FGFR1-4 RNA-Based Gene Alteration and Expression Analysis in Squamous Non-Small Cell Lung Cancer.

While fibroblast growth factor receptors (FGFRs) are involved in several biological pathways and FGFR inhibitors may be useful in the treatment of squamous non-small cell lung cancer (Sq-NSCLC), FGFR aberrations are not well characterized in Sq-NSCLC. We comprehensively evaluated FGFR expression, fusions, and variants in 40 fresh-frozen primary Sq-NSCLC (stage IA3−IV) samples and tumor-adjacent normal tissues using real-time PCR and next-generation sequencing (NGS). Protein expression of FGFR1−3 and amplification of FGFR1 were also analyzed. FGFR1 and FGFR4 median gene expression was significantly (p < 0.001) decreased in tumors compared with normal tissue. Increased FGFR3 expression enhanced the recurrence risk (hazard ratio 4.72, p = 0.029), while high FGFR4 expression was associated with lymph node metastasis (p = 0.036). Enhanced FGFR1 gene expression was correlated with FGFR1 protein overexpression (r = 0.75, p = 0.0003), but not with FGFR1 amplification. NGS revealed known pathogenic FGFR2,3 variants, an FGFR3::TACC3 fusion, and a novel TACC1::FGFR1 fusion together with FGFR1,2 variants of uncertain significance not previously reported in Sq-NSCLC. These findings expand our knowledge of the Sq-NSCLC molecular background and show that combining different methods increases the rate of FGFR aberrations detection, which may improve patient selection for FGFRi treatment.

[Pneumocystosis in a patient with lymphangioleiomyomatosis]. / Pneumocystozowe zapalenie pluc u chorej na limfangioleiomiomatoze

A 47-year-old, non-smoking woman was admitted to the National Tuberculosis and Lung Diseases Research Institute for diagnosis of progressive exertional dyspnoea and numerous small thin-walled, air-filled cysts equally distributed throughout both lungs revealed in HRCT (high resolution computed tomography) examination. Histological assessment of specimens obtained by open lung biopsy revealed proliferation of immature smooth muscle, showing the expression of the antigen HMB45. On this basis, diagnosis of lymphangioleiomyomatosis was established. The disease caused essential ventilation damage of the lungs (FEV1 1.34 L; 39.71% pred, VC 4.02 L; 94.96% pred, FEV1/ /VC 0.33-4 1.81% pred, DLCO 3.65 mmol/min/Kpa 38.35% pred).During the observation, despite the lack of immunological disorders, the patient developed Pneumocystis jiroveci pneumonia (PCP) that was treated with trimethoprimsulfamethoxazole. Lymphangioleiomyomatosis is a rare disease which results from a defect of TSC genes. The disease is not related to immunological defects or disorders. However, the considerable cystic destruction of the lungs can predispose the patient to opportunistic infections such as the one in the presented case.

Benign pulmonary metastasizing leiomyoma uteri. Case report and review of literature.

Benign metastasizing leiomyoma (BML) is a rare condition in middle-aged women with a history of uterine leiomyomata. It is characterized by the proliferation of, usually multiple, smooth muscle nodules. Approximately 100 cases have been reported in the literature, and the lungs were the most common site of metastases. We report a case of 52-year-old obese woman (BMI 31), hospital worker, smoker, admitted to the hospital with exertional dyspnoea, night sweats, loss of weight, and productive cough. Hysterectomy for a uterine leiomyoma was performed 9 years earlier. In addition, a history of two episodes of superficial vein thrombosis 3 and 2 years before admission was noted. Chest X-ray and subsequently CT chest examinations revealed multiple, non-calcified nodules within the middle and lower parts of both lungs. Specimens obtained by transbronchial biopsy (TBLB) and from open lung biopsy displayed benign muscle cell proliferation compatible with BML. The levels of sex hormones were characteristic for the menopause; therefore, observation was advised. Additionally, Streptococcus pneumoniae was cultured from bronchial washing, and bronchitis was diagnosed. Antibiotics, bronchodilators, and mucolytics were administered, and dyspnoea and cough with expectoration were diminished. Two years later pulmonary lesions have been stable; however, she has put on weight. Subsequently the patient has developed deep vein thrombosis with pulmonary embolism. Anticoagulant treatment was introduced, with some improvement.

