Morphological, immunohistochemical, and genetic analyses of bronchiolar adenoma and its putative variants.

We collected 26 cases of bronchiolar adenoma (BA) and its variants, and performed a comprehensive characterization using a combination of morphological, immunohistochemical, and genetic assessments. Of these 26, 13 were classic bilayered cases, including 10 proximal and 3 distal-type BAs. Of note, we also identified 13 cases that lacked a continuous basal cell layer. In five cases, the adenomas were partially classic bilayered, leaving a single layer of columnar or cuboidal epithelial cells in some areas of the lesion (BA with monolayered cell lesions). In the other eight cases, the glandular or papillary structures were entirely composed of monolayered columnar or cuboidal epithelial cells, which were morphologically identical to the luminal epithelial cells of classic BA (monolayered BA-like lesions). Immunohistochemical analysis revealed thyroid transcription factor 1 expression by ciliated columnar epithelial cells, basal cells, and nonciliated columnar and cuboidal epithelial cells. Basal cells also expressed p40 and p63. Twenty-five cases underwent next-generation sequencing using a 422-cancer-gene panel (GeneseeqPrime). Oncogenic driver mutations were detected in 23 cases, including 13 (52%) with EGFR mutations, 4 (16%) with KRAS G12D/V mutations, 3 (12%) with BRAF V600E mutations, 2 (8%) with ERBB2 exon 20 insertions, and 1 (4%) with a RET fusion. EGFR exon 20 insertions were present in 100% of BAs with monolayered cell lesions, 37.5% of monolayered BA-like lesions, and 8% of classic BA (Fisher's exact test, p = 0.002, false discovery rate = 0.014). Collectively, our study revealed a gradual morphological transition between BA and its variants. The genetic composition of BAs with monolayered structures differed significantly from those of classic BAs or lung adenocarcinoma.

[Clinicopathologic features of atypical spindle cell lipomatous tumor].

Objective: To investigate the clinicopathologic characteristics, immunophenotype, differential and diagnostic features of atypical spindle cell lipomatous tumor (ASLT). Methods: Three cases of ASLT were collected from January 2010 to March 2017 at Zhejiang Provincial People's Hospital. The clinical and imaging features, histomorphology, immunophenotype and prognosis were analyzed. Fluorescence in situ hybridization (FISH) was used to detect MDM2 gene amplification, and relevant literature was reviewed. Results: All three patients were adult males, aged 38, 43 and 54 years, respectively. One tumor originated in the subcutaneous soft tissue in the head and neck, one was located in the left primary bronchus and one in the latissimus dorsi muscle. Grossly, all three tumors were circumscribed and ranged from 4.0 to 5.8 cm in size. Microscopically, all showed a focally infiltrative front. These tumors were composed of variable proportions of spindle-shaped and adipocytic cells in a background of variable fibrous and edematous matrix. Scattered lipoblasts were easily seen. One tumor was composed predominately of spindle tumor cells, one of adipocytic cells, and one of equally mixed cell populations. The spindle tumor cells were generally bland-appearing with focal nuclear enlargement and hyperchromasia noted in one case. Mitosis was not seen in neither the spindle cells nor the adipocytic cells. By immunohistochemistry, diffuse and strong reactivity to CD34 of the spindle cells was noted in all cases, definite loss of Rb expression was noted in one of three cases, and S-100 protein was expressed only in the adipocytic cells. INI-1 was intact and Ki-67 index was 1% to 3%. All other markers including CDK4, MDM2, STAT6, SOX10, CD99, bcl-2, ß-catenin, CD117, GFAP, CK, EMA, SMA and desmin were negative. FISH of MDM2 was done in two cases, and both showed no amplification. The ASLT in the head and neck had two recurrences during 17 months of follow-up, whereas the tumor in the latissimus dorsi was free of disease during 33 months of follow-up. Conclusions: ASLT is a rare subtype of low-grade adipocytic neoplasm and is distinctive from atypical lipomatous tumor/well-differentiated liposarcoma. The histomorpholgy of ASLT has significant heterogeneity and forms a continuous spectrum. ASLT needs to be distinguished from a series of benign and malignant soft tissue tumors.

Adenoid cystic carcinoma of the right main bronchus showing squamous differentiation and mimicking mucoepidermoid carcinoma: a case report.

