Donatella Ugolini

miR-1254 and miR-574-5p: Serum-Based microRNA Biomarkers for Early-Stage Non-small Cell Lung Cancer

Introduction: The ability to diagnose non-small cell lung cancer (NSCLC) at an early stage may lead to improved survival. The aim of this study was to identify differentially expressed serum-based microRNAs (miRNAs) between patients with early-stage NSCLC and controls. These miRNAs may serve as biomarkers for NSCLC early detection. Methods: miRNA profiling was performed on total RNA extracted from serum obtained from 22 individuals (11 controls and 11 patients with early-stage NSCLC). Quantitative polymerase chain reaction (qPCR) was used to validate the profiling results in the discovery set and in a validation set of 31 controls and 22 patients with early-stage NSCLC. Additionally, six matched plasma samples (four NSCLC cases and two controls) and three serum mesothelioma samples were analyzed by qPCR. Receiver operating characteristic curves were generated for each possible combination of the miRNAs measured by qPCR. Results: The expression of hsa-miR-1254 and hsa-miR-574-5p was significantly increased in the early-stage NSCLC samples with respect to the controls. Receiver operating characteristic curves plotting these two miRNAs were able to discriminate early-stage NSCLC samples from controls with 82% and 77% of sensitivity and specificity, respectively, in the discovery cohort and with 73% and 71% of sensitivity and specificity, respectively, in the validation cohort. The mesothelioma and plasma samples did not seem to classify into either NSCLC or control groups. Conclusions: Serum miRNAs are differentially expressed between patients with early-stage NSCLC and controls. The utility of miR-1254 and miR-574-5p serum-based biomarkers as minimally invasive screening and triage tools for subsequent diagnostic evaluation warrants additional validation.

Drug metabolism polymorphisms as modulators of cancer susceptibility

Recently, several molecular genetic bases of polymorphic enzyme activities involved in drug activation and detoxification have been elucidated. Many molecular epidemiology studies based on these premises have sought to gather information on the association of genetically determined metabolic variants with different risks of environmentally induced cancer. While rare alterations of tumor suppressor genes dramatically raise cancer risk for the single affected subjects, far more common and less dramatic differences in genes encoding for drug metabolism enzymes can be responsible for a relatively small, but rather frequent increase of cancer risk at the population level. This increase could be especially important in specific cases of occupational, pharmacological or environmental exposure. Examination of the current literature reveals that the most extensively investigated metabolic polymorphisms are those of P450 1A1 and P450 2D6 cytochromes, glutathione S-transferases (GSTs; M1 and, to a lesser extent, M3, P1 and T1) and N-acetyltransferases (NATs; NAT1 and NAT2). Making reference to these enzymes, we have assayed the current knowledge on the relations among polymorphisms of human xenobiotic-metabolizing enzymes and cancer susceptibilities. We have found intriguing models of susceptibility toward different types of cancer. We have reviewed and commented these models on light of the complex balance among different enzyme activities that, in each individual, determines the degree of each cancer susceptibility. Moreover, we have found techniques of molecular genetic analysis, more suitable than previous ones on phenotypic expression, now allowing better means to detect individuals at risk of cancer. According to the models presently available, a systematic screening of individuals at risk seems to make sense only in situations of well defined carcinogenic exposures and when performed by the polymorphism analysis of coordinated enzyme activities concurring to the metabolism of the carcinogen(s) in question. Genetic polymorphism analysis can allow for the detection of patients more prone to some types of specific cancers, or to the adverse effects of specific pharmaceutical agents. Considering the increasingly confirmed double-edged sword nature of metabolism polymorphism (both wild-type and variant alleles can predispose to cancer, albeit in different situations of exposure), individual susceptibility to cancer should be monitored as a function of the nature, and mechanism of action, of the carcinogen(s) to which the individual under study is known to be exposed, and with reference to the main target organ of the considered type of exposure.

Ovarian suppression using luteinizing hormone-releasing hormone agonists during chemotherapy to preserve ovarian function and fertility of breast cancer patients: a meta-analysis of randomized studies

BACKGROUND The role of temporary ovarian suppression with luteinizing hormone-releasing hormone agonists (LHRHa) in the prevention of chemotherapy-induced premature ovarian failure (POF) is still controversial. Our meta-analysis of randomized, controlled trials (RCTs) investigates whether the use of LHRHa during chemotherapy in premenopausal breast cancer patients reduces treatment-related POF rate, increases pregnancy rate, and impacts disease-free survival (DFS). METHODS A literature search using PubMed, Embase, and the Cochrane Library, and the proceedings of major conferences, was conducted up to 30 April 2015. Odds ratios (ORs) and 95% confidence intervals (CIs) for POF (i.e. POF by study definition, and POF defined as amenorrhea 1 year after chemotherapy completion) and for patients with pregnancy, as well hazard ratios (HRs) and 95% CI for DFS, were calculated for each trial. Pooled analysis was carried out using the fixed- and random-effects models. RESULTS A total of 12 RCTs were eligible including 1231 breast cancer patients. The use of LHRHa was associated with a significant reduced risk of POF (OR 0.36, 95% CI 0.23-0.57; P < 0.001), yet with significant heterogeneity (I(2) = 47.1%, Pheterogeneity = 0.026). In eight studies reporting amenorrhea rates 1 year after chemotherapy completion, the addition of LHRHa reduced the risk of POF (OR 0.55, 95% CI 0.41-0.73, P < 0.001) without heterogeneity (I(2) = 0.0%, Pheterogeneity = 0.936). In five studies reporting pregnancies, more patients treated with LHRHa achieved pregnancy (33 versus 19 women; OR 1.83, 95% CI 1.02-3.28, P = 0.041; I(2) = 0.0%, Pheterogeneity = 0.629). In three studies reporting DFS, no difference was observed (HR 1.00, 95% CI 0.49-2.04, P = 0.939; I(2) = 68.0%, Pheterogeneity = 0.044). CONCLUSION Temporary ovarian suppression with LHRHa in young breast cancer patients is associated with a reduced risk of chemotherapy-induced POF and seems to increase the pregnancy rate, without an apparent negative consequence on prognosis.

