EXPRESSION ANALYSIS OF MICRORNA IN PROSTATE CANCER AND IDENTIFICATION OF NOVEL DIAGNOSTIC BIOMARKER

Introduzione: Il tumore alla prostata (PCa) \ue8 una delle neoplasie pi\uf9 comuni tra gli uomini nel mondo occidentale e, globalmente, rappresenta la sesta causa di morte dovuta a cancro. L'approccio diagnostico attuale si basa sulla misurazione dei livelli sierici dell\u2019antigene prostatico PSA (prostate specific antigen), nonostante recenti studi clinici abbiano dimostrato che questo marcatore non riduce significativamente la mortalit\ue0 associata a PCa. In questo scenario abbiamo ipotizzato che i microRNA possano essere nuovi potenziali biomarcatori di PCa. Scopo del lavoro: L'identificazione di microRNA, coinvolti nella progressione neoplastica del tumore prostatico come nuovi biomarcatori diagnostici aggiuntivi al PSA, prognostici e predittivi di aggressivit\ue0 tumorale. La nostra strategia sperimentale ha previsto l\u2019uso di linee cellulari di prostata non tumorigeniche e a diverso grado di malignit\ue0, modelli murini di carcinoma prostatico e casistiche di pazienti affetti da PCa. Materiali e metodi: Linee cellulari commerciali: analisi dell'espressione globale dei miRNA tramite piattaforma low-density array nelle linee di PCa (LNCap, PC3, DU145), normali o iperplastiche (RWPE-1 e BPH-1). Casistiche cliniche: analisi di miRNA selezionati, in un set di pazienti (n=58) nei tessuti di parenchima normale, pre-neoplastico (prostatic intraepithelial neoplasia, PIN) e tumorale. I risultati ottenuti sono stati correlati a parametri clinicopatologici dei pazienti. I potenziali target proteici dei miRNA selezionati sono stati valutati in una casistica pi\uf9 ampia mediante tissue micro-array (TMA). Modello murino transgenico TRAMP: analisi globale di espressione dei miRNA in ghiandole di PIN e tumorali, e nello stroma associato. Linee cellulari primarie: ottenimento di linee di fibroblasti derivate da resezioni chirurgiche di PCa (n=10). Risultati: Dallo screening nelle linee cellulari abbiamo selezionato 23 miRNA poi valutati nei 58 pazienti. Tredici miRNA hanno mostrato differenze di espressione significative (p<0.05). In particolare, nove miRNA (miR-135b,-193a-5p,-205,-224,-22,-34b,-34c-5p,-452,miR-886-3p) sono risultati progressivamente diminuiti nella progressione neoplastica (N-PIN-PCa). Viceversa, i miR-130a, -218, -532, -542-5p, -489 e let-7c hanno mostrato una diminuzione di espressione nel PIN rispetto al tessuto normale. \uc8 noto che i miR-205, miR-218, miR-224 abbiano come bersaglio proteico RUNX2 (Runt-related transcription factor 2), un fattore di trascrizione coinvolto nel tropismo osseo di cellule metastatiche. La valutazione di RUNX2 nei TMA di prostata ha rivelato che la positivit\ue0 nucleare \ue8 specifica delle cellule tumorali (p<0.0001) e correla con l\u2019estensione del tumore e con l'invasione capsulare. Inoltre il confronto tra l\u2019espressione dei miRNA e RUNX2 ha mostrato correlazione inversa significativa. Dall\u2019analisi del modello TRAMP abbiamo identificato un pannello di miRNA differenzialmente espressi tra le componenti epiteliali e stromali associate a PIN o PCa (n=52). Conclusioni: I nostri risultati mostrano una sinergica perdita di miRNA con funzione oncosoppressiva e contemporaneo aumento di RUNX2 nei tessuti di PCa. Questo dato ha importanti implicazioni a livello di progressione di malattia che sar\ue0 valutata in una successiva fase del progetto. Le nostre analisi hanno indicato il miR-205 come potenziale marcatore di aggressivit\ue0 di malattia. Hanno inoltre identificato 9 miRNA precocemente persi nelle lesioni precancerose (PIN) rispetto al parenchima sano. Studi in casistiche indipendenti potranno confermare queste molecole come nuovi biomarcatori di neoplasia da affiancare alla valutazione del PSA. Inoltre lo studio dei miRNA \u201cstromali\u201d ha evidenziato una profonda deregolazione di queste molecole