Association of NLRP1 Coding Polymorphism with Lung Function and Serum IL-1ß Concentration in Patients Diagnosed with Chronic Obstructive Pulmonary Disease (COPD).

Chronic obstructive pulmonary disease (COPD) is a chronic disease characterized by a progressive decline in lung function due to airflow limitation, mainly related to IL-1ß-induced inflammation. We have hypothesized that single nucleotide polymorphisms (SNPs) in NLRP genes, coding for key regulators of IL-1ß, are associated with pathogenesis and clinical phenotypes of COPD. We recruited 704 COPD individuals and 1238 healthy controls for this study. Twenty non-synonymous SNPs in 10 different NLRP genes were genotyped. Genetic associations were estimated using logistic regression, adjusting for age, gender, and smoking history. The impact of genotypes on patients' overall survival was analyzed with the Kaplan-Meier method with the log-rank test. Serum IL-1ß concentration was determined by high sensitivity assay and expression analysis was done by RT-PCR. Decreased lung function, measured by a forced expiratory volume in 1 s (FEV1% predicted), was significantly associated with the minor allele genotypes (AT + TT) of NLRP1 rs12150220 (p = 0.0002). The same rs12150220 genotypes exhibited a higher level of serum IL-1ß compared to the AA genotype (p = 0.027) in COPD patients. NLRP8 rs306481 minor allele genotypes (AG + AA) were more common in the Global Initiative for Chronic Obstructive Lung Disease (GOLD) definition of group A (p = 0.0083). Polymorphisms in NLRP1 (rs12150220; OR = 0.55, p = 0.03) and NLRP4 (rs12462372; OR = 0.36, p = 0.03) were only nominally associated with COPD risk. In conclusion, coding polymorphisms in NLRP1 rs12150220 show an association with COPD disease severity, indicating that the fine-tuning of the NLRP1 inflammasome could be important in maintaining lung tissue integrity and treating the chronic inflammation of airways.

Promoter methylation status of ASC/TMS1/PYCARD is associated with decreased overall survival and TNM status in patients with early stage non-small cell lung cancer (NSCLC).

BACKGROUND: Lung cancer is the leading cause of cancer-related death worldwide, with 5-year overall survival less than 15%. Therefore, it is essential to find biomarkers for early detection and prognosis. Aberrant DNA methylation is a common feature of human cancers and its utility is already recognized in cancer management. The aim of this study was to explore the diagnostic and prognostic value of the promoter methylation status of the ASC/TMS1/PYCARD and MyD88 genes, key adaptor molecules in the activation of the innate immune response and apoptosis pathways. METHODS: A total of 50 non-small cell lung cancer (NSCLC) patients were enrolled in the study. Methylation of bisulphite converted DNA was quantified by pyrosequencing in fresh frozen malignant tissues and adjacent non-malignant tissues. Associations between methylation and lung function, tumor grade and overall survival were evaluated using receiver-operating characteristics (ROC) analysis and statistical tests of hypothesis. RESULTS: Methylation level of tested genes is generally low but significantly decreased in tumor tissues (ASC/TMS1/PYCARD, P<0.0001; MyD88, P<0.0002), which correlates with increased protein expression. Three CpG sites were identified as promising diagnostic marker candidates; CpG11 (-63 position) in ASC/TMS1/PYCARD and CpG1 (-253 position) and 2 (-265 position) in MyD88. The association study showed that the methylation status of the ASC/TMS1 CpG4 site (-34 position) in malignant and non-malignant tissues is associated with the overall survival (P=0.019) and the methylation status of CpG8 site (-92 position) is associated with TNM-stage (P=0.011). CONCLUSIONS: The methylation status of the ASC/TMS1/PYCARD and MyD88 promoters are promising prognostic biomarker candidates. However, presented results should be considered as a preliminary and should be confirmed on the larger number of the samples.

Size, edge, and stage of NSCLC determine the release of CYFRA 21-1 in bloodstream.

BACKGROUND: The computed tomography (CT) is the "golden standard" for the assessment of lung cancer progression due to its ability to clearly display the radiomorphologic characteristics. As lung cancer mortality is very high, more comprehensive approaches may be needed for its earlier diagnosis. The research hypothesis was to investigate the relation between the CT morphologic characteristics (size, stage, and edges) of pulmonary lesion and the extent of release of a soluble fragment of cytokeratin 19 being a part of the cytoskeleton of lung epithelial cells. METHODS: This is a retrospective study including 246 pulmonary lesions being diagnosed and subsequently treated at the University Hospital Centre Zagreb, Croatia. The information about the relevant clinical, radiological, and laboratory facts was collected at the time of diagnosis in 164 NSCLC patients, 52 patients with pulmonary metastases, and 30 benign cysts. CYFRA 21-1 was determined by electrochemiluminescence immunoassay. The nonparametric statistical methods were applied. RESULTS: There was a positive correlation between the size and CYFRA 21-1 in NSCLC unlike metastases or cysts (p = 0.0001). The highest values of CYFRA 21-1 were seen in advanced stages of NSCLC and lesions with spiculated edges. CONCLUSIONS: The level of CYFRA 21-1 positively correlates with the greatest size of NSCLC measured by CT. The differences in CYFRA 21-1 according to TNM classification are significant (p = 0.0001): higher values were observed in advanced stages and with tumors having spiculated, lobulated, and poorly defined edges. The combination of CYFRA 21-1 and CT may help articulate the malignancy of pulmonary lesions.

