Validation of novel biomarkers for colorectal cancer detection and production of novel antibodies against E-selectin ligands

In 2018, colorectal cancer (CRC) remains the second deadliest kind of cancer with 881,00 deaths of the 1.8 million new cases. Late stages detection is more likely to develop recurrences, even after treatment, leading to the necessity to create a new system to early stages detection. Thus, understanding the biology of the cancer and biomarker discovery are important steps in cancer research. Several studies on cancer-associated glycosylation revealed that aberrant glycosylation is a universal feature in various steps of malignant transformation and tumour progression. Aberrant glycosylation is associated with poor survival, cancer progression, and metastasis, such as overexpression of Thomsen-nouvelle (Tn)-, T-, and sialyl-T (sT) antigens. Main carriers of sT- and sTn-antigens were identified as the mucin MUC1 and CD44v6. On the other hand, the Lewis antigens and their sialylated derives (Lex/sLex and Lea/sLeA) are the most prominent cancer-associated epitopes on both glycoproteins and glycolipids, since their overexpression is related to CRC malignant transformations and may lead to increased tumour cell adhesion and motility, thereby resulting in metastasis. The project aims to discover new potential biomarkers for CRC early detection and to evaluate the therapeutic potential of antibodies against CRC-associated antigens, namely against Lewis antigens and Thomsen-nouvelle (Tn)-, T-, and sialyl-T (sT) antigens. For the same reason glycoproteins, such as MUC1, CAE and CD44 will be studied for the development of novel monoclonal antibody production. After biomarker validation, hybridoma technology has been chosen to produce novel antibodies against, sLe and/or CD44, immunizing the mice directly with cancer cell lines proteins. The hybridoma against CD44 show staining against CD44 and sLex/a and further screening must be performed. The second hybridoma line against sLe antigens also shows positivity against different total cell lysate of CRC cells. Two clones have been selected for further characterization. The clones will be validated either for CRC early diagnosis or CRC treatment potential

L1cam as an e-selectin ligand in colon cancer

POCI-01-0145-FEDER-007728 ref. 140_596817822Metastasis is the main cause of death among colorectal cancer (CRC) patients. E-selectin and its carbohydrate ligands, including sialyl Lewis X (sLeX) antigen, are key players in the binding of circulating tumor cells to the endothelium, which is one of the major events leading to organ invasion. Nevertheless, the identity of the glycoprotein scaffolds presenting these glycans in CRC remains unclear. In this study, we firstly have characterized the glycoengineered cell line SW620 transfected with the fucosyltransferase 6 (FUT6) coding for the α1,3-fucosyltransferase 6 (FUT6), which is the main enzyme responsible for the synthesis of sLeX in CRC. The SW620FUT6 cell line expressed high levels of sLeX antigen and E-selectin ligands. Moreover, it displayed increased migration ability. E-selectin ligand glycoproteins were isolated from the SW620FUT6 cell line, identified by mass spectrometry, and validated by flow cytometry and Western blot (WB). The most prominent E-selectin ligand we identified was the neural cell adhesion molecule L1 (L1CAM). Previous studies have shown association of L1CAM with metastasis in cancer, thus the novel role as E-selectin counter-receptor contributes to understand the molecular mechanism involving L1CAM in metastasis formation.publishersversionpublishe

A School-Based Program to Promote Well-Being in Preadolescents: Results From a Cluster Quasi-Experimental Controlled Study

Diario della Salute [My Health Diary] is a school-based program designed to enhance the subjective well-being and health of 12- to 13-year-old students. We hypothesized that providing students with the social and emotional skills to fulfill their potential and deal with common developmental tasks of adolescence (e.g., onset of puberty, identity development, increased responsibilities and academic demands) would result in improved well-being and health. The program comprises five standardized interactive lessons concerning common psychosocial and health issues in adolescence, and two narrative booklets addressed to both students and their parents. We evaluated the effectiveness of the program in terms of the students' subjective well-being, aggressive behavior, and health behavior. Using a quasi-experimental study design, schools in the intervention group implemented the full program and those in the comparison group received their regular curriculum. We administered measures of the study's objectives both before and after program implementation. Statistical analyses accounted for within-school clustering, potential socioeconomic and demographic confounding, and pre-implementation levels of these measures. We sampled 62 schools and allocated 2630 students to either an intervention or comparison group. Sociodemographic characteristics and baseline outcomes were balanced across study groups. Unexpectedly, respondents in the intervention group had 0.38 greater mean adjusted score of the WHO/Europe Health Behaviour in School-Aged Children Symptom Checklist instrument than respondents in the comparison group, indicating a reduction in subjective well-being. We did not observe any program effects on aggressive and health behaviors. The apparent reduction in subjective well-being reflected by an increased perception of psychosomatic complaints is suggestive of either increased emotional competence or, potentially, iatrogenic program effects. While greater emotional competence is positively associated with well-being over the course of life, the program in its present form should not be disseminated due to the possibility of adverse unintended effects

Acute Delta Hepatitis in Italy spanning three decades (1991–2019): Evidence for the effectiveness of the hepatitis B vaccination campaign

