Common genetic variants in NEFL influence gene expression and neuroblastoma risk

The genetic etiology of sporadic neuroblastoma is still largely obscure. In a genome-wide association study, we identified single-nucleotide polymorphisms (SNP) associated with neuroblastoma at the CASC15, BARD1, LMO1, DUSP12, HSD17B12, HACE1, and LIN28B gene loci, but these explain only a small fraction of neuroblastoma heritability. Other neuroblastoma susceptibility genes are likely hidden among signals discarded by the multiple testing corrections. In this study, we evaluated eight additional genes selected as candidates for further study based on proven involvement in neuroblastoma differentiation. SNPs at these candidate genes were tested for association with disease susceptibility in 2,101 cases and 4,202 controls, with the associations found replicated in an independent cohort of 459 cases and 809 controls. Replicated associations were further studied for cis-effect using gene expression, transient overexpression, silencing, and cellular differentiation assays. The neurofilament gene NEFL harbored three SNPs associated with neuroblastoma (rs11994014: Pcombined \ubc 0.0050; OR, 0.88; rs2979704: Pcombined \ubc 0.0072; OR, 0.87; rs1059111: Pcombined \ubc 0.0049; OR, 0.86). The protective allele of rs1059111 correlated with increased NEFL expression. Biologic investigations showed that ectopic overexpression of NEFL inhibited cell growth specifically in neuroblastoma cells carrying the protective allele. NEFL overexpression also enhanced differentiation and impaired the proliferation and anchorage-independent growth of cells with protective allele and basal NEFL expression, while impairing invasiveness and proliferation of cells homozygous for the risk genotype. Clinically, high levels of NEFL expression in primary neuroblastoma specimens were associated with better overall survival (P \ubc 0.03; HR, 0.68). Our results show that common variants of NEFL influence neuroblastoma susceptibility and they establish that NEFL expression influences disease initiation and progressio

Cellular proteome alterations in response to hypoxia inducible factor HIF-2\u3b1 in normoxic neuroblastoma cells

Il neuroblastoma (NB) \ue8 un tumore embrionale del sistema nervoso simpatico, che deriva da cellule della cresta neurale. \uc8 il tumore extracranico pi\uf9 diffuso tra i bambini di et\ue0 inferiore a un anno e rappresenta circa il 7% di tutti i tumori infantili. L'ipossia si sviluppa comunemente durante la crescita tumorale ed \ue8 associata ad una prognosi infausta con resistenza ai trattamenti terapeutici. Molte evidenze suggeriscono una correlazione tra i fattori ipossia-inducibile (HIF), HIF-1\u3b1 e HIF-2\u3b1, con il grado di differenziamento e quindi l\u2019aggressivit\ue0 tumorale. In particolare, nel Neuroblastoma HIF-2\u3b1 \ue8 stabile anche in condizioni di normossiche e continua ad essere attivo anche dopo 48-72 ore dall\u2019ipossia. In NB HIF-2\u3b1, localizzato principalmente nelle nicchie peri-vascolare del tumore ed \ue8 correlato ad una prognosi infausta. Studi recenti hanno dimostrato il coinvolgimento di HIF-2\u3b1 nella proliferazione e nell\u2019aumento della aggressivit\ue0 tumorale. Lo scopo del progetto \ue8 quello di acquisire ulteriori informazioni sui meccanismi molecolari indotti dall\u2019over-espressione di HIF-2\u3b1 che sono alla base della resistenza ai convenzionali trattamenti terapeutici. Il nostro progetto, quindi, propone un approccio proteomico, basato sull\u2019analisi DIGE e sull\u2019identificazione di antigeni di membrana (FACS), per identificare nuovi bersagli prognostici e terapeutici per il trattamento clinico di forme di NB pi\uf9 aggressive e resistenti ai protocolli terapeutici convenzionali. Attraverso l\u2019indagine DIGE e studi funzionali sono stati identificati nel nostro sistema sperimentale diverse proteine coinvolte nel metabolismo cellulare e nei processi di regolazione dell\u2019mRNA. Inoltre sono stati identificati nuovi antigeni di membrana differenzialmente espressi soffermandoci sulle propriet\ue0 antiadesive e proinvasive di CD55, marker specifico di HIF-2\u3b1.Quindi, CD55 potrebbe essere usato nella diagnosi e per la stratificazione di pazienti affetti da forme pi\uf9 aggressive di Neuroblastoma.Neuroblastoma (NB) is an embryonal tumor of neuroectodermal cells derived from precursor or immature cells of the sympathetic nervous system (SNS). This disease rappresents the most common extracranial tumor in infants, accounting for 8% to 10% of all childhood cancer and for approximately 15% of cancer deaths in children. The deep knowledge of NB biology is imperative toward the development of novel therapy. Hypoxia is a typical feature of several solid tumors microenvironment and is associated with a poor prognosis and resistance to therapy. The relationship among hypoxia, tumor phenotypes and clinical parameters in NB is not well characterized. Tumor adaptation to hypoxia is mainly mediated by two transcription factors: the hypoxia-inducible factors (HIFs) HIF-1\u3b1 and HIF-2\u3b1. HIF-2\u3b1 is stable also in normoxia condition and continues to be active even after 48\u201372 h of hypoxia in some neuroblastoma cell lines thus indicate that HIF-2\u3b1 plays a critical role in driving the hypoxic response. Interesting, HIF-2\u3b1 is correlated with poor patient prognosis in NB and is localizated in tumor peri-vascular niches. These findings indicate that HIF-2\u3b1 protein expression in NB samples at normoxic levels might affect the aggressive tumor phenotype. The main aim of my phD program has been to get new insights into the molecular mechanism of tumor aggressiveness mediated by HIF-2\u3b1 protein overexpression in NB cells. Interesting, HIF-2\u3b1 overexpressing cells acquire an undifferentiated phenotype and the ability to grow as neurospheres in soft agar. Then I applied two different proteomic approaches, DIGE analysis and FACS detection of membrane antigens to identify new putative prognostic and therapeutic hypoxia-related targets to be used in clinical treatment of aggressive NB forms. The identified proteins have important roles in a variety of pathways such as \u201ccitrate cycle\u201d, \u201cglycolysis\u201d and \u201csplicesoma\u201d thus indicating that HIF-2\u3b1 over-expression affects the cellular metabolic balance and increases the processes of mRNA regulation. These findings might provide an innovative therapeutic strategy by combining anti-metabolic drugs and pathways inhibitors.Among the cell surface antigens which were differentialy HIF-2\u3b1 regulated CD55 was the most significantly expressed marker in our cellular system. I assessed CD55 has anti-adhesive and pro-invading functions that might provide the basis for NB solid tumors to survive as microscopic residual disease. Furthermore, the use of CD55 antibody-based visualization as in PET (Positron Emission Tomography) imaging will have implications for the development of more accurate diagnosis and prognosis in challenging cases and for driving personalized treatment. In conclusion, the HIF-2\u3b1 novel markers identified in this study might improve patients risk stratification and could be also used as putative drug targets being immunotherapy is one of the most promising anticancer treatment