Impact assessment of oncology research in the European Union

In this study the distribution of papers published by authors from the European Union (EU) in oncological journals was analysed, as was the impact of oncological research in the EU compared with that produced in other countries. Papers published during 1995 in the oncological journals listed by ISI (Institute for Scientific Information, Philadelphia, U.S.A.) were downloaded. The parameters of impact factor (IF), source country population and gross domestic product (GDP) were considered. An analysis of the key words, both those reported by the authors and those attributed by ISI, was carried out using a special purpose program. 36.5% of papers published in oncological journals come from the EU (the U.K., Italy, Germany and France ranking at the top) and 40.7% from the U.S.A. The mean IF was 2.4 for EU papers, 3.3 for the US and 2.4 for other countries. Our data confirm that smaller countries performed better than larger ones. The key words analysis shows that the leading fields of research were breast cancer for diseases, cisplatin for drugs and p53 for experimental studies. A standardisation of key words on behalf of journal editors is proposed.

Downregulated MicroRNAs in the Differential Diagnosis of Malignant Pleural Mesothelioma

Malignant pleural mesothelioma (MPM) is a rapidly fatal disease whose diagnosis, particularly through less invasive techniques such as analysis of pleural effusion, can be challenging. Currently, a commercially available diagnostic test based on microRNA (miRNA) expression patterns is purported to distinguish between mesothelioma and lung adenocarcinoma. Yet, the biological basis of this technology has not been reported in the literature, and little research has been aimed at determining how differential miRNA expression contributes to the differences in pathogenesis between these diseases, both of which can be caused by asbestos exposure. We sought to illuminate the molecular differences between mesothelioma and lung adenocarcinoma by using miRNA microarrays to identify patterns in the most differentially expressed miRNAs. From this, we identified a panel of miRNAs, including members of the miR‐200 gene family, that were all downregulated in MPM compared to lung adenocarcinoma. Using the more sensitive detection method of quantitative RT‐PCR on an independent series of tumors, we validated the specificity of these alterations in 100 MPMs and 32 lung adenocarcinomas. Statistical analysis reveals that these miRNAs exceed the current recommendations for biomarkers and could greatly aid in the differential diagnosis. Further examination led us to predict that they act as redundant regulators of wnt signaling and suggests a role for this pathway in tumor progression. This research points to novel approaches using miRNAs whose decreased expression is unique to mesothelioma as potentially suitable for rapid diagnosis and reveals prospective new targets for the treatment of this deadly disease.

Increased risk of lung cancer in individuals with a family history of the disease: A pooled analysis from the International Lung Cancer Consortium

BACKGROUND AND METHODS Familial aggregation of lung cancer exists after accounting for cigarette smoking. However, the extent to which family history affects risk by smoking status, histology, relative type and ethnicity is not well described. This pooled analysis included 24 case-control studies in the International Lung Cancer Consortium. Each study collected age of onset/interview, gender, race/ethnicity, cigarette smoking, histology and first-degree family history of lung cancer. Data from 24,380 lung cancer cases and 23,305 healthy controls were analysed. Unconditional logistic regression models and generalised estimating equations were used to estimate odds ratios and 95% confidence intervals. RESULTS Individuals with a first-degree relative with lung cancer had a 1.51-fold increase in the risk of lung cancer, after adjustment for smoking and other potential confounders (95% CI: 1.39, 1.63). The association was strongest for those with a family history in a sibling, after adjustment (odds ratios (OR) = 1.82, 95% CI: 1.62, 2.05). No modifying effect by histologic type was found. Never smokers showed a lower association with positive familial history of lung cancer (OR = 1.25, 95% CI: 1.03, 1.52), slightly stronger for those with an affected sibling (OR = 1.44, 95% CI: 1.07, 1.93), after adjustment. CONCLUSIONS The occurrence of lung cancer among never smokers and similar magnitudes of the effect of family history on lung cancer risk across histological types suggests familial aggregation of lung cancer is independent of those risks associated with cigarette smoking. While the role of genetic variation in the aetiology of lung cancer remains to be fully characterised, family history assessment is immediately available and those with a positive history represent a higher risk group.

