Comparative Study of Spheroids (3D) and Monolayer Cultures (2D) for the In Vitro Assessment of Cytotoxicity Induced by the Mycotoxins Sterigmatocystin, Ochratoxin A and Patulin.

Mycotoxins are secondary metabolites produced by filamentous fungi associated with a variety of acute and chronic foodborne diseases. Current toxicology studies mainly rely on monolayer cell cultures and animal models, which are undeniably affected by several limitations. To bridge the gap between the current in vitro toxicology approach and the in vivo predictability of the data, we here investigated the cytotoxic effects induced by the mycotoxins sterigmatocystin (STE), ochratoxin A (OTA) and patulin (PAT) on different 2D and 3D cell cultures. We focused on human tumours (neuroblastoma SH-SY5Y cells and epithelial breast cancer MDA-MB-213 cells) and healthy cells (bone marrow-derived mesenchymal stem cells, BM-MSC, and umbilical vein endothelial cells, HUVECs). The cytotoxicity of STE, OTA, and PAT was determined after 24, 48 and 72 h of exposure using an ATP assay in both culture models. Three-dimensional spheroids' morphology was also analysed using the MATLAB-based open source software AnaSP 1.4 version. Our results highlight how each cell line and different culture models showed specific sensitivities, reinforcing the importance of using more complex models for toxicology studies and a multiple cell line approach for an improved and more comprehensive risk assessment.

The Growing Importance of Three-Dimensional Models and Microphysiological Systems in the Assessment of Mycotoxin Toxicity.

Current investigations in the field of toxicology mostly rely on 2D cell cultures and animal models. Although well-accepted, the traditional 2D cell-culture approach has evident drawbacks and is distant from the in vivo microenvironment. To overcome these limitations, increasing efforts have been made in the development of alternative models that can better recapitulate the in vivo architecture of tissues and organs. Even though the use of 3D cultures is gaining popularity, there are still open questions on their robustness and standardization. In this review, we discuss the current spheroid culture and organ-on-a-chip techniques as well as the main conceptual and technical considerations for the correct establishment of such models. For each system, the toxicological functional assays are then discussed, highlighting their major advantages, disadvantages, and limitations. Finally, a focus on the applications of 3D cell culture for mycotoxin toxicity assessments is provided. Given the known difficulties in defining the safety ranges of exposure for regulatory agency policies, we are confident that the application of alternative methods may greatly improve the overall risk assessment.

miR-210-3p enriched extracellular vesicles from hypoxic neuroblastoma cells stimulate migration and invasion of target cells.

BACKGROUND: Tumor hypoxia stimulates release of extracellular vesicles (EVs) that facilitate short- and long-range intercellular communication and metastatization. Albeit hypoxia and EVs release are known features of Neuroblastoma (NB), a metastasis-prone childhood malignancy of the sympathetic nervous system, whether hypoxic EVs can facilitate NB dissemination is unclear. METHODS: Here we isolated and characterized EVs from normoxic and hypoxic NB cell culture supernatants and performed microRNA (miRNA) cargo analysis to identify key mediators of EVs biological effects. We then validated if EVs promote pro-metastatic features both in vitro and in an in vivo zebrafish model. RESULTS: EVs from NB cells cultured at different oxygen tensions did not differ for type and abundance of surface markers nor for biophysical properties. However, EVs derived from hypoxic NB cells (hEVs) were more potent than their normoxic counterpart in inducing NB cells migration and colony formation. miR-210-3p was the most abundant miRNA in the cargo of hEVs; mechanistically, overexpression of miR-210-3p in normoxic EVs conferred them pro-metastatic features, whereas miR-210-3p silencing suppressed the metastatic ability of hypoxic EVs both in vitro and in vivo. CONCLUSION: Our data identify a role for hypoxic EVs and their miR-210-3p cargo enrichment in the cellular and microenvironmental changes favoring NB dissemination.

Efficacy and safety of nilotinib as frontline treatment in elderly (> 65 years) chronic myeloid leukemia patients outside clinical trials.

