Brain Tumor 2008

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					                                       Brain Tumor 2008




Brain Tumor 2008




     Program and Abstracts
             (Orals and Posters)


        December 4 / 5, 2008

             Campus Berlin-Buch
 Max Delbrück Communications Center (MDC.C)
            Robert-Rössle-Str. 10
               D-13125 Berlin




           GRK 1258



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Brain Tumor 2008


                         Campus Berlin-Buch




A9 -    Pförtner                            C31.1- MDH
A8 -    Torhaus                             C31 - MDH - Max-Delbrück-Haus
A15 -   Charles River Deutschland GmbH      C84 - Hermann-von-Helmholtz-Haus
A14 -   Mensa                               C83 - MDC.C
A13 -   Infocenter, Gläsernes Labor         C81 - FMP
A10 -   Bibliothek                          C71 - Tier- und Laborgebäude
B64 -   epo GmbH                            D85 - Arnold-Graffi-Haus
B63 -   Tierhaus                            D82 - Karl-Lohmann-Haus
B61 -   Salvadore-Luria-Haus                D80 - Otto-Warburg-Haus
B55 -   OCVH - Oskar-und-Cécile-Vogt-Haus   D79 - Erwin-Negelein-Haus
B54 -   Hans-Gummel-Gästehaus               D72 - Haus 72
B46 -   Robert-Rössle-Klinik                D23 - Eckert & Ziegler AG
C27 -   Walter-Friedrich-Haus               D16 - Bebig GmbH
C87 -   Timoféeff-Ressovsky-Haus


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                                                        Brain Tumor 2008

                                    Table of Contents



Scientific Program                                          5

List of Oral Presentations selected from Abstracts          7

Abstracts of Oral Presentations                             9

List of Poster Presentations                               13

Abstracts of Poster Presentations                          17

Address List                                               27




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Brain Tumor 2008



                                 Welcome Address



Dear Colleagues,

We are pleased to announce that the Brain Tumor Meeting 2008 will take place at the Max
Delbrück Center for Molecular Medicine (MDC) in Berlin (Germany) on December 4 and 5,
2008.

In summer 2000 Berlin neuroscientists, neurosurgeons and neurologists focusing on brain
tumors initiated this scientific conference. It was repeated in 2001, 2004 and 2006 - throughout
this time the Brain Tumor Meeting gained national and international attention and attracted
many leading scientists working on gliomas and brain stem cells. The main focus of the
meeting is to provide a platform for an interdisciplinary scientific exchange especially between
scientists and clinicians.

Recent discoveries on the role of glioma stem cells, on the interaction of gliomas with their
microenvironment and on glioma cell death pathways have altered our understanding of
glioma biology. Internationally renowned speakers were invited to present their newest data
on these topics. Furthermore, abstracts for oral- and poster-presentations were invited and the
best presentations will be awarded a price.

We expect exciting scientific exchange and welcome you to join this meeting.

Scientific Program Committee




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                                                                                   Brain Tumor 2008



                             Scientific Program

                   Thursday,
                   Thursday, December 4, 2008



14.00 – 14.05   Welcome Address: Helmut Kettenmann

14.05 - 15.25   Session I
                Chair: Frank Heppner

14.05 – 14.45   Roger Abounader (Charlottesville, USA)
                The c-Met pathway in brain tumors

14.45 – 15.05   Ilker Eyupoglu (Erlangen, Germany)
                Molecular mechanisms of brain tumor induced cell death and edema formation

15.05 - 15.25   Gaetano Finocchiaro (Milano, Italy)
                Enhancer of ZESTA 2 is a relevant gene for cancer stem cells and glioma progression

15.25 – 16.00   Poster Session and Coffee Break

16.00 – 17.40   Session II
                Chair: Matthias Endres

16.00 – 16.40   Katrin Lamszus (Hamburg, Gemany)
                Phenotypic diversity of glioma stem cell lines: determinants, assessment and treatment of
                invasive glioma models