Updated incidence data of acute Delta virus hepatitis (HDV) are lacking worldwide. Our aim was to evaluate incidence of and risk factors for acute HDV in Italy after the introduction of the compulsory vaccination against hepatitis B virus (HBV) in 1991. Data were obtained from the National Surveillance System of acute viral hepatitis (SEIEVA). Independent predictors of HDV were assessed by logistic-regression analysis. The incidence of acute HDV per 1-million population declined from 3.2 cases in 1987 to 0.04 in 2019, parallel to that of acute HBV per 100,000 from 10.0 to 0.39 cases during the same period. The median age of cases increased from 27 years in the decade 1991-1999 to 44 years in the decade 2010-2019 (p < .001). Over the same period, the male/female ratio decreased from 3.8 to 2.1, the proportion of coinfections increased from 55% to 75% (p = .003) and that of HBsAg positive acute hepatitis tested for by IgM anti-HDV linearly decreased from 50.1% to 34.1% (p < .001). People born abroad accounted for 24.6% of cases in 2004-2010 and 32.1% in 2011-2019. In the period 2010-2019, risky sexual behaviour (O.R. 4.2; 95%CI: 1.4-12.8) was the sole independent predictor of acute HDV; conversely intravenous drug use was no longer associated (O.R. 1.25; 95%CI: 0.15-10.22) with this. In conclusion, HBV vaccination was an effective measure to control acute HDV. Intravenous drug use is no longer an efficient mode of HDV spread. Testing for IgM-anti HDV is a grey area requiring alert. Acute HDV in foreigners should be monitored in the years to come

Social Media-Related Geographic Information in the Context of Strategic Environmental Assessment of Municipal Masterplans: A Case Study Concerning Sardinia (Italy)

This paper proposes a discussion concerning the use of social media-related geographic information in the context of the strategic environmental assessment (SEA) of Sardinian Municipal masterplans (MMPs). We show that this kind of information improves, substantially, the SEA process since it provides planners, evaluators, and the local communities with information retrieved from social media that would have not been available otherwise. This information integrates authoritative data collection, which comes from official sources, and enlightens tastes and preferences of the users of services and infrastructure, and their expectations concerning their spatial organization. A methodological approach related to the collection of social media-related geographic information is implemented and discussed with reference to the urban context of the city of Cagliari (Sardinia, Italy). The results are very effective in terms of provision of information, which may possibly increase the spatial knowledge available for planning policy definition and implementation. In this perspective, this kind of information discloses opportunities for building analytical scenarios related to urban and regional planning and it offers useful suggestions for sustainable development based on tourism strategies

Dual Inhibition of CDK4/6 and PI3K/AKT/mTOR Signaling Impairs Energy Metabolism in MPM Cancer Cells

Background: Malignant pleural mesothelioma (MPM) is an aggressive malignancy associated to asbestos exposure. One of the most frequent genetic alteration in MPM patients is CDKN2A/ARF loss, leading to aberrant activation of the Rb pathway. In MPM cells, we previously demonstrated the therapeutic efficacy of targeting this signaling with the CDK4/6 inhibitor palbociclib in combination with PI3K/mTOR inhibitors. Here, we investigated whether such combination may have an impact on cell energy metabolism. Methods: The study was performed in MPM cells of different histotypes; metabolic analyses were conducted by measuring GLUT-1 expression and glucose uptake/consumption, and by SeaHorse technologies. Results: MPM cell models differed for their ability to adapt to metabolic stress conditions, such as glucose starvation and hypoxia. Independently of these differences, combined treatments with palbociclib and PI3K/mTOR inhibitors inhibited cell proliferation more efficaciously than single agents. The drugs alone reduced glucose uptake/consumption as well as glycolysis, and their combination further enhanced these effects under both normoxic and hypoxic conditions. Moreover, the drug combinations significantly impaired mitochondrial respiration as compared with individual treatments. These metabolic effects were mediated by the concomitant inhibition of Rb/E2F/c-myc and PI3K/AKT/mTOR signaling. Conclusions: Dual blockade of glycolysis and respiration contributes to the anti-tumor efficacy of palbociclib-PI3K/mTOR inhibitors combination

CDK4/6 Inhibition Enhances the Efficacy of Standard Chemotherapy Treatment in Malignant Pleural Mesothelioma Cells

Simple Summary Malignant pleural mesothelioma (MPM) is an aggressive disease affecting the pleura, and its incidence is increasing worldwide. Currently, the recommended systemic therapy for MPM is cisplatin/pemetrexed or immunotherapy with nivolumab and ipilimumab. Unfortunately, the prognosis remains poor, and there is an urgent need for new and effective treatments. In this study we investigated the potential antitumoral effects of combining abemaciclib with the standard chemotherapy drugs cisplatin and pemetrexed. Background: The loss of the CDKN2A/ARF (cyclin-dependent kinase inhibitor 2A/alternative reading frame) gene is the most common alteration in malignant pleural mesothelioma (MPM), with an incidence of about 70%, thus representing a novel target for mesothelioma treatment. In the present study, we evaluated the antitumor potential of combining the standard chemotherapy regimen used for unresectable MPM with the CDK4/6 (cyclin-dependent kinase 4 or 6) inhibitor abemaciclib. Methods: Cell viability, cell death, senescence, and autophagy induction were evaluated in two MPM cell lines and in a primary MPM cell culture. Results: The simultaneous treatment of abemaciclib with cisplatin and pemetrexed showed a greater antiproliferative effect than chemotherapy alone, both in MPM cell lines and in primary cells. This combined treatment induced cellular senescence or autophagic cell death, depending on the cell type. More in detail, the induction of cellular senescence was related to the increased expression of p21, whereas autophagy induction was due to the impairment of the AKT/mTOR signaling. Notably, the effect of the combination was irreversible and no resumption in tumor cell proliferation was observed after drug withdrawal. Conclusion: Our results demonstrated the therapeutic potential of CDK4/6 inhibitors in combination with chemotherapy for the treatment of MPM and are consistent with the recent positive results in the MiST2 arm in abemaciclib-treated patients