Yuichi Sakairi

EML4-ALK Fusion Gene Assessment Using Metastatic Lymph Node Samples Obtained by Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration

Purpose: Anaplastic lymphoma kinase (ALK) fusion genes represent novel oncogenes for non–small cell lung cancers (NSCLC). Several ALK inhibitors have been developed, and are now being evaluated in ALK-positive NSCLC. The feasibility of detecting ALK fusion genes in samples obtained by endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) was determined. The clinicopathologic characteristics of ALK-positive lung cancer were also analyzed. Experimental Design: From April 2008 to July 2009, NSCLC cases with hilar/mediastinal lymph node metastases detected by EBUS-TBNA were enrolled. Positive expression of ALK fusion protein was determined using immunohistochemistry, and ALK gene rearrangements were further examined to verify the translocation between ALK and partner genes using fluorescent in situ hybridization and reverse transcription-PCR. Direct sequencing of PCR products was performed to identify ALK fusion variants. Results: One hundred and nine cases were eligible for the analysis using re-sliced samples. Screening of these specimens with immunohistochemistry revealed ALK positivity in seven cases (6.4%), all of which possessed echinoderm microtubule–associated protein-like 4–ALK fusion genes as detected by fluorescent in situ hybridization and reverse transcription-PCR. All ALK-positive cases had an adenocarcinoma histology and possessed no EGFR mutations. Compared with ALK-negative cases, ALK-positive cases were more likely to have smaller primary tumors (P < 0.05), to occur at a younger age (<60 years; P < 0.05), and to occur in never/light smokers (smoking index < 400; P < 0.01). Mucin production was frequently observed in ALK-positive adenocarcinomas (29.4%; P < 0.01). Conclusions: EBUS-TBNA is a practical and feasible method for obtaining tissue from mediastinal and hilar lymph nodes that can be subjected to multimodal analysis of ALK fusion genes in NSCLC. Clin Cancer Res; 16(20); 4938–45. ©2010 AACR.

Rapid On-Site Cytologic Evaluation During Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration for Nodal Staging in Patients With Lung Cancer

BACKGROUND The utility of rapid on-site evaluation (ROSE) during endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) for lymph node staging in lung cancer is still controversial. The aim of this study was to assess the role of ROSE during EBUS-TBNA and the interpretation of its results. METHODS We performed a retrospective chart review of patients with suspected or diagnosed lung cancer who underwent EBUS-TBNA for lymph node staging. The slides were air-dried and Diff-Quik (American Scientific Products, McGaw Park, IL) staining was used for ROSE. Additional smears were prepared for Papanicolaou staining and any remaining sample was placed in 10% formalin for histologic evaluation. The results of ROSE were compared with the results of the final pathologic diagnosis. RESULTS EBUS-TBNA was performed in 438 patients on 965 lymph nodes. Eighty-four lymph nodes (8.7%) were determined insufficient for definitive diagnosis by final cytologic evaluation. However 45 of the 84 lymph nodes were able to be diagnosed by histologic examination. The non-diagnostic sampling rate was 4.0%. There were no false-positive results on ROSE; however 25 cases (5.7%) were falsely evaluated as negative on ROSE. The concordance rate for staging between ROSE and final pathologic diagnosis was 94.3%. The sensitivity, specificity, negative predictive value, and diagnostic accuracy rate of EBUS-TBNA for correct lymph node staging was 96.5%, 100%, 89.8%, and 98.2%, respectively. CONCLUSIONS ROSE during EBUS-TBNA for material adequacy showed a low rate of non-diagnostic sampling. There was a high agreement between the on-site and final pathologic evaluation during EBUS-TBNA; however immediate diagnosis should be approached with caution.

A Solitary Metastatic Lung Tumor from Thyroid Papillary Carcinoma Diagnosed by Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration (EBUS-TBNA): Report of a Case

A solitary metastatic tumor from thyroid papillary carcinoma is rarely diagnosed preoperatively. A 61-year-old woman was referred to us for investigation of a solitary nodule in the right lung field on a chest X-ray, several years after a right hemithyroidectomy for thyroid carcinoma. Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) of a tumor in the right lower lobe, adjacent to the inferior pulmonary vein, revealed metastasis of thyroid papillary carcinoma. Immunohistochemical examination of histological cores obtained from the EBUS-TBNA proved extremely helpful in confirming the diagnosis. We performed right lower lobectomy with lymph node sampling, and the final pathological diagnosis was papillary carcinoma without lymph node metastasis.