[Polymorphism in the N-acetyltransferase 2 gene in patients with lung cancer. Short communication]. / Polimorfizm w genie N-acetylotransferazy 2 u chorych na raka pluca. Doniesienie wstepne.

INTRODUCTION: Individual's risk of developing lung cancer depends not only on exposure to tobacco smoke, but also on the activity of enzymes involved in the activation or deactivation of carcinogens. Arylamine N-acetyltransferase (EC 2.3.1.5) is an enzyme involved in biotransformation of xenobiotics, mainly aromatic and heterocyclic amines and hydrazines. The different acetylation phenotypes within a population are derived from mutations in the NAT 2 gene. These mutations influence the activity (specifically resulting in high or low activity) of the NAT enzyme. Some authors have demonstrated lung cancer predisposing role of slow acetylator phenotype, whereas other reported increased lung cancer risk for fast acetylators or neutral effect of the NAT2 polymorphism. The aim of this preliminary report was to determine the NAT2 gene polymorphism in patients with lung cancer. MATERIAL AND METHODS: 39 patients with inoperable lung cancer (29 - NSCLC and 10 - SCLC), median age 59 years (42- -72) entered the study. Acetylation genotype was determined in the genomic DNA using an allele-specific polymerase chain reaction. We investigated four genetic mutations, C481T, G590A, A803G i G857A, of the gene NAT2. RESULTS: There were 10 different NAT2 genotypes among the 39 patients. Fourteen patients with a NAT2*2 4/4, *4/5, *4/6 and *4/7 were classified as fast acetylators; and 25 patients with a NAT2*5/5, *5/6, *5/7, *6/6, *6/7 or *7/7 genotype were classified as slow acetylators. Among the 10 patients with SCLC - 4 were fast acetylators, and among 29 patients with NSCLC dominated slow acetylation type found in 19 patients (genotypes NAT2 *5/5 and NAT2 *5/6). CONCLUSIONS: Among patients with small cell lung cancer, there was no predominance of genotype of acetylation, whereas among patients with non-small cell lung cancer predominated NAT2*5/5 and NAT2*5/6 genotypes (slow acetylators).

[Simultaneous detection of tumor cells and the positive result of genetic test for Mycobacterium tuberculosis in pleural effusion]. / Jednoczesne wykrycie komórek nowotworowych i materialu genetycznego pratków gruzlicy w plynie oplucnowym.

Pleural effusion is a frequently observed lesion in the course of respiratory diseases such as inflammatory process and cancer metastasis. Its cause may be either tuberculosis (the most common extrapulmonary location is the pleura) and malignant disease of the pleura. Confirmation of tuberculosis is often troublesome. The primary site of cancer may be also difficult to find despite the application of difficult diagnostic methods. Below we present history of 79-year-old female in whom carcinomatous cells and positive result of PCR for Mycobacterium tuberculosis in pleural fluid were discovered simultaneously suggesting the tuberculosis and cancer of unknown primary origin.

Primary mediastinal neoplasms: a report of 1,005 cases from a single institution.