Here, we report real-world evidence on the safety and efficacy of nilotinib as a first-line treatment in elderly patients with chronic phase CML, treated in 18 Italian centers. Sixty patients aged > 65 years (median age 72 years (65-84)) were reported: 13 patients were older than 75 years. Comorbidities were recorded at baseline in 56/60 patients. At 3 months of treatment, all patients obtained complete hematological response (CHR), 43 (71.6%) an early molecular response (EMR), while 47 (78%) reached a complete cytogenetic response (CCyR). At last follow-up, 63.4% of patients still had a deep molecular response (MR4 or better), 21.6% reached MR3 as best response and 11.6% persisted without MR. Most patients (85%) started the treatment at the standard dose (300 mg BID), maintained at 3 months in 80% of patients and at 6 months in 89% of them. At the last median follow-up of 46.3 months, 15 patients discontinued definitively the treatment (8 due to side effects, 4 died for unrelated CML causes, 1 for failure, 2 were lost to follow-up). One patient entered in treatment-free remission. As to safety, 6 patients (10%) experienced cardiovascular events after a median time of 20.9 months from the start. Our data showed that nilotinib could be, as first-line treatment, effective and relatively safe even in elderly CML patients. In this setting, more data in the long term are needed about possible dose reduction to improve the tolerability, while maintaining the optimal molecular response.

Development of an in vitro neuroblastoma 3D model and its application for sterigmatocystin-induced cytotoxicity testing.

Given the increasing importance of establishing better risk assessments for mycotoxins, novel in vitro tools for the evaluation of their toxicity are mandatory. In this study, an in vitro 3D spheroid model from SH-SY5Y cells, a human neuroblastoma cell line, was developed, optimized and characterized to test the cytotoxic effects caused by the mycotoxin sterigmatocystin (STE). STE induced a concentration- and time-dependent cell viability decrease in spheroids. Spheroids displayed cell disaggregation after STE exposure, increasing in a dose-dependent manner and over time. STE also induced apoptosis as confirmed by immunofluorescence staining and Western blot. Following the decreased proliferation and increased apoptosis, STE cytostasis effects were observed by migration assays both in 2D and 3D cell culture. Increased ROS generation, as well as DNA damage were also observed. Taken together, these data highlight the cytotoxic properties of STE and suggest that cell culture models play a pivotal role in the toxicological risk assessment of mycotoxins. The evaluation of cytotoxicity in spheroids (3D) rather than monolayer cultures (2D) is expected to more accurately reflect in vivo-like cell behaviour.

Evaluation of soft skills among Italian Healthcare Rehabilitators: A cross sectional study.

BACKGROUND: Healthcare rehabilitator skills can be grouped into hard and soft skills. Hard skills are specific and teachable, which can be defined and measured, while soft skills are less tangible and more difficult to quantify. The aim of this study is to investigate the level of knowledge of soft skills among Italian healthcare rehabilitators, and how they were acquired. DESIGN AND METHODS: Two hundred healthcare rehabilitators, who worked in Southern Italy were enrolled from September 1st to October 31st 2017, and interviewed with Computer-Assisted-Web-Interview (CAWI) software, to assess their level of soft skills. RESULTS: Healthcare rehabilitators showed significant satisfaction with university education (59.5%), particularly for theoretical training (64%), while significant dissatisfaction was found for technical-practical training (63.5%), training in patients' family management (66.5%) and stages participation to improve soft skills (59%). Dissatisfied rehabilitators were found for university education of soft skills (59%), particularly for interpersonal relationships with patients family (66.5%) and technical-practical train in(63.5%). Women considered the training courses about soft skills acquisition more useful than men (43.8%). CONCLUSIONS: Healthcare rehabilitator training is lacking in the teaching of both technical-practical and soft skills. It is striking that in a healthcare profession like that of the rehabilitator, where practical and empathic skills are fundamental in the relationship with the patients, such skills are not treated in analogously with theoretical training.

Influenza vaccination and healthcare workers: barriers and predisposing factors.

