Regulation der Genexpression von MYCN in humanen Neuoblastomzellen durch Transkriptionsfaktoren der E2F-Familie

Seit fast 30 Jahren ist bekannt, dass die Amplifikation und Expression des Onkogens MYCN in Neuroblastomen mit einer sehr ungünstigen Prognose für die Patienten einhergeht. Dennoch liegen die Mechanismen der Genregulation von MYCN weiterhin größtenteils im Dunkeln. Die Präsenz potentieller Bindungsstellen für E2F-Proteine im Promotor des MYCN-Gens sowie Zellkulturexperimente lieferten Hinweise auf eine Rolle der Transkriptionsfaktoren der E2F-Familie in der Regulation der N-myc-Expression. Ziel dieser Arbeit war die Beantwortung der Frage, ob E2F-Proteine notwendig sind, um eine primär hohe Expression von N-myc in Neuroblastomzellen mit Amplifikation des Onkogens aufrechtzuerhalten, und ob sie hinreichend sind, um die Transkription von MYCN in Zellen ohne endogene Expression von N-myc einzuleiten. Durch Überexpression des Tumorsuppressorproteins p16, welches zu einer Inaktivierung endogener E2F-Proteine führt, konnte die MYCN-mRNA-Menge in Neuroblastomzellen deutlich gesenkt werden. Vergleichbare Resultate wurden durch Expression von dominant negativem E2F-1 erzielt. Da in einigen Studien gezeigt werden konnte, dass Myc-Proteine ihrerseits E2F-Gene aktivieren können, nehmen wir an, in aggressiven Neuroblastomen könnte eine positive Rückkopplungsschleife zwischen E2F-Transkriptionsfaktoren auf der einen und N-myc auf der anderen Seite existieren, die die gesteigerte Aktivität des MYCN-Onkogens aufrechterhält. Stabil transfizierte E2F-ER-Fusionsproteine waren jedoch nicht in der Lage, das endogene MYCN-Gen in Neuroblastomzellen ohne Expression von N-myc anzuschalten. E2F-Proteine werden folglich für das volle Ausmaß der starken Expression von N-myc in Neuroblastomen benötigt, sind aber nicht ausreichend, um das Onkogen MYCN in Zellen ohne endogenes N-myc zu aktivieren. In der Zukunft könnte durch Verhinderung der Bindung von E2F-Proteinen an den MYCN-Promotor oder durch gentherapeutische Ansätze, die z.B. mittels viraler Infektion den Signalweg zwischen p16 und E2F rekonstruieren, die Expression von N-myc in Neuroblastomen gesenkt werden, so dass die Aggressivität der Tumore reduziert und die individuelle Prognose der Patienten verbessert werden könnte

OMERACT consensus-based operational definition of contextual factors in rheumatology clinical trials: A mixed methods study

Objectives: To develop an operational definition of contextual factors (CF) [1]. Methods: Based on previously conducted interviews, we presented three CF types in a Delphi survey; Effect Modifying -, Outcome Influencing - and Measurement Affecting CFs. Subsequently, a virtual Special Interest Group (SIG) session was held for in depth discussion of Effect Modifying CFs. Results: Of 161 Delphi participants, 129 (80%) completed both rounds. After two rounds, we reached consensus (≥70% agreeing) for all but two statements. The 45 SIG participants were broadly supportive. Conclusion: Through consensus we developed an operational definition of CFs, which was well received by OMERACT members

Identification of genetic variants associated with Huntington's disease progression: a genome-wide association study