BACKGROUND: This study was an epidemiological analysis of all primary mediastinal neoplasms (PMNs) diagnosed between 2000 and 2016 at the National Tuberculosis and Lung Diseases Research Institute, Poland. METHODS: All patients with any mediastinal abnormality were included in the analysis. The patients' age and gender were obtained from the institutional database. RESULTS: From a cohort of 5,108 patients, 3,691 primary mediastinal lesions were found, including 1,005 (19%) PMNs: lymphomas (533, 53% of PMNs), thymomas (215, 21%), neurogenic tumors (NTs) (100, 10%), germ cell tumors (GCTs) (62, 6%), soft tissue tumors (STTs) (47, 5%) and thymic carcinomas/thymic neuroendocrine tumors (TCs/TNETs) (37 in total, 4%). The most frequent lymphomas were classical Hodgkin lymphomas [256] and primary mediastinal large B-cell lymphomas [163]. Type AB [73] predominated in thymomas and squamous cell carcinomas [9] and carcinoids [10] in TCs/TNETs. NTs encompassed mainly schwannomas [49], ganglioneuromas [21] and neurofibromas [10]. The most frequent STTs were hemangiomas [13] and lymphangiomas [11]. Lymphomas, thymomas and NT were more often in women, TCs/TNETs in men (P<0.001). Lymphomas predominated between the 2nd and 4th decade of life, NTs under the 3rd decade and thymic epithelial tumors between the 6th and 8th decade (P<0.001). There was no correlation between the subtypes of thymomas and the patients' gender (P=0.389) but it was found between histology and patients' age: in patients <30 years of age type B2 and B3 thymomas and >70 years of age AB type and micronodular thymomas with lymphoid stroma (P<0.001) predominated. In the group of GCTs half of them were malignant and these were noted exclusively in men. No correlation between subtypes of NTs or TCs/TNETs and patients' age and gender was found (P>0.05). CONCLUSIONS: PMNs are rare conditions thus awareness of basic epidemiology may be very helpful for final diagnosis.

Pulmonary epithelioid haemangioendothelioma--interferon 2-alpha treatment--case report.

A 62-year-old, obese woman, smoking 10 pack/year was admitted to the National Tuberculosis and Lung Diseases Research Institute to diagnose small, round opacities revealed by routine chest X-ray examination. These lesions had been observed for 5 years. The patient had been treated for psoriasis, hypertension, and insulin-independent diabetes. On admission she was in good condition, complaining of a slight productive cough as well as intermittent osteoarticular pain. Physical examination revealed cutaneous psoriatic lesions, slight edema of the lower limbs, and clubbed fingers. Tuberculin test was positive. Chest Computer Tomography scanning showed partially calcified nodules (up to 1 cm in diameter) located in the middle and base areas of both lungs. No evidence of hilar nor mediastinal lymph node enlargement was seen. Lung specimens displayed intraalveolar and intravascular growth of neoplastic cells. Immunohistochemical expression of Factor VIII, CD31 and CD34 antigens was present. Pulmonary epithelioid haemangioendothelioma was diagnosed. After 6 months of observation, progression of the disease was shown. Interferon alpha treatment was introduced. During the therapy, a slight regression of pulmonary changes was noticed and since then stabilization of the disease was observed.

Pulmonary tuberculosis mimicking lung cancer progression after 10 years of cancer remission.

Differentiation between pulmonary tuberculosis and lung cancer is often challenging for clinicians, especially that both conditions can coexist. This is due to the fact that the clinical and radiological symptoms of both diseases can be similar. Our case report presents a patient who was treated for advanced lung cancer 10 years earlier and currently has been hospitalized again because of a strong clinical and radiological suspicion of the cancer progression, but whose final diagnosis was tuberculosis.

[A comparative evaluation of immunohistochemical markers for the differential diagnosis between malignant mesothelioma, non-small cell carcinoma involving the pleura, and benign reactive mesothelial cell proliferation]. / Ocena przydatnosci reakcji immunohistochemicznych w róznicowaniu miedzybloniaka zlosliwego oplucnej z rakami niedrobnokomórkowymi naciekajacymi oplucna i odczynowym rozrostem miedzyblonka.