BACKGROUND AND AIM OF THE WORK: Influenza is a disease that affects a large part of the world's population annually, with major health, social and economic impacts. Active immunisation practices have always been recommended to counter influenza, especially for people at risk. The recommendations of major health agencies strongly advise influenza vaccination for all healthcare workers, mostly for those in contact with at-risk or immunocompromised individuals. Yet, the influenza vaccination coverage among healthcare workers remains rather low worldwide. This review explore barriers and the facilitators of health care professional toward influenza's vaccination. METHODS: Narrative review  consulting the databases: PubMed, CINAHL by combining keywords health care worker, flu, influenza, vaccination, barrier, resistence, hesitangy, between November 2019 and February 2020 Results. From the 1031 records initially, twenty-two primary studies were included in this narrative review. Our results show that the identified facilitators are: desire for self-protection, protection for loved ones and community. Instead, the barriers to vaccination identified are: fear of contracting influenza from the vaccination itself; not considering themselves at risk; to believing believe that their immune system is capable of managing a trivial disease; disease considered trivial, laziness; false beliefs. DISCUSSION AND CONCLUSION: Adherence rate on influenza vaccination among health professionals is quite low. The interventions that make it "complex and traceable" flu vaccination refusal increase adherence to this type of vaccination. The results show that current vaccination campaigns do not increase the rate of adherence by healthcare workers. Identifying the predisposing factors and barriers to such vaccination can help to create, develop and test targeted educational programmes.

Verteporfin induces apoptosis and reduces the stem cell-like properties in Neuroblastoma tumour-initiating cells through inhibition of the YAP/TAZ pathway.

Neuroblastoma is an embryonal malignancy of early childhood arising from the embryonic sympatho-adrenal lineage of the neural crest. About half of all cases are currently classified as high-risk of disease recurrence, with an overall survival rate of less than 40% at 5 years despite intensive therapy. Recent studies on matched primary tumours and at the relapse revealed downregulation of genes transcriptionally silenced by YAP as significant association with neuroblastoma relapse. Here, we evaluated the pharmacological targeting of YAP/TAZ with the YAP/TAZ-TEAD inhibitor Verteporfin (VP) in Tumour Initiating Cells (TICs) derived from High-Risk Neuroblastoma patients. VP treatment suppresses YAP/TAZ expression, induces apoptosis and causes the re-organization of the cytoskeleton reducing cells migration and clonogenic ability. Moreover, VP reduces the percentage of side population cells and ABC transporters involved in drug resistance, and the percentage of stem cell subpopulations CD133+ and CD44+ of TICs. Finally, we demonstrated that VP sensitizes TICs to the standard drugs used for neuroblastoma therapy etoposide and cis-platin opening the way to use VP as drug repositioning candidate for recurrent neuroblastoma.

A Psychometric Validation of the Decisional Conflict Scale in Italian Cancer Patients Scheduled for Insertion of Central Venous Access Devices.

BACKGROUND/AIM: In oncological settings, high-quality decision-making takes place when an adaptive pattern of cognitive and behavioural processes occurs, potentially limiting post-decisional regret and leading to an increment of adherence to the final decision. An example of a choice that requires a patient's involvement in the decision-making during cancer treatment occurs when the insertion of Central Vascular Access Device (CVAD) is proposed for chemotherapy administration. The aim of the current study was to develop and evaluate the psychometric properties of an Italian version of the Decisional Conflict Scale (DCS), including its factorial structure and its accuracy in discriminating the level of uncertainty in a sample of cancer patients during their decision-making process for the insertion of a CVAD for intravenous (IV) chemotherapy administration. MATERIALS AND METHODS: The study included 264 cancer patients with different diagnoses. To test the structural and psychometric properties of the Italian version of the DCS (DCS-ITA), exploratory factorial analysis was conducted followed by traditional classical test theory assessments of internal reliability and criterion validity. RESULTS: The Italian version of the DCS (DCS-ITA) demonstrated good internal consistency, acceptable construct validity, which was tested with exploratory factorial analysis, and good criterion validity, demonstrated by the ability of the scale to differentiate between patients who declared themselves certain about their choice and patients expressing uncertainty about the choice to make. CONCLUSION: Overall, the results of the study showed that the DCS-ITA is a psychometrically sound instrument that easily discriminates between patients who are experiencing a decisional conflict and those who are not. The DCS-ITA can be used as a valid and easy-to-use tool for the screening of the decisional conflict in oncological settings.