Genetic susceptibility to malignant pleural mesothelioma and other asbestos-associated diseases

Exposure to asbestos fibers is a major risk factor for malignant pleural mesothelioma (MPM), lung cancer, and other non-neoplastic conditions, such as asbestosis and pleural plaques. However, in the last decade many studies have shown that polymorphism in the genes involved in xenobiotic and oxidative metabolism or in DNA repair processes may play an important role in the etiology and pathogenesis of these diseases. To evaluate the association between diseases linked to asbestos and genetic variability we performed a review of studies on this topic included in the PubMed database. One hundred fifty-nine citations were retrieved; 24 of them met the inclusion criteria and were evaluated in the review. The most commonly studied GSTM1 polymorphism showed for all asbestos-linked diseases an increased risk in association with the null genotype, possibly linked to its role in the conjugation of reactive oxygen species. Studies focused on GSTT1 null and SOD2 Ala16Val polymorphisms gave conflicting results, while promising results came from studies on alpha1-antitrypsin in asbestosis and MPO in lung cancer. Among genetic polymorphisms associated to the risk of MPM, the GSTM1 null genotype and two variant alleles of XRCC1 and XRCC3 showed increased risks in a subset of studies. Results for the NAT2 acetylator status, SOD2 polymorphism and EPHX activity were conflicting. Major limitations in the study design, including the small size of study groups, affected the reliability of these studies. Technical improvements such as the use of high-throughput techniques will help to identify molecular pathways regulated by candidate genes.

Occupational exposure to formaldehyde and biological monitoring of Research Institute workers

AIM The aim of this study was to verify the presence of a relationship between formaldehyde exposure in the work environment with biological markers of exposure and of effect. METHODS Exposure to formaldehyde (FA) of 36 workers in different laboratories of a Cancer Research Institute and biomarkers of exposure, such as formaldehyde human serum albumin conjugate (FA-HSA) and biomarkers of effect, such as chromosome aberration (CA), micronuclei (MN) and sister chromatid exchanges (SCEs) were measured in peripheral blood lymphocytes of the same workers. RESULTS Individual FA levels of exposure ranged from 4.9 microg/m(3) to 268.7 microg/m(3). Subjects with high FA exposure showed a significant increase of the biomarker of exposure FA-HSA, but biomarkers of effect did not show any significant differences. CONCLUSIONS A significant relationship was observed between occupational exposure to FA and a biological marker of exposure (FA-HSA). The markers of effect used (CA, MN and SCE) failed to indicate the presence of genetic damage.

Trends in otolaryngology research during the period 1995-2000: a bibliometric approach.

OBJECTIVES: To evaluate the distribution and scope of papers published in the world in otolaryngology (ORL) journals and to compare the impact of this research among different countries. METHODS: Papers published in the 29 ORL journals screened by the Institute for Scientific Information (ISI, Philadelphia, PA, USA) in the 6-year period 1995–2000 were considered. The journal impact factor (IF), the source country population, and gross domestic product (GDP) were recorded. All key words, both those assigned by the authors and those attributed by ISI, were identified and their frequency was calculated using a special-purpose program. RESULTS: The total number of papers in the ORL literature during the period 1995–2000 increased from 2036 to 3705. A percentage varying between 47.7% (1995) and 36.1% (2000) was published by EU authors whereas the USA accounted for a percentage varying between 28.1% (1995) and 38.8% (2000). In 2000, the leading countries were the USA, the EU, Japan, Canada, and Australia. In Europe the UK (28.5% of papers), Germany (26.2%), Italy (7.2%), Sweden (5.8%), France (5.5%), and the Netherlands (4.9%) showed a very good performance trend. In the same year, the mean IF of EU papers was 0.8 in comparison with 1.1 for Australia and the USA and 0.9 for the world. In 1997, 1341 key words attributed by the authors and 696 attributed by ISI appeared in the ORL literature. Less than a tenth of them were cited more than twice. The leading key words were “cancer” for disease and “surgery” for treatment. CONCLUSIONS: Bibliometric findings are useful to follow research trends. Our data show high scientific production of relatively small countries. Dispersion of key words should be avoided and journal editors should promote their standardization.

How the European Union writes about ophthalmology

This study evaluates the distribution of papers published by European Union (EU) authors in ophthalmological journals from 1995 to 1997. The impact of ophthalmological research in the EU is compared with that produced in other countries and trends of research are highlighted through the keywords analysis. Data of articles published in ophthalmological journals (ISI Subject Category) were downloaded. Mean Impact Factor, source country population and gross domestic product were analyzed. A special purpose software for keyword elaboration was utilized. 11,219 papers were published in the world in the ophthalmological journals: 34.8% came from the EU (UK, Germany, France, Italy and the Netherlands ranking at the top) and 40.7% from the US. The mean Impact Factor of EU papers was 0.8 in comparison with 1.5 in the US. Despite the limitations of the existing methods, bibliometric findings are useful for the monitoring of research trends. The keywords analysis shows that the leading fields of research were retinal pathologies for diseases and keratoplasty for surgical procedures. It also suggests that keywords are overused, and urges minimization of this as well as standardization among journal editors.