Laureano Molins

miR-141 and miR-200c as Markers of Overall Survival in Early Stage Non-Small Cell Lung Cancer Adenocarcinoma

Background Several treatments in non-small cell lung cancer (NSCLC) are histology-dependent, and the need for histology-related markers is increasing. MicroRNAs (miRNAs) are promising molecular markers in multiple cancers and show differences in expression depending on histological subtype. The miRNA family miR-200 has been associated with the regulation of epithelial-mesenchymal (EMT)/mesenchymal-epithelial transition (MET). EMT involves profound phenotypic changes that include the loss of cell-cell adhesion, the loss of cell polarity, and the acquisition of migratory and invasive properties that facilitates metastasis. A dual role for the miR-200 family in the prognosis of several tumors has been related to tumor cell origin. However, the prognostic role and function of miR-200 family in early-stage NSCLC adenocarcinoma and squamous cell carcinoma (SCC) have not been well established. Methods miRNA expression was determined using TaqMan assays in 155 tumors from resected NSCLC patients. Functional studies were conducted in three NSCLC cell lines: H23, A-549 and HCC-44. Results High miR-200c expression was associated with shorter overall survival (OS) in the entire cohort (p = 0.024). High miR-200c (p = 0.0004) and miR-141 (p = 0.009) expression correlated with shorter OS in adenocarcinoma – but not in SCC. In the multivariate analysis, a risk score based on miR-141 and miR-200c expression emerged as an independent prognostic factor for OS in the entire cohort (OR, 2.787; p = 0.033) and in adenocarcinoma patients (OR, 10.649; p = 0.002). Functional analyses showed that miR-200c, was related to mesenchymal-epithelial transition (MET) and affected cell migration and E-cadherin levels, while overexpression of miR-141 reduced KLF6 protein levels and produced an increase of secretion of VEGFA in vitro (H23, p = 0.04; A-549, p = 0.03; HCC-44, p = 0.02) and was associated with higher blood microvessel density in patient tumor samples (p<0.001). Conclusion High miR-141 and miR-200c expression are associated with shorter OS in NSCLC patients with adenocarcinoma through MET and angiogenesis.

Low miR-145 and high miR-367 are associated with unfavourable prognosis in resected nonsmall cell lung cancer

The transcription factors SRY-related HMG box (SOX)2 and octamer-binding transcription factor (OCT)4 regulate the expression of the miR-302–367 cluster. miR-145 regulates SOX2 and OCT4 translation and p53 regulates miR-145 expression. We analysed the expression of the miR-302–367 cluster and miR-145 and the mutational status of p53 in resected nonsmall cell lung cancer (NSCLC) patients and correlated results with time to relapse (TTR). Tumour and paired normal tissue samples were obtained from 70 NSCLC patients. MicroRNA expression was assessed with TaqMan MicroRNA Assays. p53 exons 5 to 8 were sequenced. miR-145 was downregulated (p<0.0001) and miR-367 was upregulated (p<0.0001) in tumour compared with normal tissue. Mean TTR was 18.4 months for patients with low miR-145 levels and 28.2 months for those with high levels (p=0.015). Mean TTR was 29.1 months for patients with low miR-367 levels and 23.4 months for those with high levels (p=0.048). TTR was shorter for patients with both unfavourable variables (p=0.009). Low miR-145 expression (p=0.049), the combination of unfavourable microRNA levels (p=0.02) and the combination of low miR-145 with p53 mutations (p=0.011) were independent markers of shorter TTR. In conclusion, miR-145 and miR-367 expression could be novel markers for relapse in surgically treated NSCLC. p53 may play a role in modulating miR-145 expression in NSCLC.

False-negative rate after positron emission tomography/computer tomography scan for mediastinal staging in cI stage non-small-cell lung cancer