Complete dissection of tracheobronchial adenoid cystic carcinoma (TACC) by surgery alone is sometimes difficult and has a greater propensity than tracheobronchial mucoepidermoid carcinoma (TMEC) for its surgical margin to become positive. In addition, TACC is more likely to present distant metastases than TMEC. Considering these facts, TACC and TMEC should be differentiated based on histopathological examination of biopsy specimens. Herein, we present a case of 54-year-old woman with a tumor in the right main bronchus, whose biopsy specimen was difficult to diagnose as TACC or TMEC. The specimen from the rounded protrusion of the tumor showed squamous differentiation, along with the presence of glandular and basaloid cells, making morphological examination alone ineffective in rendering a definite diagnosis. Thus, the addition of immunohistochemical analysis, αSMA and CD43 expression in basaloid cells and c-kit expression in glandular cells, was useful for accurately diagnosing TACC in this case. The squamous component was considered to be neoplastic because of its increased expression of cyclin D1 and overexpression of p16. The surgically resected specimen contained typical morphology of ACC, and the diagnosis of TACC was definitely confirmed.

Endobronchial metastasis from hepatocellular carcinoma - a case description with literature review.

Endobronchial metastases from hepatocellular carcinoma are very rare. Up to date, no more than 7 cases were reported. The authors present a case of 20-year old female with metastatic hepatocellular carcinoma to superior lobar bronchus. Examination of cytological and small biopsy specimens obtained from bronchoscopy revealed characteristic microscopic features and immunohistochemical profile of hepatocellular carcinoma.

Primary pure signet-ring cell carcinoma of the extrapulmonary left main bronchus: a case report with an immunohistochemical study.

Although several case reports and series of primary signet-ring cell adenocarcinoma (SRCA) of the lung have been reported, primary SRCA of the extrapulmonary main bronchus has not been reported. A 61-year-old man was found to have a tumor of the mediastinum of the left pulmonary hilus on routine chest X-ray examination. A transbronchial endoscopy revealed an elevated tumor in the extrapulmonary left bronchus near the tracheal bifurcation. Biopsy was taken from the bronchial lesion. It revealed pure typical SRCA. Histochemically, Alcian-blue/PAS stain showed intracytoplasmic mucins. Immunohistochemically, the tumor cells were positive for cytokeratin (CK) AE1/3, CK CAM5.2, CK7, CK18, CEA, EMA, CA19-9, Ki-67 (labeling=20%), p53, and MUC1. They were negative for CK34BE12, CK5/6, CK8, CK14, CK19, CK20, p63, vimentin, TTF-1, CDX-2, MUC2, MUC5AC and MUC6. The pathological diagnosis of primary SRCA of the left main bronchus was made. The patient died of carcinomatosis 18 months after the first presentation. In conclusion, the author reported the first case of primary SRCA of the extrapulmonary left main bronchus near the tracheal bifurcation with an extensive immunohistochemical study.

A framework for quantitative assessment of Ki67 distribution in preneoplastic bronchial epithelial lesions.

OBJECTIVE: Deregulated cell proliferation is a hallmark of cancer, and Ki67 immunostaining can be used to identify proliferating cells. Evaluation of cell proliferation may have utility as a biomarker of epithelial malignant transformation risk. To date, most analyses of Ki67 staining have been restricted to semiquantitative estimations of the degree of staining or the measurement of the fraction of Ki67-positive cells within the epithelium. We sought to develop a robust, objective means of quantitatively evaluating Ki67 immunostaining for lung precancerous lesions. STUDY DESIGN: We quantified the spatial distribution of Ki67-expressing cells within the epithelium by means of (1) a cell-based Voronoi tessellation and (2) a basement membrane-referenced distance transform. This was undertaken in a large cohort of 613 lung biopsy sections representing normal, hyperplasia, squamous metaplasia and mild, moderate and severe dysplasia. For each section 21 features quantifying different aspects of the Ki67 staining were calculated. Intraobserver and inter-observer variation were recorded for a subset of the biopsy sections. We examined the behavior of each feature with respect to histopathological grade. RESULTS: These measures demonstrated that proliferation is generally limited to layers 2, 3 and 4 of the epithelium (layer 1 being the basal layer). The proliferation in the basal layer is limited and does not increase with increasing grade of dysplasia. Interobserver and intraobserver effects on these features were assessed, and several were more robust with respect to measuring Ki67 expression pattern than the commonly used fraction of Ki67-positive cells. CONCLUSION: Many of these quantitative features showed associations with histological grade that were as strong as the association that exists based on the fraction of Ki67-positive cells while being much more robust to interobserver- and intraobserver-associated variations. The measured spatial distribution of proliferating cells statistically demonstrated asymmetric cell division behavior in cells in the basal layer, a pattern attributed to stem cells giving rise to transient amplifying cells.

[Somatostatin receptor scintigraphy in pediatric bronchial carcinoid tumor]. / Gammagrafía con análogos de la somatostatina en el diagnóstico y el seguimiento del carcinoide bronquial pediátrico.

