhidrosis is achieved in 95% of the patients. The overall rate of complications is less than 5% and these are minor complications. The most important unwanted effect is reflex sweating, presented in 48% of the patients. Reflex sweating is more frequent in back, thorax and abdomen and it appears independently of the surgical technique. Ninety percent of the patients are very satisfied after surgery. Nowadays, thoracic sympathetic surgery is the gold standard for primary hyperhidrosis.

Increased AGE-RAGE ratio in idiopathic pulmonary fibrosis

BackgroundThe abnormal epithelial-mesenchymal restorative capacity in idiopathic pulmonary fibrosis (IPF) has been recently associated with an accelerated aging process as a key point for the altered wound healing. The advanced glycation end-products (AGEs) are the consequence of non-enzymatic reactions between lipid and protein with several oxidants in the aging process. The receptor for AGEs (RAGEs) has been implicated in the lung fibrotic process and the alveolar homeostasis. However, this AGE-RAGE aging pathway has been under-explored in IPF.MethodsLung samples from 16 IPF and 9 control patients were obtained through surgical lung biopsy. Differences in AGEs and RAGE expression between both groups were evaluated by RT-PCR, Western blot and immunohistochemistry. The effect of AGEs on cell viability of primary lung fibrotic fibroblasts and alveolar epithelial cells was assessed. Cell transformation of fibrotic fibroblasts cultured into glycated matrices was evaluated in different experimental conditions.ResultsOur study demonstrates an increase of AGEs together with a decrease of RAGEs in IPF lungs, compared with control samples. Two specific AGEs involved in aging, pentosidine and Nε-Carboxymethyl lysine, were significantly increased in IPF samples. The immunohistochemistry identified higher staining of AGEs related to extracellular matrix (ECM) proteins and the apical surface of the alveolar epithelial cells (AECs) surrounding fibroblast foci in fibrotic lungs. On the other hand, RAGE location was present at the cell membrane of AECs in control lungs, while it was almost missing in pulmonary fibrotic tissue. In addition, in vitro cultures showed that the effect of AGEs on cell viability was different for AECs and fibrotic fibroblasts. AGEs decreased cell viability in AECs, even at low concentration, while fibroblast viability was less affected. Furthermore, fibroblast to myofibroblast transformation could be enhanced by ECM glycation.ConclusionsAll of these findings suggest a possible role of the increased ratio AGEs-RAGEs in IPF, which could be a relevant accelerating aging tissue reaction in the abnormal wound healing of the lung fibrotic process.

Vascular disease in COPD: systemic and pulmonary expression of PARC (Pulmonary and Activation-Regulated Chemokine)

Introduction The role of Pulmonary and Activation-Regulated Chemokine (PARC) in the physiopathology of Chronic Obstructive Pulmonary Disease (COPD) is not fully understood. The aim of the present study is to analyze the expression of PARC in lung tissue and its relationship with the vascular remodeling of the systemic and pulmonary arteries of COPD subjects. Methods To achieve this objective, protein and gene expression experiments, together with ELISA assays, were performed on the lung tissue, intercostal arteries and serum samples from COPD patients, non-obstructed smokers (NOS) and never-smokers (NS). Results A total of 57 subjects were included in the analysis (23 COPD, 18 NOS and 16 NS). In the comparisons between groups, a significantly increased lung protein expression of PARC was observed in the COPD group compared to the NOS group (1.96±0.22 vs. 1.29±0.27, P-adjusted = 0.038). PARC was located predominantly in the smooth muscle cells of the remodeled pulmonary muscular arteries and the macrophage-rich area of the alveolar parenchyma. No differences were detected in PARC gene expression analyses. The protein content of PARC in the intercostal arteries were similar between groups, though little remodeling was observed in these arteries. Circulating levels of PARC were numerically higher in patients with COPD compared to NOS and NS. Conclusion The results of the present study suggest an increased lung protein expression of PARC in COPD subjects. This protein was mainly localized in the smooth muscle cells of the pulmonary muscular arteries and was associated with the severity of intimal thickening, indicating its possible role in this remodeling process.

Primary hyperhidrosis. Current status of surgical treatment

Abstract Primary hyperhidrosis-PH is an excessive sweating without known etiology. The PH is more frequent in women and in palms, soles and axillae. Medical treatment is not effective. The objective of the surgery is to remove or to disconnect sympathetic ganglia T2 (craniofacial PH or facial blushing), T3 (palmar PH) and T3-T4 (axillary PH). The surgical techniques are mainly resection/transection, ablation with electrocoagulation, sympathetic block by clipping and radiofrequency. Anhidrosis is achieved in 95% of the patients. The overall rate of complications is less than 5% and these are minor complications. The most important unwanted effect is reflex sweating, presented in 48% of the patients. Reflex sweating is more frequent in back, thorax and abdomen and it appears independently of the surgical technique. Ninety percent of the patients are very satisfied after surgery. Nowadays, thoracic sympathetic surgery is the gold standard for primary hyperhidrosis.

Incidence, predictors and prognostic significance of thromboembolic disease in patients with advanced ALK-rearranged non-small cell lung cancer

Thromboembolic disease is fairly common in patients with lung cancer [1–3]. This incidence seems to be higher in patients with lung adenocarcinomas [4], with approximately 15% of those with advanced stage disease developing venous thromboembolisms (VTE) during the whole course of their disease [5–7]. Pulmonary adenocarcinomas are a heterogeneous group of diseases that can be stratified according to the presence of major oncogenic driver alterations. Anaplastic lymphoma kinase (ALK) rearrangements are detected in approximately 4% of these cases [8]. Isolated reports have suggested that patients bearing ALK-rearranged tumours might have a higher than expected incidence of thromboembolisms [9, 10]. In the present study, we have analysed the incidence, predictors and prognostic significance of thromboembolic events in a large, multi-institutional and homogeneous cohort of advanced stage patients with ALK-rearranged lung cancers from Spain and Portugal. Our primary objective was to estimate the incidence of thromboembolic events and their association with overall survival in these patients. High incidence and prognostic relevance of thromboembolic disease in patients with ALK-rearranged NSCLCs http://ow.ly/DEZr30j6kC8