An investigation into Video Game Addiction in Pre-Adolescents and Adolescents: A Cross-Sectional Study.

BACKGROUND AND OBJECTIVES: Game addiction is an emerging problem in public health. A gaming disorder is characterized by a pattern of persistent or recurrent gaming behavior. The behavioral pattern is severe enough to implicate a significant involvement of family, social, educational, professional, or other relationships. Therefore, greater attention needs to be paid to potential addictive behaviors in terms of video games in order to identify both pre-adolescents and adolescents at risk and to provide them with adequate assistance. Materials and Methods: A random sample of 622 students including pre-adolescents and adolescents were enrolled from September 1st to October 31th 2016, and the Game Addiction Scale (GAS) interview was used to identify pathological students with both Monothetic and Polythetic analysis. RESULTS: This study shows the presence of pathological students is equal to 1.93%, with 37.46% and 4.50% obtained with Monothetic and Polythetic analysis (global and partial), respectively. In our sample, the most frequent were students with a gaming time of 1 or 2 h, and students with a day gaming frequency of 1, 2, or 3 times a day. The items with more pathological students were Item 2 (i.e., Tolerance) and 4 (i.e., Withdrawal). Every item was positively correlated with Daily gaming time(hours) and Daily game frequency, excluding Item 4(i.e., Withdrawal). Finally, the Monothetic GAS score was positively correlated with Daily gaming time while the Polythetic Global GAS was positively correlated with Daily game frequency and negatively with Education level; instead, the Polythetic Partial GAS score was positively correlated with only Daily gaming time. CONCLUSION: Males are pathological gamblers more so than females and spend more time playing video games. An increase in Daily game frequency or Daily gaming time implicates an increase in video game addictions, while an increase in Education level, which generally corresponds to a greater age, implicates a decrease in game addiction. Finally, we observed that the correlations obtained between the Polythetic Partial GAS score with the independent variables such as Age, Gender, Education level, Daily gaming time (hours), and Daily game frequency were analogous to the significant correlations obtained with the Monothetic GAS score, while these correlations were different for the Polythetic Global GAS and the independent variables. These results suggest that the use of the original Polythetic scale should not be neglected.

An investigation on parenting stress of children with cystic fibrosis.

BACKGROUND: The management of chronic diseases, particularly in children, requires an integrated physical and psychological approach to both sick children and their family. This is the case of Cystic Fibrosis (CF), a complex genetic chronic disease, where, a comprehensive evaluation of the emotional impact and an effective multidimensional approach are indicated. AIM: This study investigates on parenting stress in children and adolescents with CF and its determinants related to parents, children and the disease severity. METHODS: The study involved 34.04% adult males and 65.96% adult females (range 21-55 years) and 47 children with CF, 54.35% males and 45.65% females (range 1-17 years). The data were obtained through a Parenting Stress Index - Short Form (PSI-SF) questionnaire. According to the PSI-SF scoring system, three types of stress were detected: a typical stress pattern (normal), a high stress pattern (increased) and a defensive response, which may be considered as a high stress feature in children which requires monitoring and clinical evaluation. RESULTS: This study shows a significant presence of stress in females (60.23%), of subject married (84.62%), unemployed (69.23%) and with education level such as "middle School" (61.54%). Concerning children of parents with high stress, it resulted most frequent children with one sibling (53.85%). Finally, by univariate analysis, it resulted a significant positive correlation between parenting stress and disease degree of children. Instead by multivariate analysis, we found that the variables: Number of siblings and Birth order were a significant positive and negative predictor of parenting stress respectively. CONCLUSION: An increased stress level was detected in less than one third of parents of subjects with CF. These data may be related to the psychological support which is part of the routine management of CF care team. However, as children's features seem to act as a determinant of stress more than parental ones, the parental-child dysfunction should be the target for further integrated interventions.

The CiTAS scale for evaluating taste alteration induced by chemotherapy: state of the art on its clinical use.