INTRODUCTION: Carcinoid tumor is a rare neuroendocrine neoplasm with different locations, the most frequent ones during the pediatric age being the appendix and lung. Scintigraphy with (111)In-DTPA-d-Phe(1)-octreotide has led to an importance advance in the diagnosis of extension in carcinoid tumor patients. We present three pediatric patients with bronchial carcinoid studied with somatostatin analogue scintigraphy (SSRS). CLINICAL CASES: The first patient (9 years) was studied using the SSRS after surgery due to carcinoid tumor in the right lower lobe in which tumor remains was observed (this being clearer in the tomography study). The second patient (10 years) presented due to endobronchial tumor in the left lower lobe together with atelectasis of the LUL and emphysema of the LLL. Radiology imaging techniques suggested the differential diagnosis between the endobronchial carcinoid tumor or plasma cells or foreign body gramuloma. The SSRS showed an abnormal deposit of activity in the left hemithorax consisted with carcinoid tumor. No other areas suggesting metastasis were observed. After the surgery (endobronchial resection), new controls with SSRS showed absence of disease. The third patient (12 years) came after a lobectomy (RUL) due to bronchial carcinoid. The SSRS did not show any abnormal areas of activity. In the subsequent control (3 months), a deposit of activity was observed in the middle third of the right hemithorax, after which a lobectomy was performed (RLL and ML) that showed small remains of neuroendocrine carcinoid. Subsequent controls were negative. CONCLUSION: The SSRS has demonstrated great utility in the diagnosis, follow-up and staging of pediatric patients, carriers of neuroendocrine carcinoid tumors.

Large bronchial granular cell tumor.

We report a 20-year-old woman who underwent complete resection of a granular cell tumor (GCT). On chest computed tomography (CT) scan, a mass with a maximum diameter of 36 mm at the lower bronchus with atelectasis of the right lower lobe was noted. Bronchoscopic examination revealed a whitish mass in the truncus intermedius, and the middle and lower bronchus were unable to be seen. A cytopathological examination of the mass revealed GCT. A right middle and lower lobectomy was performed via a posterolateral thoracotomy. Microscopically, the tumor was composed of polygonal cells with oxyphilic granular cytoplasm and small ovoid nuclei. The cytoplasm of the neoplastic cells was positive for S-100 protein and neuron-specific enolase. The patient's postoperative course was uneventful, and she was asymptomatic after 4 months. A large bronchial GCT is rare, which is why we report this case.

[ACTH-dependent Cushing syndrome, a diagnostic challenge]. / Síndrome de Cushing dependiente de ACTH, un problema diagnóstico.

ACTH dependent Cushing syndrome accounts for approximately 80% of all Cushing syndrome. Distinguishing the pituitary origin from ectopic one is still a diagnosis problem in some cases, especially in bronchial carcinoids, because approximately 50% show cortisol suppresion after dexametasone and may be small and, therefore, difficult to detect in chest radiography. We present a case of a patient with ACTH dependent Cushing syndrome of difficult localization with usual diagnosis tests.

Immunohistochemical markers of proliferation and vascularisation in preneoplastic bronchial lesions and invasive non-small cell lung cancer.

Autofluorescence bronchoscopy (AFB) has been shown to be sensitive to detect preneoplastic lesions in central lung airways system. In early stages of carcinogenesis, up-regulation of cyclooxygenase (COX)-2, Ki67 and/or increased angiogenesis may play a role by promoting the proliferation of tumoral cells and their resistance to apoptosis, as well as angiogenesis, tumor cell invasion and setting up of the metastatic process. The present study compared the expression of proliferative (COX-2, Ki67 and PCNA) and angiogenic markers (CD34 and NG2) between preneoplastic bronchial squamous dysplasia lesions and invasive squamous cell carcinoma. Biopsies obtained during AFB [preneoplastic lesions: low-grade (lesions up to moderate dysplasia), n=13; high-grade lesions (severe dysplasia), n=12] and surgical specimens (resections of bronchogenic carcinoma, n=11) were stained with COX-2, Ki67, PCNA, CD34 and NG2 monoclonal antibodies. Microvessel density (MVD) was analysed based on anti-CD34 immunostaining. Lesions were positive for COX-2 in 12 out of 25 preneoplastic lesions, and in 10 out of 11 invasive carcinomas (p=0.025). In preneoplastic lesions, the mean percentage of Ki67 positive cells was lower compared to invasive carcinomas (37.4+/-5.8 versus 58.6+/-8.4%, p=0.043). In addition, significant differences in MVD were observed between preneoplastic and NSCLC specimen [35.3 (25.9, 61.9) versus 22.1 (20.1, 32.6), p=0.016]. No differences were observed in the mean percentage of PCNA or NG2 positive cells between preneoplastic lesions and invasive carcinomas. Findings of the present study indicate that increases in COX-2 and Ki67 expression may be associated with the development of bronchogenic carcinomas and possibly with acquisition of an invasive phenotype. In contrast, increased CD34 expression in preneoplastic lesions suggests that increased MVD may represent an early marker of lung carcinogenesis.