nel microambiente tumorale rispetto a quello non neoplastico. Questo risultato sottolinea dal punto di vista molecolare l\u2019importanza dello stroma nel sostenere la sopravvivenza e la crescita tumorale e fornisce una possibile strategia terapeutica alternativa, mirata alle cellule stromali anzich\ue9 epiteliali per indurre regressione di malattia. Saranno necessari ulteriori studi per valutare il ruolo dei miRNA nell\u2019interazione tra epitelio tumorale e stroma circostante.Introduction: Prostate cancer (PCa) is one of the most common cancers among men and the sixth cause of cancer-related death in men worldwide. The current diagnostic approach is based on serum measurement of prostate specific antigen (PSA) levels, despite recent clinical studies showed that did not considerably reduced mortality incidence in prostate cancer patients. In this scenario, we hypothesized that microRNAs (miRNAs) could be novel biomarkers for PCa disease. Aim of the study: We propose to identify miRNA signatures associated to PCa progression that could represent a novel generation of diagnostic biomarkers adjunctive to PSA, prognostic and predictive of cancer progression. Our experimental strategy included the use of normal or tumorigenic prostate cell lines, mouse model of PCa and patients\u2019 series. Methods: Prostate cell lines: global miRNA expression analysis using a low-density array platform in PCa (LNCap, PC3, DU145) or non-tumorigenic cells (RWPE-1, BPH-1). Clinical series. Analysis of selected miRNAs in 58 PCa patients for which normal parenchyma and prostatic intraepithelial neoplasia (PIN) was available. Correlation of molecular profiles to clinicopathological characteristics. Potential miRNA targets were investigated using a larger series of PCa patients arranged in tissue micro-arrays (TMAs). TRAMP mouse: global miRNA profiles were obtained from epithelial and stromal compartments of PIN or tumoral lesions. Primary cell lines: fibroblasts were obtained from prostate resection of PCa patients (n=10). Results: miRNA screening in cell lines provided a panel of 23 miRNAs that were then investigated in the 58 PCa patients. Thirteen miRNA displayed significant deregulation (p<0.05) in disease tissues. Specifically nine miRNAs (miR-135b,-193a-5p,-205,-224,-22,-34b,-34c-5p,-452, miR-886-3p) were progressively down-regulated during neoplastic progression (N-PIN-PCa). Conversely, miR-130a, -218, -532, -542-5p, -489 and let-7c displayed lower levels in PIN compared to normal prostate. A recognized target of miR-205, miR-218 and miR-224 is the Runt-related transcription factor 2 (RUNX2), a protein involved in metastatic dissemination to the bone. In our patients\u2019 TMA, RUNX2 was overexpressed in tumoral cell nuclei (p<0.0001) and it was related to tumor size and capsular invasion. Moreover RUNX2 was inversely related to miRNA levels. TRAMP mice analysis has provided a signature of miRNAs (n=52) differentially expressed in epithelial and stromal compartments of PIN or PCa cells. Conclusions: Our results show a simultaneous loss of oncosuppressive miRNAs and increased RUNX2 expression in PCa tissues. This data is particularly relevant in disease progression monitoring, an aspect that will be studied in future project\u2019s phases. Our analysis showed that miR-205 loss is a potential biomarker of aggressive disease. Furthermore, we identified nine miRNAs which expression is decreased from early stage of disease (PIN). Validation of this result in independent patients\u2019 series could provide novel biomarkers of PCa useful as adjunts to PSA monitoring. Lastly, profound stromal miRNAs deregulation underlines the importance of tumour microenvironment in sustaining cancer cell survival and growth. Moreover this result suggest that targeting tumour stroma could represent an alternative strategy for anti-cancer therapies. Future studies are needed to shed light on this aspect