Revues : outils et objets de l’histoire de l’art

Journals : tools and objects of art history Journals are the material support of art history, but they can also provide, in turn, a fascinating object for art historical inquiry : they are extremely useful tools for the study of art and its discourses. The present article, the result of collaborative research conducted at the INHA on francophone journals of art history and criticism (first half of the 20th century), presents three possible ways to rethink this field of inquiry : the history of journals as works of art in their own right, to be analyzed in both their aesthetic and material dimensions ; the history of the intellectual content of art history as a scientific discipline and its evolution over time ; and the social history of discourses on art, with the goal of defining the intellectual spaces in which positions are rooted and from which they issue, the forms of sociability and the publics they imply. By outlining the contours of this open-ended and prospective field within art history, we have also attempted to provide an overview of the methodological tools and bibliographic resources available for this kind of research.Objet à part entière de l’histoire de l’art, les revues en constituent également le support. Elles s’avèrent donc un outil particulièrement précieux à l’étude de l’art et de ses discours. Issu d’une réflexion collective sur les revues francophones d’histoire et de critique d’art de la première partie du XXe siècle menée à l’INHA, le présent article propose trois pistes pour repenser ce champ : l’histoire de la revue comme œuvre, à étudier dans ses dimensions esthétiques et matérielles ; l’histoire des contenus intellectuels de la discipline et leurs évolutions ; et l’histoire sociale du discours sur l’art, visant à cerner ses lieux d’implantation et d’énonciation, ses formes de sociabilité et ses publics. En présentant ce domaine ouvert et prospectif de notre discipline, nous avons également souhaité proposer un bilan des outils méthodologiques et bibliographiques disponibles.Fravalo Fabienne, Froissart Pezone Rossella, Karp Lugo Laura, Lafont Anne, Passini Michela, Piccioni Lucia. Revues : outils et objets de l’histoire de l’art. In: Histoire de l'art, N°68, 2011. Les enjeux de la restauration. pp. 121-131

Inhibition of hypoxia inducible factors combined with all-trans retinoic acid treatment enhances glial transdifferentiation of neuroblastoma cells

