Diffuse glioneuronal tumour with oligodendroglioma-like features and nuclear clusters (DGONC) - a molecularly defined glioneuronal CNS tumour class displaying recurrent monosomy 14.

AIMS: DNA methylation-based central nervous system (CNS) tumour classification has identified numerous molecularly distinct tumour types, and clinically relevant subgroups among known CNS tumour entities that were previously thought to represent homogeneous diseases. Our study aimed at characterizing a novel, molecularly defined variant of glioneuronal CNS tumour. PATIENTS AND METHODS: DNA methylation profiling was performed using the Infinium MethylationEPIC or 450 k BeadChip arrays (Illumina) and analysed using the 'conumee' package in R computing environment. Additional gene panel sequencing was also performed. Tumour samples were collected at the German Cancer Research Centre (DKFZ) and provided by multinational collaborators. Histological sections were also collected and independently reviewed. RESULTS: Genome-wide DNA methylation data from >25 000 CNS tumours were screened for clusters separated from established DNA methylation classes, revealing a novel group comprising 31 tumours, mainly found in paediatric patients. This DNA methylation-defined variant of low-grade CNS tumours with glioneuronal differentiation displays recurrent monosomy 14, nuclear clusters within a morphology that is otherwise reminiscent of oligodendroglioma and other established entities with clear cell histology, and a lack of genetic alterations commonly observed in other (paediatric) glioneuronal entities. CONCLUSIONS: DNA methylation-based tumour classification is an objective method of assessing tumour origins, which may aid in diagnosis, especially for atypical cases. With increasing sample size, methylation analysis allows for the identification of rare, putative new tumour entities, which are currently not recognized by the WHO classification. Our study revealed the existence of a DNA methylation-defined class of low-grade glioneuronal tumours with recurrent monosomy 14, oligodendroglioma-like features and nuclear clusters.

Recruitment of neural precursor cells from circumventricular organs of patients with cerebral ischaemia.

AIMS: Adult neurogenesis is well described in the subventricular zone of the lateral ventricle walls and in the subgranular zone of the hippocampal dentate gyrus. However, recent studies indicate that self-renewal of neural stem cells (NSCs) is not restricted to these niches, but that diverse areas of the adult brain are capable of generating new neurones and responding to various pathological alterations. In particular, NSCs have been identified in circumventricular organs (CVOs) of the adult mouse brain. METHODS: In order to detect possible neural stem or progenitor cells in CVOs of the human brain, we analysed post mortem human brain tissue from patients without neuropathological changes (n = 16) and brains from patients with ischaemic stroke (n = 16). RESULTS: In all analysed CVOs (area postrema, median eminence, pineal gland and neurohypophysis) we observed cells with expression of early NSC markers, such as GFAP, nestin, vimentin, OLIG2 and PSA-NCAM, with some of them coexpressing Ki67 as a marker of cell proliferation. Importantly, stroke patients displayed an up to fivefold increase with respect to the relative number of Ki67- and OLIG2-expressing cells within their CVOs. CONCLUSIONS: Our findings are compatible with a scenario where CVOs may serve as a further source of NSCs in the adult human brain and may contribute to neurogenesis and brain plasticity in the context of brain injury.

Expression of BARHL1 in medulloblastoma is associated with prolonged survival in mice and humans.

Medulloblastoma is the most common malignant brain tumor in childhood, and development of targeted therapies is highly desired. Although the molecular mechanisms of malignant transformation are not fully understood, it is known that medulloblastomas may arise from cerebellar granule neuron precursors. The homeodomain transcription factor Barhl1 is known to regulate migration and survival of granule cell precursors, but its functional role in medulloblastoma is unknown. We show here that the expression of BARHL1 is significantly upregulated during human cerebellar development and in human medulloblastoma samples as compared with the normal adult cerebellum. We also detected high levels of Barhl1 expression in medulloblastomas of Math1-cre:SmoM2 mice, a mouse model for Sonic hedgehog-associated medulloblastomas that we developed previously. To investigate Barhl1 function in vivo during tumor development, we generated Barhl1(-/-)Math1-cre:SmoM2 mice. Interestingly, tumors that developed in these mice displayed increased mitotic activity and decreased neuronal differentiation. Moreover, survival of these mice was significantly decreased. Similarly, low expression of BARHL1 in human medulloblastoma cases was associated with a less favorable prognosis for patients. These results suggest that the expression of Barhl1 decelerates tumor growth both in human and in murine medulloblastomas and should be further investigated with respect to potential implications for individualized therapeutic strategies.