In vivo time-resolved spectroscopy of the human bronchial early cancer autofluorescence.

Time-resolved measurements of tissue autofluorescence (AF) excited at 405 nm were carried out with an optical-fiber-based spectrometer in the bronchi of 11 patients. The objectives consisted of assessing the lifetime as a new tumor/normal (T/N) tissue contrast parameter and trying to explain the origin of the contrasts observed when using AF-based cancer detection imaging systems. No significant change in the AF lifetimes was found. AF bronchoscopy performed in parallel with an imaging device revealed both intensity and spectral contrasts. Our results suggest that the spectral contrast might be due to an enhanced blood concentration just below the epithelial layers of the lesion. The intensity contrast probably results from the thickening of the epithelium in the lesions. The absence of T/N lifetime contrast indicates that the quenching is not at the origin of the fluorescence intensity and spectral contrasts. These lifetimes (6.9 ns, 2.0 ns, and 0.2 ns) were consistent for all the examined sites. The fact that these lifetimes are the same for different emission domains ranging between 430 and 680 nm indicates that there is probably only one dominant fluorophore involved. The measured lifetimes suggest that this fluorophore is elastin.

Granular cell tumor arising metachronously in the bronchus and esophagus.

We present a case of granular cell tumor (GCT) occurring in the esophagus 7 years after operation for bronchial GCT. A 59-year-old Japanese man complained of epigastralgia, and endoscopic examination of the upper digestive tract disclosed a submucosal tumor in the lower esophagus. Histological examination of the endoscopic mucosal resection of the esophageal tumor showed a proliferation of neoplastic cells with an eosinophilic and granular cytoplasm. The cytoplasm of the neoplastic cells was histochemically positive for PAS stain and immunohistochemically positive for S-100. This tumor did not fulfill any of the diagnostic criteria for malignancy at either the macroscopic or microscopic level. I believe that this is the first case of GCT occurring metachronously in the respiratory and digestive tracts. Clinicians and pathologists should bear in mind that GCT may arise metachronously in the respiratory and digestive tracts.

Tracheobronchial adenoid cystic carcinoma: a clinicopathologic study of 14 cases.

STUDY OBJECTIVES: To investigate the clinicopathologic features of primary tracheobronchial adenoid cystic carcinoma (ACC), and to examine kit protein (CD117) expression, and a possible correlation between the histologic grade and Ki-67 positivity. DESIGN: Retrospective 10-year study (from 1992 to 2001). SETTING: Referral center for interventional pulmonology. PATIENTS: Fourteen patients referred to our institution for laser resection of tracheal or endobronchial tumors. INTERVENTIONS: Twelve patients were treated primarily by laser resection via bronchoscopy with subsequent radiation therapy. The remaining two patients had tracheal resection with postoperative radiation and radiation therapy alone. RESULTS: Fourteen patients (5 men and 9 women) had an average age of 44 years at diagnosis (age range, 29 to 57 years). Five of 14 patients were smokers (average smoking history, 28.5 pack-years), 8 were nonsmokers, and the smoking history was unknown in 1 patient. Major symptoms at presentation included cough (57.1%), dyspnea (71.4%), and hoarseness (14.3%). Two patients (14.3%) presented with respiratory failure. Eleven patients (78.6%) had one or more recurrences, and 4 patients (28.6%) had metastases, primarily to the lungs. The average time to either recurrence or metastasis was 4.6 years after the initial diagnosis. Five patients (35.7%), all of whom had recurrences and/or metastasis during the course of their disease, died of their disease (average time, 8.2 years). Thirteen of 13 cases stained for CD117 were positive in the tumor cell cytoplasm with membranous accentuation. Our cases included 8 grade I tumors, 4 grade II tumors, and 2 grade III tumors, which did not correlate with the degree of Ki-67 positivity. CONCLUSIONS: Tracheobronchial ACCs in our study were more common in women and nonsmokers who presented with nonspecific respiratory symptoms and followed a clinical course generally comparable to that of tumors in the salivary glands. CD117 expression was present in all cases tested, and Ki-67 stain results did not correlate with the tumor grade.