BACKGROUND: Cancer is the leading cause of death worldwide. Of the various therapeutic approaches, chemotherapy is the most widely used treatment. Among the various side effects associated with this treatment, taste alterations (TAs) have received little attention, even though they have a serious impact on the nutritional aspect and quality of life (QoL) of patients. TAs concern 75% of the patients receiving chemotherapy, and this figure is still considered to be underestimatedbeacuse could be due both to inadequate attention and to the absence of specific subjective tools able to fully evaluate TAs in patients undergoing chemotherapy. METHODS: A review by querying CINAHL, PubMed, Scopus and Google Scholar databases about the current status of use of the CiTAS self-evaluation scale, was performed. RESULTS: From critical reading of the selected reports, it can be said that until now CiTAS has not been used to a large extent for evaluating taste, even at a late stage in patients undergoing chemotherapy. However, the results and the selected reports seem to indicate hope for its wider use. CONCLUSIONS: In literature, CITAS scale has been used on very heterogeneous populations and not adequately studied in specific care settings, its use within controlled trials could implement its spread.Correct and subjective evaluation of TAs would allow the planning of specific and personalized interventions aimed at providing adequate nutrition to support the maintenance and/or achievement of a correct body mass index. All this could contribute significantly to a better perception of QoL in patients undergoing chemotherapy.

Cornell Assessment of Pediatric Delirium: Italian cultural validation and preliminary testing.

INTRODUCTION: Paediatric delirium is associated with a longer duration of hospitalization in paediatric intensive care units, the emergence of post-traumatic symptoms and possible neurocognitive dysfunction after discharge. In preschool children, the diagnosis of delirium appears rather challenging: their pre-verbal status and the presence of cognitive skills still in development make accurate diagnosis difficult. Recently, a pediatric delirium screening tool suitable for critical preschool children has also been developed and identified in international literature, with excellent results also in critical infants under 2 years of age: the Cornell Assessment of Pediatric Delirium (CAPD). The CAPD, using a Likert scale, bases the assessment of paediatric delirium within the context of child development. This scale follows the development of the infant by comparing the detection of specific items on the scale as the anchor points that characterize the development of infants by age groups. OBJECTIVE: Culturally and linguistically validation in Italian language and prior testing of the Cornell Assessment of Pediatric Delirium. METHOD: Translation and Cultural Validation of the Cornell Assessment of Pediatric Delirium (CAPD) for the Evaluation/Diagnosis of Pediatric Delirium within Pediatric Intensive Care. The translation and adaptation of this instrument followed the phases of the model proposed by the World Health Organization. Prior testings, such as item descriptive analysis, item-total correlation and Cronbach's alpha, were conducted. RESULTS: All phases of the cultural-linguistic validation process were carried out in a satisfactory manner. For the prior testing, the scale was administered to a sample of 42 children, with age ranged 0-5 years old (66.6%), with a higher prevalence of the male gender. All items were normally distributed and there was no excessive Skeweness and Kurtosis. Each item contributed to the scale fairly well and all coefficients of item total correlation (rjx) were higher than the recommended level of 0.30. The composite reliability index was 0.94 and Cronbach's alpha was 0.96. CONCLUSIONS: The process has meticulously followed the recommendations in international literature. The final version was approved by the authors of the original instrument.

Human Papillomavirus Infection and Vaccination: Knowledge and Attitudes among Nursing Students in Italy.

This cross-sectional study assessed nursing students' knowledge and attitudes about Human papillomavirus (HPV) infection and vaccination in Italy. The survey was conducted among a sample of 556 nursing students. Almost all reported that they had heard about HPV infection, while only 36.5% knew the risk factors of HPV infection and that this could be prevented by the HPV vaccine. Those who had heard about HPV infection during their degree program were more likely to know risk factors of HPV infection and that this could be prevented by the HPV vaccine. The majority of students (65.3%) reported that they would be willing to receive the HPV vaccine. Moreover, 91.7% of participants reported that they were willing, as future health care operators, to recommend the HPV vaccine to others. Those who knew risk factors of HPV infection and that this could be prevented by the HPV vaccine, and those who knew that cervical cancer could be prevented by the HPV vaccine expressed this positive attitude about willingness to recommend the HPV vaccine. These results highlight the need to supplement nursing students' specific education, to improve their knowledge and awareness of HPV vaccination.