INTRODUCTION: Histopathological diagnosis of malignant mesothelioma (MM) and differentiating it from tumors infiltrating the pleura is very difficult. Distinguishing benign reactive mesothelial cell proliferation from MM also presents problems. The objective of this study was to evaluate the significance of selected immunohistochemical stains in differentiating MM from non-small cell lung cancers infiltrating the pleura and from benign reactive mesothelial cell proliferation. MATERIAL AND METHODS: The material encompassed 86 cases of MM, 54 cases of NSCLC infiltrating the pleura, and 43 cases of benign reactive mesothelial cell proliferation. The MM cases were reclassified according to the WHO criteria (2004): epithelioid, 61 cases (71%), including well-differentiated papillomatous, 3 cases; sarcomatous, 6 cases (6.8%); fibrous, 4 cases (4.7%); biphasic, 15 cases (17.5%). A panel of immunohistochemical stains was used in this study. It included broad-spectrum antibodies to cytokeratins (CKAE1/AE3, CKMNF116), vimentin, epithelial membrane antigen (EMA), mesothelial cells (HBME1, CK5/6, calretinin), adenocarcinoma cells (BerEp4, B72.3, CEA, TTF1), antibodies enabling the assessment of proliferation (Mib1) and cell-cycle regulating proteins (p53). RESULTS: Coexpression of cytokeratins and vimentin was found in 63.9% of MM cases and cell-membrane reactions with EMA were seen in 58.9%. Positive staining for HBME1, CK5/6, calretinin, BerEp4, B72.3, CEA and p53 was obtained in 76.7%, 51.2%, 66.7%, 1.2%, 6.2%, 1.2% and 51% of the cases, respectively. None of the MM cases stained for TTF1. MM by WHO subgroups: Coexpression of cytokeratins and vimentin occurred in 55.7% cases of epithelioid MM, 93.3% of biphasic MM, 66.6% of sarcomatous MM, and in 100% of fibrous MM cases. Positive staining for HBME1, CK5/6, and calretinin was seen only in the epithelioid and mixed subtypes of MM; the respective percentages of positive reactions were: HBME1, 90.2% and 73.3%; CK5/6 58.2% and 53.3%; calretinin, 72% and 75%. Non-small cell lung cancers infiltrating the pleura: Coexpression of cytokeratin and vimentin was found in 17.6% of the cases, positive staining of membranes for EMA, in 13% cases. Positive staining for HBME1 was observed in 22.6% of the cases, for CK5/6, in 9.3%, for calretinin, in 2%, for BerEp4, in 72.2%, for B72.3, in 64.1%, for CEA, in 58.5%, and for TTF1, in 43.8%. Benign reactive mesothelial cell proliferation: Protein p53 was present in 9.3% of cases, whereas no positive staining for EMA was found. Differentiation of MM from non-small cell carcinomas: Among the antibodies used in the study, anti-HBME1 had the highest sensitivity (76.7%) but lowest specificity (77.4%). Staining for calretinin showed high specificity (99.8%), as did CEA and TTF1 (98.8% and 100%), with moderate sensitivity (66.7%, 58.5% and 43.8%, respectively). BerEp4 showed the highest sensitivity (72.2%) and specificity (98.8%). CONCLUSION: In diagnosing mesothelioma it is necessary to use a panel of immunohistochemical stains, which should contain antibodies to markers for adenocarcinoma and mesothelioma. Due to the high costs of such a study, a two-stage method is advantageous. The best combination of sensitivity and specificity was found for BerEp4, CEA, and TTF1 and for calretinin and HBME1. In the diagnosis of spindle-cell pleural tumors and the fibrous form of MM and benign reactive mesothelial cell proliferation , markers of mesothelial cells are noncontributory. Immunohistochemical staining fails to identify a reactive process, but a diffuse, positive stain for EMA and the presence of protein p53 support the diagnosis of MM.

Cryptogenic organizing pneumonia-Results of treatment with clarithromycin versus corticosteroids-Observational study.