OBJECTIVES To assess the false-negative (FN) rate of positron emission tomography (PET)-chest computed tomography (CT) scan in clinical non-central cIA and cIB non-small-cell lung cancer (NSCLC) for mediastinal staging. METHODS Between January 2007 and December 2010, 402 patients with potentially operable NSCLC were assessed by thoracic CT scan and 18-fluoro-2-deoxy-d-glucose PET-CT for mediastinal staging and to detect extrathoracic metastases, of which 153 surgically treated patients (79 cIA and 74 cIB cases) were prospectively included in the study. Central tumours were excluded on the basis of CT scan criteria, defined as contact with the intrapulmonary main bronchi, pulmonary artery, pulmonary veins or the origin of the first segmental branches. CT scan was considered negative if lymph nodes were <1 cm at the smaller diameter. 18FDG PET-CT was considered negative when the high maximum standard uptake value (SUVmax) was <2.5. Non-invasive surgical staging was carried out in this group, and curative resection plus systematic mediastinal dissection was performed except in the event of unexpected oncological contraindication. RESULTS Composite non-invasive staging (CT scan, PET-CT) showed a negative predictive value (NPV) of 92% (CI 83.6-96.8) in the cIA group and 85% (CI 74-92) in the cIB group. There were 6 of 79 (7.6%) false-negatives (FNs) in cIA and 11 of 74 (14.8%) in cIB. Multilevel pN2 were detected in four cases, all of them in the cIB group. The most frequently involved N2 was subcarinal (two cases) in cIA and right lower paratracheal (R4) and seven (five cases) in cIB. Occult (pN2) lymph nodes were more frequent in tumour sizes≥5 cm (pT2b, nine cases, four FNs, P=0.03), pN1, adenocarcinoma [excluding minimally invasive adenocarcinoma (MIA) and lepidic predominant growth (LPA)] (P=0.029) and female patients, but no other risk factors for mediastinal metastases were identified (age, clinical stage, tumour location, central or peripheral, P>0.05). Multilevel pN2 was significantly more frequent in the cIB group (P<0.03). In pT≤1 cm (T1a), NPV was significantly better (NPV=100%, P<0.05) than the other subgroups studied (IA>1 cm and IB). CONCLUSIONS Composite results for non-invasive mediastinal staging (CT scan, PET-CT) showed 11% of FNs in cI stage (7.6% in non-central cIA and 14.8% in cIB). In tumours≤1 cm, NPV makes surgical staging unnecessary. In women with adenocarcinoma and non-central cIB, however, the high FN rate makes invasive staging necessary, particularly in pT2b to decrease the incidence of unexpected pN2 in thoracotomy.

Experiencia piloto de benchmarking en cirugía torácica: comparación de la casuística e indicadores de calidad en resección pulmonar

Objetivo En el presente articulo se describen los metodos y las conclusiones del primer estudio espanol de benchmarking en cirugia toracica. Los objetivos propuestos fueron: describir la casuistica de reseccion pulmonar desarrollada en 9 hospitales espanoles, comparar indicadores de calidad entre los 9 centros participantes e identificar y proponer areas de mejora comunes para los procesos de reseccion pulmonar. Metodos Se utilizo como fuente de informacion el conjunto minimo basico de datos de los anos 2002 y 2003 de los procesos de lobectomia o neumonectomia. Los indicadores de resultados seleccionados fueron: mortalidad hospitalaria, morbilidad, estancia y readmisiones urgentes en los 30 dias siguientes al alta, ajustadas por complejidad de los casos. Una vez presentados los resultados entre los participantes, se identificaron las unidades con mejores resultados y se discutieron diversas propuestas de mejora. Resultados Se ha estudiado un total de 1.666 procedimientos (1.276 lobectomias y 390 neumonectomias). Se detectaron diferencias en estancia media, mortalidad, tasa de readmisiones y morbilidad, que permitieron identificar unidades, de complejidad equiparable o superior, con baja mortalidad y estancia. Sin embargo, en estas unidades se apreciaron tasas de morbilidad y readmision mas elevadas. Conclusiones Se propusieron medidas encaminadas a registrar todos los diagnosticos relevantes en los informes de alta, disminuir las estancias inadecuadas y estandarizar los procedimientos que permitiran en el futuro establecer criterios fiables para mejorar la calidad de los procesos de reseccion pulmonar.

Comparative analysis of analgesic quality in the postoperative of thoracotomy: paravertebral block with bupivacaine 0.5% vs ropivacaine 0.2%.