Transbronchial Biopsy Needle Rinse Solution Used for Comprehensive Biomarker Testing in Patients with Lung Cancer

Introduction: Although genetic information is essential for molecular targeted therapy for personalized medicine, tissue sampling for genetic analysis remains challenging. We investigated the utility of bronchoscopic sampling in non–small-cell lung cancer (NSCLC) patients compared with conventional histological materials for multiple genetic analyses. Methods: Patients with NSCLC proven by onsite cytological evaluation during bronchoscopic survey were eligible for this study. After conventional needle aspiration biopsy by flexible bronchofiberscopy of primary lesions or convex-probe endobronchial ultrasound of lymph nodes, the used needle was rinsed with saline, and the ultra-microsample (uMS) was used for cytological diagnosis and genetic analysis. Gene mutations and fusion genes were examined by high-resolution melting analysis and direct sequencing. The results from the uMS and those from conventional histological samples were compared. Results: A total of 134 lesions (48 primary and 86 metastatic) were analyzed. Adenocarcinoma (n = 80), squamous-cell carcinoma (n = 43), and NSCLC (n = 11) samples were pathologically confirmed in histological cores; however, malignancies were detected in only 45 (34%) of the corresponding uMS. In 62 samples, genetic disorders, including epidermal growth factor receptor (n = 21), K-ras (n = 11), and BRAF mutations (n = 1); anaplastic lymphoma kinase (n = 5), receptor tyrosine kinase (n = 1), and RET fusion genes (n = 1); and silent mutations (n = 22), were identified. In total, 1474 molecular tests were performed, and 1464 tests (99.3%) were identical for both histological samples and uMS. Conclusion: Bronchoscopic uMS (biopsy needle rinsed fluids) are useful for multiple genetic examinations in NSCLC.

Comparison between endobronchial ultrasound-guided transbronchial needle aspiration and 18F-fluorodeoxyglucose positron emission tomography in the diagnosis of postoperative nodal recurrence in patients with lung cancer

OBJECTIVES Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) has a high diagnostic value for preoperative mediastinal staging in patients with lung cancer. In this study, the utility of EBUS-TBNA for the pathological diagnosis of postoperative lymph node recurrence was investigated and compared with that of (18)F-fluorodeoxyglucose positron emission tomography (FDG-PET). METHODS Patients who received both EBUS-TBNA and FDG-PET for the diagnosis of postoperative lymph node recurrence were retrospectively investigated. They underwent routine chest computed tomography (CT) follow-up after thoracotomy, and when hilar or mediastinal lymph nodes showed enlargement on CT compared with the previous chest CT, they were referred for FDG-PET and EBUS-TBNA. We compared the diagnostic performance of these two modalities. In addition, pathological findings of the biopsied sample were evaluated precisely and compared with the results of FDG-PET. Positivity for hypermetabolism on FDG-PET was defined as a standardized uptake value (SUV) greater than 2.5. RESULTS A total of 40 patients were retrospectively reviewed. The sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of EBUS-TBNA were 100% for each parameter, whereas those of FDG-PET were 95.8, 12.5, 62.2, 66.7 and 62.5%, respectively. The SUV of true-positive nodes was significantly higher than that of false-positive nodes (P = 0.001). Twenty-two of 24 patients who were confirmed for recurrence by EBUS-TBNA underwent anticancer treatment. The pathological diagnoses of 14 false-positive cases by FDG-PET were chronic inflammation in 12 and non-specific granuloma in 2. CONCLUSIONS The diagnostic yield of EBUS-TBNA is higher than that of FDG-PET when postoperative lymph node recurrence is suspected.

Evaluation of a learning system for endobronchial ultrasound-guided transbronchial needle aspiration

BACKGROUND Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is an established modality for nodal staging in lung cancer; nevertheless, acquisition on effective fiberscope handling and puncture techniques remains challenging. Here, we present a novel EBUS-TBNA learning system protocol and evaluate the ability of physicians trained using this protocol to perform cytological diagnosis and histological sampling. MATERIAL AND METHODS We designed a 5-step learning system as follows: (1) preparation, (2) probe insertion, (3) sonographic observation, (4) TBNA assistant, and (5) TBNA operator. Each trainee must accomplish the first 4 steps before beginning step 5. In step 5, EBUS-TBNA was performed in tandem by the trainee and supervisor. Diagnostic accuracy and success of histological sampling were recorded for each trial; results of the corresponding supervisor served as a control. RESULTS All 11 trainees entered step 5 after completing steps 1-4 over 5-10 trials. A total of 308 nodes were punctured in step 5. The overall accuracy of cytological diagnosis was 91.2% among trainees, and the histological sampling success rate was 85.4%. The diagnostic accuracy increased from 85.4% to 93.9% (p = 0.027) after 12 needle aspiration experiences. The sizes of nodes associated with success and failure were 13.6 and 11.1mm (p = 0.001), respectively. CONCLUSIONS Our EBUS-TBNA learning system provided a satisfactory educational pathway for trainees and can be used to improve accessibility of EBUS-TBNA.

Changes in diagnostic strategies for hilar-type castleman’s disease: Report of two cases

Castleman’s disease (CD) is rare and difficult to diagnose preoperatively. We report two similar cases of hyaline vascular type CD diagnosed preoperatively using different modalities. In the first case, a biopsy specimen taken during diagnostic thoracoscopy revealed lymphatic structure suggestive of CD. In the second case, endobronchial ultrasound-guided transbronchial needle aspiration enabled sampling of histological cores for histological diagnosis. In both cases, the final diagnosis was established by surgical resection.