BACKGROUND: Cryptogenic organizing pneumonia (COP) is a clinicopathological syndrome of unknown origin. Corticosteroids are the standard treatment, but clarithromycin (CAM) is also effective. The aim of this observational retrospective study was to compare the results of CAM versus prednisone (PRE) treatment in patients with biopsy-proven OP without respiratory insufficiency. MATERIAL AND METHODS: In a 15-year period, 40 patients were treated with CAM (500 mg twice daily orally for 3 months) and 22 with PRE (mean initial dose of 0.67 ± 0.24 mg/kg/d for a mean of 8.59 ± 3.05 months). RESULTS: The clinical presentation, laboratory, and radiological findings did not differ markedly between patients treated with CAM and PRE, with the exception of a higher frequency of sweats (55% vs. 23%; p < 0.015), ground glass opacities (95% vs. 50%; p <0.0001) and nodular lesions (45% vs. 18%; p = 0.036) in the CAM group. A complete response was achieved in 35(88%) patients treated with CAM and in all treated with PRE. Patients treated with PRE relapsed more frequently than those treated with CAM (54.5% vs. 10%; p < 0.0001). Corticosteroid-related adverse events were noticed in 8(6.5%) patients (with one death), but CAM caused only one (2.5%) allergic reaction. A FVC >80% identified patients who might be successfully treated with CAM with a sensitivity of 60% and a specificity of 88.57% (AUC 0.869; 95% CI 0.684-1; p = 0.008); the figures for the FEV1 were >70%, a sensitivity of 60%, and a specificity of 91.43% (AUC 0.809; 95%CI 0.609-1; p = 0.027). CONCLUSIONS: CAM can be used to treat COP patients in whom the pulmonary function parameters are within normal limits. Such therapy is shorter, better tolerated, and associated with fewer adverse events and relapses than is PRE. However, the therapy is ineffective in some patients.

D2-40 Antibody is a Specific Marker for B2 Thymomas.

The correlation between the worse outcome of thymomas and expression of podoplanin (D2-40 antibody) in neoplastic cells has been proved in recent studies. We investigated the expression of podoplanin in thymic epithelial tumors of different histologic types and stages resected in our institution. The presence and type of reaction (membranous or cytoplasmic) with D2-40 antibody were assessed. Analyzed group consisted of 72 tumors: 3 type A, 19 type AB, 5 type B1, 24 type B2, 4 type B3, 2 micronodular, 1 metaplastic, and 9 combined B2B3 thymomas and 5 thymic carcinomas. Positive reaction with D2-40 was detected in 7 cases (37%) of AB type, 2 (40%) of B1, 28 (85%) of B2, 3 (23%) of B3, and 1 case (100%) of metaplastic thymoma. All positive B2 and 2 cases of B3 thymomas revealed membranous type of reaction, whereas other subtypes showed less conspicuous cytoplasmic reactivity. A correlation between B2 thymoma and membranous type of reaction was statistically significant (P<0.0001). There was also a slight relationship between cytoplasmic type of reaction and AB or B1 thymomas (P=0.0256). No correlation was detected between D2-40-reactivity and stage (P=0.4) or myasthenic symptoms (P=0.21). In conclusion, membranous type of reaction with D2-40 antibody in thymomas is highly specific for B2 subtype, but antipodoplanin immunoreactivity has no relationship with the tumor stage.

[Granular cell tumor--a rare, benign respiratory tract neoplasm in the material of the Institute of Tuberculosis and Lung Diseases]. / Granular cell tumor--rzadki, lagodny nowotwór ukladu oddechowego w materiale Instytutu Gruzlicy i Chorób Pluc.

The granular cell tumor (GCT) is a nodule that arises most commonly in the skin, the breast or the tongue. The vast majority are benign. Approximately 6-10% of granular cell tumors have been reported in the lower respiratory tract. The clinical, pathological and immunohistochemical findings of eleven cases are described in our material consisted of 6 males and 5 females aged from 35 to 58 years (median, 46 years). The GCT were solitary lesions in all our patients. The tumors were located in trachea (6 cases) and in bronchus (5 cases). They were found during bronchoscopy performed because of symptoms of pneumonia, lung cancer and hemoptysis or dyspnea alone. Diameter of the tumors ranged from 0.2-2.5 cm (median 1.2 cm). Six tumors were surgically excised and 5 were endoscopically removed. Pulmonary GCT behave in a benign fashion. It was observed that tumors of less than 8 mm were more amenable to endoscopic removal and larger tumors were more likely to infiltrate through the bronchial wall. Histologically, the GCT showed submucosal infiltrates of round or oval cells with abundant granular cytoplasm. The tumors cells were positive for S-100 protein, neuron specific enolase, CD68 and vimentin. Our immunohistochemical results are consistent with this concept.

Diagnosis of malignant pericarditis: a single centre experience.