OBJECTIVES Paravertebral block is an effective alternative to epidural analgesia in the management of post-thoracotomy pain, however, there are no established guidelines regarding what is the most suitable strategy when varying drugs and dosages between different groups. Our objective was to evaluate the effectiveness of paravertebral block comparing the most frequently employed drugs in this procedure (bupivacaine vs ropivacaine). METHODS Prospective randomized study of 70 patients submitted to thoracotomy. Patients were divided in two independent groups (anterior thoracotomy (AT) and posterolateral thoracotomy (PT)). At the end of surgery a catheter was inserted under direct vision in the thoracic paravertebral space at the level of incision. In each group (AT or PT) patients were randomized to receive a bolus of 15 ml of bupivacaine 0.5% or 20 ml of ropivacaine 0.2% before closing the thoracotomy. They postoperatively received 10 ml of bupivacaine or 15 ml of ropivacaine every 6h combined with methamizol (every 6h). Subcutaneous meperidine was employed as rescue drug. The level of pain was measured with the visual analogic scale (VAS) at 1, 6, 24, 48 and 72 h after surgery. The need of meperidine as rescue drug and secondary effects was also recorded. RESULTS We did not register secondary effects in relation to the paravertebral catheter (paravertebral or cutaneous bleeding or hematoma, respiratory depression (respiratory rate <8 breaths per minute), cardiotoxicity, confusion, sedation, urinary retention, nausea, vomiting and pruritus). Eleven patients (16%) needed meperidine as rescue drug (six with ropivacaine and five with bupivacaine). Mean VAS values were the following: all the cases (n=70): 5.2+/-2.1, AT (n=38): 4.5+/-2.1, PT (n=32): 5.9+/-1.7, bupivacaine (n=35): 4.9+/-2.1, ropivacaine (n=35): 5.4+/-1.9, AT with bupivacaine (n=19): 4.2+/-2.2, AT with ropivacaine (n=19): 4.9+/-2.0, PT with bupivacaine (n=16): 5.7+/-1.6, PT with ropivacaine (n=16): 6.0+/-1.7. CONCLUSIONS Post-thoracotomy analgesia combining paravertebral catheter and a nonsteroidal anti-inflammatory drug is a safe and effective practice, VAS values are acceptable (only 16% of patients required meperidine as rescue). It prevents the risk of side effects related to epidural analgesia. Patients submitted to AT experienced less pain than those with PT (4.5 vs 5.9, p<0.01). Bupivacaine got slightly better VAS values than ropivacaine (4.9 vs 5.4 p<0.05). Higher doses and volumes of local anesthetic could be used to obtain better VAS values.

YKT6 expression, exosome release, and survival in non-small cell lung cancer

Background Cancer-derived exosomes are involved in metastasis. YKT6 is a SNARE protein that participates in the regulation of exosome production and release, but its role in non-small cell lung cancer (NSCLC) has not been examined. Materials and Methods Ultracentrifugation-purified exosomes from the A549 cell line were studied by CRYO-TEM, nanoparticle tracking analysis and western blot (TSG101 marker). YKT6 was inhibited using a DsiRNA and selected pre-microRNAs. MicroRNAs targeting YKT6 were validated by Renilla/Luciferase assay and western blot. YKT6 expression and its prognostic impact were analyzed in 98 tissue specimens from resected NSCLC patients. Results Membranous nanosized vesicles (mode size: 128nm) with TSG101 protein were purified from A549 cells. YKT6 inhibition reduced exosome release by 80.9%. We validated miR-134 and miR-135b as miRNAs targeting YKT6, and transfection with the pre-miRNAs also produced a significant reduction in exosome release. The analysis of YKT6 in tumor samples showed that patients with high levels had shorter disease-free and overall survival. Conclusions YKT6 is a key molecule in the regulation of exosome release in lung cancer cells and is in turn precisely regulated by miR-134 and miR-135b. Moreover, YKT6 levels impact prognosis of resected NSCLC patients.

Assessment of a Combined Panel of Six Serum Tumor Markers for Lung Cancer

RATIONALE We have previously identified six serum tumor markers (TMs) (carcinoembryonic antigen, carbohydrate antigen 15.3, squamous cell carcinoma-associated antigen, cytokeratin-19 fragment, neuron-specific enolase, and pro-gastrin-releasing peptide) related to the presence of lung cancer (LC). OBJECTIVES To validate their individual performance in an independent cohort, and to explore if their combined assessment (≥1 abnormal TM value) is a more accurate marker for LC presence. METHODS We determined these six TMs in 3,144 consecutive individuals referred to our institution by their primary care physician because of the clinical suspicion of LC. MEASUREMENTS AND MAIN RESULTS LC was excluded in 1,316 individuals and confirmed in 1,828 patients (1,563 with non-small cell LC and 265 with small cell LC). This study validated the previously reported performance of each individual TM. We also showed that their combined assessment (≥1 abnormal TM) had a better sensitivity, specificity, negative predictive value, and positive predictive value (88.5, 82, 83.7, and 87.3%, respectively) than each TM considered individually and that it increased the diagnostic performance (area under the curve) of a clinical model that included tumor size, age, and smoking status. In patients with radiographic nodules less than 3 cm, the negative predictive value of the TM panel was 71.8%, hence providing some support for a more conservative diagnostic approach. Finally we identified two TMs (neuron-specific enolase and pro-gastrin-releasing peptide) that differentiate the risk of non-small cell LC from that of small cell LC. CONCLUSIONS The combined assessment of a panel of six serum TMs is a more accurate marker for LC presence than these same TMs considered individually. The potential of these TMs in the diagnostic and screening settings deserves further research.