[New aspects of MRI for diagnostics of large vessel vasculitis and primary angiitis of the central nervous system]. / Neue Aspekte der MRT-Bildgebung zur Diagnostik der Großgefäßvaskulitiden sowie der primären Angiitis des zentralen Nervensystems.

Vasculitis is a rare disease and clinical symptoms are often unspecific. Accurate and early diagnosis is mandatory in order to prevent complications, such as loss of vision or stroke. Imaging techniques can contribute to establishing a definite diagnosis and to evaluate disease activity and the extent of the disease in various vascular regions. Conventional imaging methods, such as computed tomography (CT) and magnetic resonance (MR) angiography, as well as digital subtraction angiography allow the vessel lumen but not the vessel wall to be depicted. However, vasculitis is a disease which primarily affects the vessel wall, therefore conventional imaging modalities often fail to make a definite diagnosis. Recently black-blood high resolution MR in vivo imaging has been used to visualize cervical and intracranial vasculitis. This review article presents imaging protocols for intracranial and cervical black-blood MR imaging and clinical cases with large vessel vasculitis and vasculitis of the central nervous system. Furthermore the current literature, examples of the most common differential diagnoses of cervical and cranial arteriopathy and the potential of other imaging modalities, such as PET/CT and ultrasound will be discussed.

Cerebellar stem cells act as medulloblastoma-initiating cells in a mouse model and a neural stem cell signature characterizes a subset of human medulloblastomas.

Cells with stem cell properties have been isolated from various areas of the postnatal mammalian brain, most recently from the postnatal mouse cerebellum. We show here that inactivation of the tumor suppressor genes Rb and p53 in these endogenous neural stem cells induced deregulated proliferation and resistance to apoptosis in vitro. Moreover, injection of these cells into mice formed medulloblastomas. Medulloblastomas are the most common malignant brain tumors of childhood, and despite recent advances in treatment they are associated with high morbidity and mortality. They are highly heterogeneous tumors characterized by a diverse genetic make-up and expression profile as well as variable prognosis. Here, we describe a novel ontogenetic pathway of medulloblastoma that significantly contributes to understanding their heterogeneity. Experimental medulloblastomas originating from neural stem cells preferentially expressed stem cell markers Nestin, Sox2 and Sox9, which were not expressed in medulloblastomas originating from granule-cell-restricted progenitors. Furthermore, the expression of these markers identified a subset of human medulloblastomas associated with a poorer clinical outcome.

Bcl-2 expression inversely correlates with tumour cell differentiation in medulloblastoma.

Medulloblastoma (MB) is a cerebellar primitive neuroectodermal tumour that occurs predominantly in childhood. It can be mainly divided into classical and desmoplastic tumours, but differential diagnosis is often difficult. Patients' prognosis is poor and neuropathological markers that reliably predict outcome are still missing. In a series of 104 MBs including 80 tumours of the classical and 24 tumours of the desmoplastic variant we studied the number of apoptotic figures and the expression of the proto-oncogene bcl-2, an anti-apoptotic protein known to affect tumour cell proliferation. We observed a strong correlation between the expression of bcl-2 with patients' age (P < 0.001) as well as with the desmoplastic subtype (P < 0.001). Here, protein expression was found to be restricted to internodular, less differentiated, highly proliferative areas. In classical MB, bcl-2 was detected only in 23% of cases and was highly inversely correlated with the expression of synaptophysin (P < 0.001) indicating that bcl-2 is predominantly expressed by undifferentiated classical MB. With regard to prognosis the expression of bcl-2 tended to correlate with poor outcome in classical MB but not in desmoplastic MB, although not to a statistically significant extension (P = 0.06). On the other hand, a high number of apoptotic figures in the tumour tissue was found to indicate poor prognosis independent of the histological subtype (P < 0.05).