Design and manufacturing of a Nd-doped phosphate glass-based jewel

This paper reports the results of the designing, manufacturing and characterization of a jewel obtained by means of coupling the dogmas of industrial design to the analytical engineering approach. The key role in the design of the jewel was played by an in-house synthesized Neodymium (Nd)-doped phosphate glass, selected due to its easy handling and capability to change color according to the incident light wavelength. The glass core was covered by a metal alloy to mitigate its relatively high fragility and sensitivity to thermal shock and, at the same time, to highlight and preserve its beauty. The selection of the proper metal alloy, having thermo-mechanical properties compatible with those exhibited by the glass, was carried out by means of Ashby's maps, a powerful tool commonly adopted in the field of industrial design

Adherence to recommendations for cervical and breast cancer screening in systemic sclerosis.

The aim of the study was to evaluate the adherence of systemic sclerosis (SSc) female patients to cervix and breast cancer screening procedures, as suggested by local guidelines. A cohort of 84 SSc women was asked if they had undergone mammography and Pap test during the previous 2- and 3-year intervals, as indicated according to the Italian recommendations. The results were compared with those collected in patients affected by other chronic rheumatic disorders and in the general population. More than 85% of SSc women declared to comply with an age-related cervix and breast cancer screening schedule. The data were similar to those collected in patients affected by other chronic rheumatic disorders, whereas the subjects belonging to the general population reported to undergo breast cancer screening more frequently. Among SSc women, neither the educational level, nor the lung and skin involvement influenced their cancer screening program compliance. Only a positive history of ischemic digital ulcers seemed to interfere with mammography. Our study reported a very high percentage of SSc female patients who adhered to programs for the early detection of cervical and breast cancer. The significant adherence to guidelines may be due to the schedule adopted by the local health public service, which regularly invites eligible subjects by mail to undergo cancer screening at no charge

Civil Society Activism in Italy Across Different Fields: A Multifaceted Picture of Solidarity in Hard Times

Over the last years, Italian civil society organisations have been working on a daily basis to mitigate the impact of both the global economic crisis and the refugee crisis, which have increased social vulnerabilities. Relying on the data gathered through 30 in-depth interviews with transnational solidarity organisations’ representatives, this chapter analyses solidarity practices in three fields of activity: disability, unemployment and migration. Results show that solidarity attitudes, practices and discourses are strongly influenced by the policy domain in which the organisations are active. Furthermore, the crisis led organisations to search for new strategies and approaches, even though it has been an ineffective vector of transnationalisation due to lack of resources, and the necessity to cope with pressing needs at national and local level/s.Results presented in book have been obtained through the project ‘European paths to transnational solidarity at times of crisis: Conditions, forms, role-models and policy responses’ (TransSOL). This project was funded by the European Commission under the Horizon 2020 research and innovation programme (grant agreement No. 649435)

po 450 interplay between coding and non coding genome in human parathyroid tumours