Differences in initial NSE levels in malignant and benign diseases of the thoracic wall.

BACKGROUND: Neuron-specific enolase (NSE) is widely used to follow-up patients with small cell lung cancer (SCLC). Since the NSE level can be influenced by a broad range of diseases and disorders a large study should be done to assess its level in various lung and non-lung tumors and benign diseases. METHODS: This research included 328 SCLC patients, 717 non-small cell lung cancer (NSCLC), 50 other thoracic cancers such as tumors of the mediastinum and mesothelioma, 35 non-pulmonary cancers like esophagus, breast and stomach cancer, 205 benign diseases, and 37 healthy individuals. The serum level of NSE was measured at initial diagnosis prior to therapy using electrochemiluminescence immunoassay (ECLIA, Roche Diagnostics). RESULTS: The high levels of NSE in SCLC differed significantly from all other groups. The results imply very good sensitivity of NSE in SCLC and good discriminatory power of NSE between SCLC and NSCLC. CONCLUSIONS: The NSE level in SCLC differs significantly from all other tested groups (p < 0.01). The highest values are seen in SCLC extensive disease. ROC curves revealed good discriminatory power of the initial NSE levels separating SCLC from other lung lesions. NSE can be used as a diagnostic tool for the early recognition of the neuroendocrine component of lung tumors and follow-up of SCLC patients.

ICTP in bone metastases of lung cancer.

Bone metastases often appear in advanced stages of lung cancer. They are the result of modulation of bone metabolism by tumor cells that migrated into bone microenvironment and degraded bone organic matrix. Measurement of C-terminal telopeptide of type I collagen (ICTP) in the serum of subjects with lung cancer with and without bone metastases and healthy population is the way to explore bone resorption. In 343 subjects included in this research ICTP level was significantly higher in the bone metastasis than other two groups. The existence of pathologic fracture significantly increased ICTP level. ICTP showed sensitivity of 66.0% in bone metastases at 95.0% specificity in lung cancer stages IA and IB. ICTP is a good diagnostic marker in detection of bone metastasis of lung cancer. Its level can distinguish lung cancer with and without bone involvement and can be used as an addition to standard techniques used in diagnostics of bone metastasis.

CYFRA 21-1 in non-small cell lung cancer--standardisation and application during diagnosis.

There is no ideal tumour marker at present. The clinical application of CYFRA 21-1 is possible once a thorough standardisation process is carried out. Standardisation is achieved by determining the reference range in asymptomatic population, benign and malignant lung diseases, and benign and malignant diseases of other organs. Furthermore, it depends on knowledge of research population characteristics, patient medical histories and individual diagnostic procedure results, the size of research target samples and the clinically defined control groups. The cut-off level of CYFRA 21-1 for non-small cell lung cancer (NSCLC) is 1.72 ng/mL in the Croatian population. It is based on the clinically applicable sensitivity of 78% and specificity of 95% in benign lung diseases. The cut-off value is verified by clinical findings. For clinicians the level of CYFRA 21-1 is an early sign of NSCLC in relation to all the benign lung diseases and all the benign diseases of other organs, of which it was confirmed that they can influence the above level, provided that NSCLC is verified using standard diagnostic methods. The level of CYFRA 21-1 is also influenced by the time of sampling in relation to other diagnostic invasive procedures. The marker is clinically applicable if clinical findings verify it; otherwise, it is useless. This research has involved 343 healthy persons, 474 patients with a benign disease and 4440 patients with a malignant disease, 2453 of whom suffer from NSCLC. The sensitivity of CYFRA 21-1 in NSCLC is 78%, in squamous cell lung cancer (SQC) 84.6%, in adenocarcinomas (AD) 74.3% and in large cell lung cancer (LCC) 75.3%. The level of CYFRA 21-1 differs significantly between healthy persons, benign and malignant diseases (p<10(-3)). There are differences between the three histological types of NSCLC (p<10(-6)) and according to T and N (p<10(-3)). The level of CYFRA 21-1 prompts clinicians to repeat the clinical procedure during diagnosis, and helps to detect the disease earlier and implement treatment in NSCLC. We have achieved high concordance between marker findings and clinical diagnostic.

Determining the cut-off value of pro-gastrin releasing peptide (ProGRP) in lung cancer according to population characteristics.

The standardisation process consisted of determining tumour marker levels in all relevant population groups which are connected with the biology of the marker in normal and tumourous cells. This makes clinical application possible. ProGRP serum levels were measured in 273 healthy subjects, 176 patients with benign diseases and tumours, 200 with small cell lung cancer (SCLC), 294 with non-small cell lung cancer (NSCLC), 21 with carcinoid tumour, 93 with undifferentiated lung cancer, 35 with mixed SCLC-NSCLC, and 189 with other malignancies. ProGRP levels in patients with SCLC and SCLC-NSCLC were significantly higher than in all the other groups (p = 5.4 x 10(-3)). Moreover, in SCLC patients ProGRP levels significantly correlate with the extent of the disease and the patients' smoking habit. The cut-off level of ProGRP for SCLC is 65.89 pg/mL in the Croatian population. It is based on 96.8% specificity in benign diseases which cause problems in differential diagnosis. The sensitivity of ProGRP was 85% at the time of SCLC diagnosis.