Neuroblastoma (NBL) is a heterogeneous tumor characterized by a wide range of clinical manifestations. A high tumor cell differentiation grade correlates to a favorable stage and positive outcome. Expression of the hypoxia inducible factors HIF1-α (HIF1A gene) and HIF2-α (EPAS1 gene) and/or hypoxia-regulated pathways has been shown to promote the undifferentiated phenotype of NBL cells. Our hypothesis is that HIF1A and EPAS1 expression represent one of the mechanisms responsible for the lack of responsiveness of NBL to differentiation therapy. Clinically, high levels of HIF1A and EPAS1 expression were associated with inferior survival in two NBL microarray datasets, and patient subgroups with lower expression of HIF1A and EPAS1 showed significant enrichment of pathways related to neuronal differentiation. In NBL cell lines, the combination of all-trans retinoic acid (ATRA) with HIF1A or EPAS1 silencing led to an acquired glial-cell phenotype and enhanced expression of glial-cell differentiation markers. Furthermore, HIF1A or EPAS1 silencing might promote cell senescence independent of ATRA treatment. Taken together, our data suggest that HIF inhibition coupled with ATRA treatment promotes differentiation into a more benign phenotype and cell senescence in vitro. These findings open the way for additional lines of attack in the treatment of NBL minimal residue disease

Association Study Between Coronary Artery Disease and rs1333049 Polymorphism at 9p21.3 Locus in Italian Population

In this study, we verify the association between the rs1333049 single nucleotide polymorphism (9p21.3) within CDKN2A-CDKN2B and coronary artery disease (CAD) in an Italian population. We replicated rs1333049_G allele association with a significantly reduced risk of CAD (OR = 0.816; 95% confidence interval [0.705â\u80\u930.945]; p = 0.0065) in 711 CAD patients and 755 normal healthy individuals. This effect is maintained even stratifying patients by gender and by risk factors. A significant association was found with age of CAD onset. Interestingly, we found a protective trend of association between the rs1333049_G allele and peripheral artery disease, a progressive atherosclerotic condition in which plaque builds up in the arteries that carry blood to the head, organs, and limbs (OR = 0.724; 95% CI [0.520â\u80\u931.007]; p = 0.054). No genotype-phenotype association was found with more severe CAD clinical parameters. If certain genetic factors predispose individuals to adverse outcomes, the knowledge of a patientâ\u80\u99s genotype may influence clinical management

HIF-1 transcription activity: HIF1A driven response in normoxia and in hypoxia

Abstract Background HIF1A (Hypoxia-Inducible-Factor 1A) expression in solid tumors is relevant to establish resistance to therapeutic approaches. The use of compounds direct against hypoxia signaling and HIF1A does not show clinical efficiency because of changeable oxygen concentrations in solid tumor areas. The identification of HIF1A targets expressed in both normoxia and hypoxia and of HIF1A/hypoxia signatures might meliorate the prognostic stratification and therapeutic successes in patients with high-risk solid tumors. Methods In this study, we conducted a combined analysis of RNA expression and DNA methylation of neuroblastoma cells silenced or unsilenced for HIF1A expression, grown in normoxia and hypoxia conditions. Results The analysis of pathways highlights HIF-1 (heterodimeric transcription factor 1) activity in normoxia in metabolic process and HIF-1 activity in hypoxia in neuronal differentiation process. HIF1A driven transcriptional response in hypoxia depends on epigenetic control at DNA methylation status of gene regulatory regions. Furthermore, low oxygen levels generate HIF1A-dependent or HIF1A-independent signatures, able to stratify patients according to risk categories. Conclusions These findings may help to understand the molecular mechanisms by which low oxygen levels reshape gene signatures and provide new direction for hypoxia targeting in solid tumor

Chromosomal Microarray Analysis versus Karyotyping in Fetuses with Increased Nuchal Translucency

We have carried out a retrospective study of chromosome anomalies associated with increased nuchal translucency (NT) in order to compare yield rates of karyotype, chromosome microarray analysis (CMA), and non-invasive prenatal testing (NIPT) in this condition. Presenting with increased NT or cystic hygroma ≥3.5 mm as an isolated sign, 249 fetuses underwent karyotype and/or CMA from 11 to 18 gestational weeks. Karyotype and fluorescence in situ hybridization (FISH) analyses detected 103 chromosomal anomalies including 95 aneuploidies and eight chromosomal rearrangements or derivatives. Further, seven pathogenic copy number variants (CNV), five likely pathogenic CNVs, and 15 variants of unknown significance (VOUS) were detected by CMA in fetuses with normal karyotype. Genetic testing is now facing new challenges due to results with uncertain clinical impacts. Additional investigations will be necessary to interpret these findings. More than 15% of the anomalies that we have diagnosed with invasive techniques could not be detected by NIPT. It is therefore definitely not recommended in the case of ultrasound anomalies. These results, while corroborating the use of CMA in fetuses with increased NT as a second tier after rapid aneuploidy testing, do not suggest a dismissal of karyotype analysis