Introduction Parathyroid tumours are the second most common endocrine neoplasia in women, after thyroid cancer. Mutations in the oncosuppressor CDC73 are the key event in most carcinomas whereas alterations in the tumour suppressor MEN1 (located at 11q13.1) occur in up to a third of sporadic adenomas. Although lncRNAs play a regulatory role in endocrine cancer pathogenesis, a lncRNAs profiling in human parathyroid tumours is still missing. Here, we identified a 'molecular signature' able to distinguish among parathyroid histotypes and a new potential epigenetic role of MEN1 in lncRNAs regulation. Material and methods Ninety lncRNAs were investigated in 4 parathyroid carcinomas (PCas), 12 adenomas (PAds) and 2 normal glands (PaNs). Hierarchical clustering (HCL) and Significance Analysis of Microarray (SAM) were performed to identify differences in lncRNAs expression. Significant lncRNAs were validated in additional 7 PCas, 26 PAds, 6 atypical PAds (aPAds) and 4 PaNs. CDC73 and MEN1 genes mutations were detected by Sanger sequencing. PAds genomic characterisation was obtained by array Comparative Genomic Hybridization (aCGH). HEK293 cells were transiently silenced for MEN1 expression to analyse MEN1-lncRNAs correlation. Results and discussions Nine lncRNAs were identified as differentially expressed in parathyroid tissues. Specifically, KCNQ1OT1 and SNHG6 were enriched in PaNs, reduced HAR1B, MEG3, HOXA3as and NEAT1 expression characterised PAds, whereas BC200, HOXA6as and WT1-AS were upregulated in PCas. HCL identified 3 clusters in which PaNs and PCas were distinctly separated, while aPAds were closer to PCas. Moreover, PAds clustered in a highly heterogeneous way. Notably, PCas and aPAds harbouring CDC73-mutations overexpressed the majority of the lncRNAs, compared to CDC73 wild-type samples. Interestingly, BACE1-AS, KCNQ1OT1, NEAT1 and SNHG6 levels in PAds were positively correlated with MEN1 levels. aCGH analysis revealed that Chr11 loss of heterozygosity (LOH) was the main chromosomal aberration in PAds. Of note, Chr11 LOH was associated with significant HAR1B upregulation and these data were confirmed in HEK293 cells knocked-down for MEN1. Conclusion Parathyroid histotypes are characterised by different lncRNAs signatures, suggesting a correlation with tumour aggressiveness and pathogenetic mechanisms. Further, our data highlight that lncRNAs profiles are related to CDC73 gene mutation status, chromosome 11 derangements and MEN1 inactivation

Towards a map of the Upper Pleistocene loess of the Po Plain Loess Basin (Northern Italy)

Upper Pleistocene (MIS 4-2) loess sequences occur in most of continental Europe and in Northern Italy along the Po Plain Loess Basin. Loess is distributed along the flanks of the Po Plain and was deposited on glacial deposits, fluvial terraces, uplifted isolated hills, karst plateaus, slopes and basins of secondary valleys. Loess bodies are generally tiny and affected by pedogenesis, being locally slightly reworked by slope processes and bioturbation. Notwithstanding, loess in the Po Plain is an important archive of paleoenviron-mental record and its mapping provides new insights in paleoenvironmental and palaeoseismic reconstructions of Northern Italy

Impact of endometriosis on obstetric outcome after natural conception: a multicenter Italian study

Purpose To evaluate obstetric outcome in women with endometriosis who conceive naturally and receive standard obstetric care in Italy. Methods Cases were consecutive women with endometriosis managed in eleven Italian referral centers. Controls were women in whom endometriosis was excluded. All women filled in a questionnaire addressing previous natural pregnancies. Marginal logistic regression models were fitted to evaluate the impact of endometriosis on obstetric outcome. A post hoc analysis was performed within the endometriosis group comparing women with severe adenomyosis versus women with absent or mild adenomyosis. Results Three hundred and fifty-five pregnancies in endometriosis group and 741 pregnancies in control group were included. Women with endometriosis had a higher risk of preterm delivery &lt; 34 weeks (6.4% vs 2.8%, OR 2.42, 95% CI 1.22–4.82), preterm delivery &lt; 37 weeks (17.8% vs 9.7%, OR 1.98, 95% CI 1.23–3.19), and neonatal admission to Intensive Care Unit (14.1% vs 7.0%, OR 2.04, 95% CI 1.23–3.36). At post hoc analysis, women with endometriosis and severe adenomyosis had an increased risk of placenta previa (23.1% vs 1.8%, OR 16.68, 95% CI 3.49–79.71), cesarean delivery (84.6% vs 38.9%, OR 8.03, 95% CI 1.69–38.25) and preterm delivery &lt; 34 weeks (23.1% vs 5.7%, OR 5.52, 95% CI 1.38–22.09). Conclusion Women with endometriosis who conceive naturally have increased risk of preterm delivery and neonatal admission to intensive care unit. When severe adenomyosis is coexistent with endometriosis, women may be at increased risk of placenta previa and cesarean delivery. Trial registration Clinical trial registration number: NCT03354793

Efficacy and safety of rituximab with and without methotrexate in the treatment of rheumatoid arthritis patients: Results from the GISEA register.

Rituximab (RTX) is a monoclonal anti-CD20 antibody approved for the treatment of rheumatoid arthritis (RA) in association with methotrexate (MTX)