BACKGROUND: Malignancy is the most common cause of effusive pericarditis with a haemodynamically significant amount of pericardial fluid. Early diagnosis and management of malignant pericarditis may significantly improve outcomes. AIM: To evaluate retrospectively the rate and clinical presentation of malignant pericarditis among patients undergoing invasive treatment, with a view to identification of optimal diagnostic modalities to distinguish this group among other patients. METHODS: We studied 191 patients (100 men and 91 women, median age 57 years, range 19-88 years) with effusive pericarditis who underwent invasive treatment in the National Institute of Tuberculosis and Lung Diseases in Warsaw in 1982- -2008 due to a significant amount of pericardial fluid and/or echocardiographic evidence of cardiac tamponade. Pericardiocentesis was performed in 93 cases, pericardioscopy in 61 cases, and substernal pericardiotomy in 37 cases. Pericardial fluid was sent for examination in all patients, and a pericardial specimen was obtained in 96 patients. The patients were divided into 3 groups: Group 1 included patients with malignant pericarditis (malignant cells found in the cytological examination of the pericardial fluid and/or neoplastic infiltration in the histological examination of the pericardial specimen), Group 2 included patients with probable malignant pericarditis (pericardial fluid without malignant cells with histologically confirmed malignancy at some other location), and Group 3 included patients with non-malignant pericarditis (negative cytological examination of pericardial fluid and histological examination of the pericardial specimen, with no evidence of malignancy during hospitalization and one-year follow-up). RESULTS: Malignancy was found in 111 (58%) of 191 patients, including 66 (35%) patients with definite malignant pericarditis and 45 (23%) patients with probable malignant pericarditis. Lung cancer, including adenocarcinoma, was the most common type of malignancy, present in 44 (67%) patients. Non-malignant pericarditis was found in 80 (42%) patients. Among patients with the diagnosis of malignancy (Groups 1 and 2), a positive result of the cytological examination of the pericardial fluid was obtained in 52 cases (sensitivity of 46%). Among patients without malignancy, a negative result of the cytological examination of the pericardial fluid was obtained in all 80 cases (specificity of 100%). Malignant infiltration was found in 20 of 44 patients with the diagnosis of malignancy (sensitivity of 46%) and in none among 52 patients without malignancy (specificity of 100%). Compared to patients with non-malignant pericarditis, patients with malignant pericarditis significantly more commonly presented with tachycardia of >100 bpm in a resting electrocardiogram (ECG) (in 77% of patients with malignant pericarditis vs. 43% of patients with non-malignant pericarditis, p = 0.01), low QRS amplitude (52% vs. 34%, respectively, p = 0.03), electrical alternans (19% vs. 3%, respectively, p = 0.001), echocardiographic evidence of cardiac tamponade (67% vs. 34%, respectively, p = 0.0001), enlarged mediastinal lymph nodes by chest computed tomography (CT) (90% vs. 29%, respectively, p <0.00001), pericardial thickness >8 mm by chest CT (62% vs. 16%, respectively, p <0.0001), and bloody pericardial effusion (94% vs. 43%, respectively, p <0.0001). Levels of carcinoembryonic antigen (CEA) and cytokeratin fragment-19 (CYFRA 21-1) in the pericardial fluid were higher in patients with malignant pericarditis compared to patients with non-malignant pericarditis, with median values of 40.8 ng/mL vs. 0.9 ng/mL, p <0.0001, and 162.85 ng/mL vs. 13.35 ng/mL, p <0.0001, respectively. CONCLUSIONS: 1. Malignancy was found in 58% of patients undergoing invasive treatment due to large pericardial effusion. 2. Cytological examination of the pericardial fluid and histological examination of a pericardial specimen showed high specificity (100%) but low sensitivity (46%) in the diagnosis of malignant pericarditis. 3. The most important predictors of malignant pericarditis included tachycardia of >100 bpm as revealed by the physical examination and ECG, echocardiographic evidence of cardiac tamponade, presence of enlarged mediastinal lymph nodes (>1 cm) and thickened pericardium (>8 mm) by chest CT, bloody pericardial effusion, and elevated levels of CEA (>5 ng/mL) and CYFRA 21-1 (>50 ng/mL) in the pericardial fluid.