Chromosome instability in neuroblastoma.

Neuroblastoma is a neural crest-derived tumor that accounts for 7-10% of all malignancies in children and ~15% of all childhood cancer-associated mortalities. Approximately 50% of patients are characterized as high-risk (HR) and have an overall survival of <40% at 5 years from diagnosis. HR patients with unfavorable prognosis exhibit several structural copy number variations (CNVs), whereas localized tumors belonging to patients in the low- and intermediate-risk classes, have favorable outcomes and display several numerical CNVs. Taken together these results are indicative of chromosome instability (CIN) in neuroblastoma tumor cells. The present review discusses multiple aspects of CIN including methods of measuring CIN, CIN targeting as a therapeutic strategy in cancer and the effects of CIN in neuroblastoma development and aggressiveness with particular emphasis on the CIN gene signature associated with HR neuroblastoma patients.

[Effectiveness of the Situation-Background-Assessment-Recommendation (SBAR) methodology in patient handovers between nurses, Italy]. / Implementazione nella sicurezza delle cure e la continuità assistenziale: revisione narrativa della letteratura sull'efficacia del metodo Sbar.

Exchange of information between healthcare professionals involved in the care of patients is essential for providing effective care. Use of the Situation-BackgroundAssessment-Recommendation (SBAR) methodology is increasingly indicated in the health sector to facilitate communication during handovers of patients between clinicians or clinical teams. The present review of the literature aims to determine the effectiveness of the SBAR methodology in the handover of nursing documentation.

Somatic mutations in specific and connected subpathways are associated with short neuroblastoma patients' survival and indicate proteins targetable at onset of disease.

Neuroblastoma (NB) is an embryonic malignancy of the sympathetic nervous system with heterogeneous biological, morphological, genetic and clinical characteristics. Although genomic studies revealed the specific biological features of NB pathogenesis useful for new therapeutic approaches, the improvement of high-risk (HR)-NB patients overall survival remains unsatisfactory. To further clarify the biological basis of disease aggressiveness, we used whole-exome sequencing to examine the genomic landscape of HR-NB patients at stage M with short survival (SS) and long survival (LS). Only a few genes, including SMARCA4, SMO, ZNF44 and CHD2, were recurrently and specifically mutated in the SS group, confirming the low recurrence of common mutations in this tumor. A systems biology approach revealed that in the two patient groups, mutations occurred in different pathways. Mutated genes (ARHGEF11, CACNA1G, FGF4, PTPRA, PTK2, ANK3, SMO, NTNG2, VCL and NID2) regulate the MAPK pathway associated with the organization of the extracellular matrix, cell motility through PTK2 signaling and matrix metalloproteinase activity. Moreover, we detected mutations in LAMA2, PTK2, LAMA4, and MMP14 genes, impairing MET signaling, in SFI1 and CHD2 involved in centrosome maturation and chromosome remodeling, in AK7 and SPTLC2, which regulate the metabolism of nucleotides and lipoproteins, and in NALCN, SLC12A1, SLC9A9, which are involved in the transport of small molecules. Notably, connected networks of somatically mutated genes specific for SS patients were identified. The detection of mutated genes present at the onset of disease may help to address an early treatment of HR-NB patients using FDA-approved compounds targeting the deregulated pathways.

Knowledge, attitudes, and practice on the prevention of central line-associated bloodstream infections among nurses in oncological care: A cross-sectional study in an area of southern Italy.