Developmental expression of heterotrimeric G-proteins in the murine cerebellar cortex.

We utilized the murine cerebellum to analyze the expression of G-proteins during vertebrate neural differentiation. Combining reverse transcription-polymerase chain reaction and immunocytochemistry, we monitored the expression and cellular localization, within the nascent cerebellar cortex, of G-proteins subunits known to mediate signal transduction in the adult cerebellum. The mRNAs encoding subunits Galphaq, Galphao, Galphai-2, and Galphaz are expressed in the cerebellar anlage at least from embryonic day 14 onward, and relative levels of these mRNAs do not change appreciably from E14 to adulthood. Galphao, Galphaz, and Galphai-2 could be localized to granule cell neuroblasts and postmigratory, mature granule cells, but not to early postmitotic, premigratory, and migrating granule neurons. All of the Galpha subunits analyzed could also be localized to the cell somata of postmitotic Purkinje neurons, irrespective of age. In contrast, Purkinje cell dendrites stained for Galphao only up to postnatal day 8, dendritic immunoreactivity for Galphaz increased during dendritogenesis, and appreciable levels of Galphai-2 and Galphaq were seen in Purkinje cell dendrites only transiently during the 2nd and 3rd postnatal week. Of the G-beta and -gamma subunits analyzed (beta1, beta2, gamma2, gamma3, gamma5, and gamma7), only expression of gamma3 varied with development. It could be localized to Purkinje cell somata and dendrites in early postnatal, but not in adult animals. These changes in the cellular distribution and subcellular segregation of G-proteins are correlated to tangible aspects of cerebellar cortical histogenesis and suggest a role for G-protein-mediated signaling in their mechanistic implementation.

The intra-articular pressure of the shoulder: an experimental study on the role of the glenoid labrum in stabilizing the joint.

It was shown on human corpses that the glenohumeral joint may be compared to a piston surrounded by a valve. The glenoid labrum, then, should work like the valve block, sealing the joint from atmospheric pressure. In order to test this hypothesis, 18 human shoulder preparations were studied. The mean stabilizing force obtained by atmospheric pressure was 146 N. Additionally, 15 patients without any sign of instability and 17 patients with an anterior instability of the shoulder were tested under general anesthesia. In stable shoulder joints, traction at the arm caused negative intra-articular pressure that could be correlated to the amount of force exerted. In contrast, unstable shoulder joints with a tear of the glenoid labrum (Bankart lesion) did not exhibit this phenomenon. For unstable shoulder joints, the piston-and-valve model is no longer valid. This enlarges the current concept of shoulder joint stability in two ways: (a) the absence of negative intra-articular pressure disturbs joint mechanics and (b) altered pressure receptors might disturb motor coordination that dynamically protects the shoulder from dislocating forces.

[Significance of the glenoid labrum for stability of the glenohumeral joint. An experimental study]. / Die Bedeutung des Labrum glenoidale für die Stabilität des Glenohumeralgelenkes. Eine experimentelle Studie.

In human cadavers we were able to show that the glenohumeral joint is comparable to the model of a physical piston. The labrum glenoidale functions like a valve against atmospheric pressure. It is possible to characterize the behavior of intraarticular negative pressure by the equation (formula; see text). The calculations of the force F of atmospheric pressure tending to resist distraction of the joint surfaces leads to a 95% confidence interval from 6.9 to 22.9 kp. Under a general anesthetic, distraction of the healthy glenohumeral joint also produces negative intraarticular pressure in the area of the fossa glenoidalis in vivo. Joints with a labral tear (Bankart defect) and a chronic instability are not characterized by this phenomenon. A change in intraarticular pressure might stimulate intraarticular pressure receptors. This could be important in functioning as a neuromuscular protection reflex for the joint.