The impact of combined pulmonary fibrosis and chronic obstructive pulmonary disease on long-term survival after lung cancer surgery.

PURPOSE The purpose of this study was to determine the impact of pulmonary fibrosis (PF) on postoperative complications and on long-term survival after surgical resection in lung cancer patients with chronic obstructive pulmonary disease (COPD). PATIENTS AND METHODS A retrospective chart review was conducted of 380 patients with COPD who had undergone pulmonary resection for lung cancer at the University Hospital between 1990 and 2005. The definition of COPD was a preoperative forced expiratory volume in 1 second/forced vital capacity (FEV1/FVC) ratio of less than 70%; PF was defined as obvious bilateral fibrous change in the lower lung fields, confirmed by computed tomography. RESULTS PF was present in 41 patients (10.8%) with COPD; the remaining 339 patients (89.2%) did not have PF. The preoperative FVC/FEV1 was significantly lower in the group of patients with PF than in the group without (p < 0.05). Acute lung injury and home oxygen therapy were significantly more common in the PF group; however, the 30-day mortality was similar between the groups. The cumulative survival at 3 and 5 years was 53.6 and 36.9%, respectively, in the PF group and 71.4 and 66.1%, respectively, in the non-PF group (p = 0.0009). Increased age, decreased body mass index, advanced pathologic stage, and the existence of PF were identified as independent risk factors for decreased survival. CONCLUSION PF is a risk factor for decreased survival after surgical treatment in lung cancer patients with COPD.

Pattern of Metastasis Outside Tumor-Bearing Segments in Primary Lung Cancer: Rationale for Segmentectomy

BACKGROUND Patterns of intrapulmonary metastasis, particularly metastasis outside tumor-bearing segments, were investigated in lung cancer patients to address the rationale for segmentectomy. METHODS In a consecutive series of patients who underwent resection of two or more pulmonary segments for primary lung cancer, intrapulmonary spread patterns, such as segmental/intersegmental node metastasis and pulmonary parenchymal metastasis, were pathologically examined. RESULTS Eligible 244 lesions included 167 adenocarcinomas, 66 squamous cell carcinomas, and 11 large cell carcinomas. Pathologic stages included 0 to IA (n=111), IB (n=56), IIA (n=31), IIB (n=20), IIIA (n=23), and IIIB to IV (n=3); and N1 (n=26) and N2 (n=22). Intrapulmonary spread was observed in 24 cases (9.8%). Of these, metastasis outside tumor-bearing segments was only observed in 4 cases (1.6%), and such cancer spread was more frequently seen in cases with extrapulmonary (hilar to mediastinal) nodal metastasis (7.9%) than in cases without extrapulmonary metastasis (0.5%; p=0.01). Metastasis outside tumor-bearing segments was not observed in 64 tumors with pure or mixed ground glass opacity features on computed tomography. Although tumor location (peripheral or central/intermediate) was not related to the incidence of metastasis outside tumor-bearing segments, intrapulmonary spread was observed in only 1 of 52 peripheral small (≤20 mm) tumors. CONCLUSIONS Metastasis outside tumor-bearing segments is rarely observed in cases with tumors (1) without extrapulmonary nodal metastasis and (2) with ground glass opacity or peripheral small (≤20 mm) features.

Innovative technique of transbronchial radiofrequency ablation for intrapulmonary tumors: a preliminary study in a rabbit model.

Background:Radiofrequency ablation (RFA) has emerged as a potential alternative for surgery in clinical oncology. This animal experiment was conducted to evaluate the feasibility, safety, and effectiveness of transbronchial RFA in the treatment of lung tumor. Methods:VX2 lung cancer model was established in Japanese white rabbits by transbronchial injection of tissue clot suspension. After waiting for tumor growth to approximately 10 to 20 mm in diameter, transbronchial RFA was performed on VX2 tumors using the Celon-ProCurve microprobe with a 12 mm active tip, a diameter of 1.3 mm, without cooled-tip electrode under the guidance of biplane x-ray scanning. At first, the power of delivery of RFA was increased in a stepwise manner beginning at 1 W/min up to a maximum of 4 W/min, to seek appropriate power deposition. Next, the extent of ablation under determined power deposition was examined for various time periods. The therapeutic efficacy was evaluated by grossly and pathologically 1 week after transbronchial RFA. Results:All rabbits tolerated the experimental procedures well. Transbronchial RFA at 2 W/min for 20 minute was the most effective setting in this study. Application of more than 2 W/min was not technically feasible using this equipment, leading to destruction of the probe due to high resistance. In transbronchial RFA at 2 W/min condition, the extent of ablation depended on the duration of ablation. Conclusions:This study demonstrates the potential of transbronchial RFA therapy for treatment of lung tumors. Probe improvement and additional study will be required for further progress.