Background Huntington's disease is caused by a CAG repeat expansion in the huntingtin gene, HTT. Age at onset has been used as a quantitative phenotype in genetic analysis looking for Huntington's disease modifiers, but is hard to define and not always available. Therefore, we aimed to generate a novel measure of disease progression and to identify genetic markers associated with this progression measure. Methods We generated a progression score on the basis of principal component analysis of prospectively acquired longitudinal changes in motor, cognitive, and imaging measures in the 218 indivduals in the TRACK-HD cohort of Huntington's disease gene mutation carriers (data collected 2008–11). We generated a parallel progression score using data from 1773 previously genotyped participants from the European Huntington's Disease Network REGISTRY study of Huntington's disease mutation carriers (data collected 2003–13). We did a genome-wide association analyses in terms of progression for 216 TRACK-HD participants and 1773 REGISTRY participants, then a meta-analysis of these results was undertaken. Findings Longitudinal motor, cognitive, and imaging scores were correlated with each other in TRACK-HD participants, justifying use of a single, cross-domain measure of disease progression in both studies. The TRACK-HD and REGISTRY progression measures were correlated with each other (r=0·674), and with age at onset (TRACK-HD, r=0·315; REGISTRY, r=0·234). The meta-analysis of progression in TRACK-HD and REGISTRY gave a genome-wide significant signal (p=1·12 × 10−10) on chromosome 5 spanning three genes: MSH3, DHFR, and MTRNR2L2. The genes in this locus were associated with progression in TRACK-HD (MSH3 p=2·94 × 10−8 DHFR p=8·37 × 10−7 MTRNR2L2 p=2·15 × 10−9) and to a lesser extent in REGISTRY (MSH3 p=9·36 × 10−4 DHFR p=8·45 × 10−4 MTRNR2L2 p=1·20 × 10−3). The lead single nucleotide polymorphism (SNP) in TRACK-HD (rs557874766) was genome-wide significant in the meta-analysis (p=1·58 × 10−8), and encodes an aminoacid change (Pro67Ala) in MSH3. In TRACK-HD, each copy of the minor allele at this SNP was associated with a 0·4 units per year (95% CI 0·16–0·66) reduction in the rate of change of the Unified Huntington's Disease Rating Scale (UHDRS) Total Motor Score, and a reduction of 0·12 units per year (95% CI 0·06–0·18) in the rate of change of UHDRS Total Functional Capacity score. These associations remained significant after adjusting for age of onset. Interpretation The multidomain progression measure in TRACK-HD was associated with a functional variant that was genome-wide significant in our meta-analysis. The association in only 216 participants implies that the progression measure is a sensitive reflection of disease burden, that the effect size at this locus is large, or both. Knockout of Msh3 reduces somatic expansion in Huntington's disease mouse models, suggesting this mechanism as an area for future therapeutic investigation

First imaging results from Apertif, a phased-array feed for WSRT

Apertif is a phased-array feed for the Westerbork Synthesis Radio Telescope (WSRT), increasing the field of view of the telescope by a factor of twenty-five. In 2017, three legacy surveys will commence: a shallow imaging survey, a medium-deep imaging survey, and a pulsars and fast transients survey. The medium-deep imaging survey will include coverage of the northern Herschel Atlas field, the CVn region, HetDex, and the Perseus-Pisces supercluster. The shallow imaging survey increases overlap with HetDex, has expanded coverage of the Perseus-Pisces supercluster, and includes part of the Zone of Avoidance. Both imaging surveys are coordinating with MaNGA and will have WEAVE follow-up. The imaging surveys will be done in full polarization over the frequency range 1130-1430 MHz, which corresponds to redshifts of z=0-0.256 for neutral hydrogen (HI). The spectral resolution is 12.2 kHz, or an HI velocity resolution of 2.6 km/s at z=0 and 3.2 km/s at z=0.256. The full resolution images will have a beam size of 15"x15"/sin(declination), and tapered data products (i.e., 30" resolution images) will also be available. The shallow survey will cover ~3500 square degrees with a four-sigma HI imaging sensitivity of 2.5x10^20 atoms cm^-2 (20 km/s linewidth) at the highest resolution and a continuum sensitivity of 15 uJy/beam (11 uJy/beam for polarization data). The current plan calls for the medium deep survey to cover 450 square degrees and provide an HI imaging sensitivity of 1.0x10^20 atoms cm^-2 at the highest resolution and a continuum sensitivity of 6 uJy/beam, close to the confusion limit (4 uJy/beam for polarization data, not confusion limited). Up-to-date information on Apertif and the planned surveys can be found at: http://www.apertif.nl.Commissioning of the Apertif instrument is currently underway. Here we present first results from the image commissioning, including the detection of HI absorption plus continuum and HI imaging. These results highlight the data quality that will be achieved for the surveys

The intermediate proteasome is constitutively expressed in pancreatic beta cells and upregulated by stimulatory, low concentrations of interleukin 1 β.

A central and still open question regarding the pathogenesis of autoimmune diseases, such as type 1 diabetes, concerns the processes that underlie the generation of MHC-presented autoantigenic epitopes that become targets of autoimmune attack. Proteasomal degradation is a key step in processing of proteins for MHC class I presentation. Different types of proteasomes can be expressed in cells dictating the repertoire of peptides presented by the MHC class I complex. Of particular interest for type 1 diabetes is the proteasomal configuration of pancreatic β cells, as this might facilitate autoantigen presentation by β cells and thereby their T-cell mediated destruction. Here we investigated whether so-called inducible subunits of the proteasome are constitutively expressed in β cells, regulated by inflammatory signals and participate in the formation of active intermediate or immuno-proteasomes. We show that inducible proteasomal subunits are constitutively expressed in human and rodent islets and an insulin-secreting cell-line. Moreover, the β5i subunit is incorporated into active intermediate proteasomes that are bound to 19S or 11S regulatory particles. Finally, inducible subunit expression along with increase in total proteasome activities are further upregulated by low concentrations of IL-1β stimulating proinsulin biosynthesis. These findings suggest that the β cell proteasomal repertoire is more diverse than assumed previously and may be highly responsive to a local inflammatory islet environment