The objectives of the cross-sectional study were to delineate the knowledge, attitudes, and behavior among nurses regarding the prevention of central line-associated bloodstream infections (CLABSIs) and to identify their predisposing factors. A questionnaire was self-administered from September to November 2011 to nurses in oncology and outpatient chemotherapy units in 16 teaching and non-teaching public and private hospitals in the Campania region (Italy). The questionnaire gathered information on demographic and occupational characteristics; knowledge about evidence-based practices for the prevention of CLABSIs; attitudes towards guidelines, the risk of transmitting infections, and hand-washing when using central venous catheter (CVC); practices about catheter site care; and sources of information. The vast majority of the 335 nurses answered questions correctly about the main recommendations to prevent CLABSIs (use sterile gauze or sterile transparent semipermeable dressing to cover the catheter site, disinfect the needleless connectors before administer medication or fluid, disinfect with hydrogen peroxide the catheter insertion site, and use routinely anticoagulants solutions). Nurses aged 36 to 50 years were less likely to know these main recommendations to prevent CLABSIs, whereas this knowledge was higher in those who have received information about the prevention of these infections from courses. Nurses with lower education and those who do not know two of the main recommendations on the site's care to prevent the CLABSIs, were more likely to perceive the risk of transmitting an infection. Higher education, attitude toward the utility allow to dry antiseptic, and the need of washing hands before wearing gloves for access to port infusion were predictors of performing skin antiseptic and aseptic technique for dressing the catheter insertion site. Educational interventions should be implemented to address the gaps regarding knowledge and practice regarding the prevention of CLABSIs and to ensure that nurses use evidence-based prevention interventions.

Neuroblastoma treatment in the post-genomic era.

Neuroblastoma is an embryonic malignancy of early childhood originating from neural crest cells and showing heterogeneous biological, morphological, genetic and clinical characteristics. The correct stratification of neuroblastoma patients within risk groups (low, intermediate, high and ultra-high) is critical for the adequate treatment of the patients.High-throughput technologies in the Omics disciplines are leading to significant insights into the molecular pathogenesis of neuroblastoma. Nonetheless, further study of Omics data is necessary to better characterise neuroblastoma tumour biology. In the present review, we report an update of compounds that are used in preclinical tests and/or in Phase I-II trials for neuroblastoma. Furthermore, we recapitulate a number of compounds targeting proteins associated to neuroblastoma: MYCN (direct and indirect inhibitors) and downstream targets, Trk, ALK and its downstream signalling pathways. In particular, for the latter, given the frequency of ALK gene deregulation in neuroblastoma patients, we discuss on second-generation ALK inhibitors in preclinical or clinical phases developed for the treatment of neuroblastoma patients resistant to crizotinib.We summarise how Omics drive clinical trials for neuroblastoma treatment and how much the research of biological targets is useful for personalised medicine. Finally, we give an overview of the most recent druggable targets selected by Omics investigation and discuss how the Omics results can provide us additional advantages for overcoming tumour drug resistance.

Exome and deep sequencing of clinically aggressive neuroblastoma reveal somatic mutations that affect key pathways involved in cancer progression.

The spectrum of somatic mutation of the most aggressive forms of neuroblastoma is not completely determined. We sought to identify potential cancer drivers in clinically aggressive neuroblastoma.Whole exome sequencing was conducted on 17 germline and tumor DNA samples from high-risk patients with adverse events within 36 months from diagnosis (HR-Event3) to identify somatic mutations and deep targeted sequencing of 134 genes selected from the initial screening in additional 48 germline and tumor pairs (62.5% HR-Event3 and high-risk patients), 17 HR-Event3 tumors and 17 human-derived neuroblastoma cell lines.We revealed 22 significantly mutated genes, many of which implicated in cancer progression. Fifteen genes (68.2%) were highly expressed in neuroblastoma supporting their involvement in the disease. CHD9, a cancer driver gene, was the most significantly altered (4.0% of cases) after ALK.Other genes (PTK2, NAV3, NAV1, FZD1 and ATRX), expressed in neuroblastoma and involved in cell invasion and migration were mutated at frequency ranged from 4% to 2%.Focal adhesion and regulation of actin cytoskeleton pathways, were frequently disrupted (14.1% of cases) thus suggesting potential novel therapeutic strategies to prevent disease progression.Notably BARD1, CHEK2 and AXIN2 were enriched in rare, potentially pathogenic, germline variants.In summary, whole exome and deep targeted sequencing identified novel cancer genes of clinically aggressive neuroblastoma. Our analyses show pathway-level implications of infrequently mutated genes in leading neuroblastoma progression.