Effectiveness of sympathetic block by clipping in the treatment of hyperhidrosis and facial blushing.

Main cause of dissatisfaction after videothoracoscopic (VATS) sympathectomy in the treatment of hyperhidrosis (HH) and facial blushing (FB) is compensatory sweating (CS). Sympathetic nerve (SN) clipping permits to revert the block effect by removing the clips in case of an intolerable CS. We present our experience with this technique. Sixty-one patients diagnosed for HH and/or FB were operated on VATS SN clipping. Clipping levels varied in function for the sort of disease: FB: T2, palmar HH: T3, axillary HH: T4, palmar-axillary HH: T3-4. Results were evaluated 1 week, 3 months and 12 months after surgery. Fifty-eight of the 61 patients showed improvement of their symptoms (95%). There were minor complications in three patients (5.5%). CS was seen in 38/61 (62.2%), being labelled as mild in 33/61 (54%) and severe in 5/61 patients (8.2%), no patient qualified it as intolerable and it was not necessary to remove the clips for CS in any case. SN clipping by VATS is a safe and effective procedure for the management of FB and palmar/axillary HH. Levels of CS are similar or lesser than with the standard sympathicotomy and, if necessary, this technique allows to revert the sympathetic block removing the clips.

Beneficios del uso de dispositivos digitales para medir la fuga aérea después de una resección pulmonar: estudio prospectivo y comparativo

INTRODUCTION Persistent air leaks represent the most common pulmonary complication after elective lung resection. Since there are insufficient data in the literature regarding variability in the withdrawal of postoperative pleural drainages, we have designed a prospective, consecutive and comparative study to evaluate if the use of digital devices (Thopaz and DigiVent) to measure postoperative air leak compared to a Pleur-Evac varies on deciding when to withdraw chest tubes after lung resection. METHODS A prospective, consecutive and comparative trial was conducted in 75 patients who underwent elective pulmonary resection for non small cell lung cancer. This study compared two digitals devices with the current analogue version in 75 patients. The digital and analogue groups had 26, 24, and 25 patients, respectively. RESULTS Clinical population data were not statistically different between the groups. The withdrawal of the chest tube was Thopaz, 2.4 days; Digivent, 3.3 days and PleurEvac, 4.5 days. Patients and nurses were subjectively more comfortable with digital devices. Surgeons obtained more objective information with digital devices. The safety mechanism of the Thopaz was also subjectively better, and one patient was discharged home without complications after one week. CONCLUSIONS The digital and continuous measurement of air leak instead of the currently used static analogue systems reduced the chest tube withdrawal and hospital stay by more accurately and reproducibly measuring air leak. Intrapleural pressure curves from the Digivent may also help predict the optimal chest tube setting for each patient. The Thopaz alarm mechanism is very useful to prevent deficiencies in the mechanism and do not required wall suction.

The benefits of digital air leak assessment after pulmonary resection: Prospective and comparative study

Abstract Introduction Persistent air leaks represent the most common pulmonary complication after elective lung resection. Since there are insufficient data in the literature regarding variability in the withdrawal of postoperative pleural drainages, we have designed a prospective, consecutive and comparative study to evaluate if the use of digital devices (Thopaz and DigiVent) to measure postoperative air leak compared to a Pleur-Evac varies on deciding when to withdraw chest tubes after lung resection. Methods A prospective, consecutive and comparative trial was conducted in 75 patients who underwent elective pulmonary resection for non small cell lung cancer. This study compared two digitals devices with the current analogue version in 75 patients. The digital and analogue groups had 26, 24, and 25 patients, respectively. Results Clinical population data were not statistically different between the groups. The withdrawal of the chest tube was Thopaz, 2.4 days; Digivent, 3.3 days and PleurEvac, 4.5 days. Patients and nurses were subjectively more comfortable with digital devices. Surgeons obtained more objective information with digital devices. The safety mechanism of the Thopaz was also subjectively better, and one patient was discharged home without complications after one week. Conclusions The digital and continuous measurement of air leak instead of the currently used static analogue systems reduced the chest tube withdrawal and hospital stay by more accurately and reproducibly measuring air leak. Intrapleural pressure curves from the Digivent may also help predict the optimal chest tube setting for each patient. The Thopaz alarm mechanism is very useful to prevent deficiencies in the mechanism and do not required wall suction.