A High-Content Small Molecule Screen Identifies Sensitivity of Glioblastoma Stem Cells to Inhibition of Polo-Like Kinase 1

Glioblastoma multiforme (GBM) is the most common primary brain cancer in adults and there are few effective treatments. GBMs contain cells with molecular and cellular characteristics of neural stem cells that drive tumour growth. Here we compare responses of human glioblastoma-derived neural stem (GNS) cells and genetically normal neural stem (NS) cells to a panel of 160 small molecule kinase inhibitors. We used live-cell imaging and high content image analysis tools and identified JNJ-10198409 (J101) as an agent that induces mitotic arrest at prometaphase in GNS cells but not NS cells. Antibody microarrays and kinase profiling suggested that J101 responses are triggered by suppression of the active phosphorylated form of polo-like kinase 1 (Plk1) (phospho T210), with resultant spindle defects and arrest at prometaphase. We found that potent and specific Plk1 inhibitors already in clinical development (BI 2536, BI 6727 and GSK 461364) phenocopied J101 and were selective against GNS cells. Using a porcine brain endothelial cell blood-brain barrier model we also observed that these compounds exhibited greater blood-brain barrier permeability in vitro than J101. Our analysis of mouse mutant NS cells (INK4a/ARF^−/−, or p53^−/−), as well as the acute genetic deletion of p53 from a conditional p53 floxed NS cell line, suggests that the sensitivity of GNS cells to BI 2536 or J101 may be explained by the lack of a p53-mediated compensatory pathway. Together these data indicate that GBM stem cells are acutely susceptible to proliferative disruption by Plk1 inhibitors and that such agents may have immediate therapeutic value.     

Subunit polypeptide composition of rubisco and the origin of allopolyploid Arabidopsis suecica (Brassicaceae)

The polypeptide composition of the large and small subunits of Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase) from Arabidopsis thaliana, A. suecica and Cardaminopsis arenosa have been studied by IEF (isoelectric focusing) analysis. The putative recent alopolyploid origin of A. suecica is supported. The chloroplast encoded large subunits served to identify solely A. thaliana as the maternal parent whereas the nuclear encoded small subunits indicate C. arenosa as the paternal species.     

Das pyroelektrische Verhalten und das permanente elektrische Moment von menschlichem und tierischem Sehnengewebe

Zusammenfassung Bei menschlichem und tierischem Sehnengewebe wurde pyroelektrisches Verhalten festgestellt. Pyroelektrisches Verhalten bedeutet, daß eine permanente makroskopische elektrische Polarisation und somit ein permanentes elektrisches Moment in Richtung der pyroelektrischen Achse vorhanden ist.Die pyroelektrische (und daher auch piezoelektrische) Achse verläuft in der Längsrichtung der Sehnenfasern bzw. ihrer Kollagenfibrillen. Sie stimmt überein mit der optischen Faserachse, in deren Verlauf positiv einachsige Doppelbrechung bekannt ist. Sehnen mit leidlicher Parallelausrichtung ihrer Faserbündel zeigen dementsprechend makroskopisch eine pyroelektrische Achse parallel zur physiologischen Sehnenlängsachse. Bei seilartiger Bündelung der Fibrillen oder anderen histologischen Unregelmäßigkeiten im Faserverlauf zeigen sich korrespondierende Unregelmäßigkeiten des pyroelektrischen Achsenverlaufs.In genau korrespondierenden Abschnitten der gleichen Sehne verschiedener Individuen fand sich eine sehr erhebliche biologische Streubreite der pyro- und piezoelektrischen Effekte.In situ hatte das permanente elektrische Moment der untersuchten menschlichen und Rinder-Sehnen, sowie der Schwanzsehnen von Ratten eine festliegende gleichsinnige Vektorrichtung bei allen Individuen. Die Vektorrichtung war bei den untersuchten korrespondierenden Sehnen von Mensch und Rind sowie bei den Rattenschwanz-Sehnen analog.Die Vektorrichtung der permanenten elektrischen Polarisation bei Sehnengewebe, und wahrscheinlich auch bei anderen kollagenhaltigen Strukturen, scheint in Beziehung zur morphogenetischen Entwicklungsrichtung der Tiere zu stehen.

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Summary Tendons are pyroelectric. Pyroelectrical behaviour is identical with a permanent electrical moment in the direction of the pyroelectrical axis. There exists a pyroelectrical (and consequently also piezoelectrical) axis corresponding to the longitudinal axis of the tendon fibres and their collagen fibrils, and also corresponding to their optical axis, in the direction of which a positive birefringence is known.Accordingly in tendons with an approximately parallel alignment of the collagen fibres the pyroelectrical (and piezoelectrical) axis corresponds to the physiological longitudinal tendon axis. In rope-like or any other histological irregular forms of the collagen fibre alignment corresponding irregularities of the electrical axis direction were found.The pyro- and piezoelectric effects show large biological variations in corresponding tendons of different individuals.In situ the permanent electrical moment in the examined tendons of men and ox and of rat-tail respectively is always in the same vector direction in each individual. This vector direction is analogical in all examined corresponding tendons of men, ox, and also in rat-tail. The permanent electrical moment of tendons (as well as of other collagen-containing tissues) seems to correspond to the morphogenetical development axis of animals.The results of this investigation may be understood by the assumption that the rod-like collagen molecule (i.e. tropocollagen) represents an electrical dipole with a permanent electrical moment in the longitudinal direction of the rod.

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Unterstützt durch Sachbeihilfen der Deutschen Forschungsgemeinschaft.     

Genic homozygosity in an ancient reptile (Alligator mississippiensis)

Proteins assayed electrophoretically showed variation at only three of 49 presumed genetic loci in alligators from southwestern Louisiana. Average heterozygosity per individual was 0.021±0.012; proportion of polymorphic loci was 0.06. Data on the history, structure, and ecology of this alligator population are consistent with natural selection as the primary factor accounting for this low genetic variability. However, neither a historic population bottleneck nor some genetic mechanism limiting variability can be dismissed as a possible factor.The study was supported by NSF Grant BMS 73-0125 to H.C.D.