5-Aminolevulinic acid-induced fluorescence in bronchial tumours: dependency on the patterns of tumour invasion.

5-ALA-induced protoporphyrin IX (PPIX) fluorescence kinetics was quantified by fluorescence microscopy in three-dimensional organ co-cultures of human bronchial epithelium, which were infiltrated by four different lung tumour cell lines (EPLC-M31, LCLC-103H, NCI-H125 and NCI-H841). Corresponding fluorescence measurements were performed in monolayer cultures of these tumour cell lines and BEAS-2B cells as a model for normal bronchial epithelium by flow cytometry. Significant differences of fluorescence intensities (FI) between the tumours were detected in organ co-cultures as well as in single cell measurements. Relative FI values in organ co-cultures (FI(EPLC-32M1)>FI(LCLC-H103)>FI(NCI-H125)>FI(NCI-H841)) did not correspond to the measurements in single cells (FI(LCLC-H103)>FI(NCI-H125)>FI(NCI-H841)>FI(EPLC-32M1)). Histology of organ co-cultures revealed different patterns of invasion and tumour cell densities depending on the tumour type. After correction of FI in the co-cultures to tumour cell density the correlation coefficient for fluorescence values between both models increased considerably. Thus, additionally to distinctive features of 5-ALA metabolism, patterns of tumour invasion may be a factor determining 5-ALA-induced fluorescence. Considering these results, a pronounced heterogeneity of 5-ALA-induced fluorescence might be expected in different bronchial tumours in vivo. This could interfere with the diagnostic reliability of 5-ALA-induced fluorescence for early tumour detection.

Endobronchial large cell neuroendocrine carcinoma.

Lung tumors with neuroendocrine morphology include typical carcinoid, atypical carcinoid, large cell neuroendocrine carcinoma, and small cell carcinoma. The World Health Organization emphasizes the importance of mitotic count in differentiating these tumors. We studied the case of a 58-year-old male nonsmoker with recurrent pneumonia and an endobronchial mass, which was removed by right middle lobectomy. The patient was alive with no recurrent disease at 36-month follow-up. Histologically, the tumor showed well developed neuroendocrine morphology but contained up to 20 mitoses per 10 high-power fields and was therefore diagnosed as a large cell neuroendocrine carcinoma. However, several features, including the carcinoid-like morphology and endobronchial location of the tumor, absence of smoking history, and promising clinical course, were more characteristic of an atypical carcinoid than of a large cell neuroendocrine carcinoma. It may be necessary to redefine histologic criteria to allow a higher mitotic rate for classification as an atypical carcinoid.

In-depth analysis of molecular alterations within normal and tumour tissue from an entire bronchial tree.

Using laser capture microdissection (LCM), fluorescent microsatellite analysis and immunohistochemical analysis, we have constructed a detailed topographical molecular map of the entire bronchial tree surrounding a primary bronchial squamous carcinoma in order to establish the relationship between the molecular damage within the airway and that in the tumour itself. Allelic imbalance was analysed using markers on chromosomes 3, 9, 13 and 17. In addition, immunohistochemical analysis for p53 and cyclin D1 expression was performed. Analysis revealed allelic imbalance at several loci at the tumour site but also in 83% of the histologically normal airway specimens of the upper and lower lobes. The fractional allele loss (FAL) value was statistically higher (0.75+/-0.13) in the tumour site than in the distal site of the upper (0.42+/-0.09) and lower lobes (0.31+/-0.08). Immunohistochemical analysis revealed overexpression of p53 and cyclin D1 protein within histologically normal bronchial epithelium, thus confirming previous reports for their early involvement in lung tumour development. This is to date the largest in-depth study of allelic imbalance using LCM in a single individual. The patterns of allele-specific imbalance observed support a clonal or oligoclonal expansion model of outgrowths throughout the lung. The widespread incidence of genetic changes in the whole of lung most likely represents smoking-induced alterations and emphasize the complexity of the field cancerization concept. Our findings point to the need for in-depth studies of the whole bronchial tree tissue surrounding lung carcinomas, in order to identify the genetic changes that differentiate preneoplastic and neoplastic stages in lung carcinogenesis.

Papillary pneumocytoma of the lung simulating a pleomorphic adenoma.

A rare case of papillary pneumocytoma of the lung is reported. The immunohistochemical positivities for EMA, cytokeratin and TTF-l strongly support the hypothesis that the neoplastic cells are originated from type 2 pneumocytes. The tumour also presented areas displaying stroma of non-specific mixoid appearance, simulating a benign tumour of the salivary gland type. We propose that these mixoid areas constitute structures of metaplastic origin.