16.40 - 17.20   Maarten van Lohuizen (Amsterdam, Netherlands)
                Polycomb repressors controlling stem cell fate: Implications for cancer and development

17.20 – 17.40            Pollard
                Steven Pollard (Cambridge, UK)
                Glioma stem cell lines expanded in adherent culture have tumour-specific phenotypes and
                are suitable for chemical and genetic screens

17.40 – 18.00                    Poster
                Coffee Break and Poster Session

18.00 – 19.20   Session III
                Chair: Peter Vajkoczy

18.00 – 18.40   Peter B. Dirks (Toronto, Canada)
                      B.
                Tracking the cancer stem cells in human and mouse brain tumors

18.40 – 19.20   Stefan Momma (Frankfurt/M., Germany)
                The reaction of neural stem/progenitor cells to tumors in the adult human brain

19.30 – 20.00       Transfer            -Ruine
                                Virchow-R
                Bus Transfer to Virchow -Ruine

20.00           Reception in the Virchow Ruine / Charité




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Brain Tumor 2008


                         Friday, December 5, 2008
                         Friday,


9.00 - 10.00       Session IV
                   Chair: Achim Leutz

9.00 - 9.40             Martin-Villalba
                   Ana Martin-Villalba (Heidelberg, Germany)
                   Activation of survival signals by the death receptor CD95

9.40 - 10.00       Daniel Bexell (Lund, Sweden)
                   Multipotent mesenchymal stroma cells act as pericyte-like migratory vehicles
                   in experimental gliomas

10.00 – 10.30      Poster Session and Coffee Break

10.30 – 11.30      Session V
                   Chair: Jürgen Kiwit

10.30 – 11.10      Rolf Bjerkvig (Bergen, Norway)
                   Brain tumor stem cells, do they exist?

11.10 – 11.30      Gregor Auf (Talence, France)
                   IRE1 and glioma angiogenesis

11.30 – 12.30      Poster Session and Lunch
                                      Lunch

12.30 – 13.10      Session VI
                   Chair: Rainer Glass

12.30 – 12.50      Rossella Galli (Milano, Italy)
                   EGFR expression identifies functionally distinct subpopulations of tumor-initiating cells in
                   human glioblastoma multiforme

12.50 – 13.10                Tabatabai
                   Ghazaleh Tabatabai (Tübingen, Germany)
                   Experimental gliomas attract hematopoietic progenitor cells: a novel
                   approach towards a cellular therapy of malignant gliomas?

13.10 – 13.30      Poster Session and Coffee Break

13.30 – 14.50      Session VII
                   Chair: Frauke Zipp

13.30 – 14.10      Heidi Philipps (San Francisco, USA)
                   A hierarchy of self-renewing tumor-initiating cell types in glioblastoma

14.10 – 14.50      Sebastian Brandner (London, UK)
                   Neural stem cells out of control: the origin of brain tumours?

14.50              Awarding of Poster Prizes
                               Poster Prizes

15.00              Departure




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                                                                                            Brain Tumor 2008


         List of Oral Presentations selected from Abstracts


     Gregor
Auf, Grego
Université Bordeaux 1, INSERM U920, Talence, France, gregor.auf@charite.de
IRE1 AND GLIOMA ANGIOGENESIS
Gregor Auf, Sylvaine Guérit, Arnaud Jabouille, Raphaël Pineau, Marion Bouchecareilh, Sophie North, Alexandre
Favereaux, Marlène Maitre, Timo Gaiser, Andreas von Deimling, Eric Chevet, Andréas Bikfalvi, Michel Moenner

Bexell, Daniel
Lund Stem Cell Center, BMC B10, , Lund, Sweden, daniel.bexell@med.lu.se
MULTIPOTENT MESENCHYMAL STROMA CELLS ACT AS PERICYTE-LIKE MIGRATORY VEHICLES IN EXPERIMENTAL
GLIOMAS
Bexell, D.; Gunnarsson, S.; Tormin, A.; Darabi, A.; Gisselsson, D.; Roybon, L.; Scheding, S.; Bengzon, J.

Eyupoglu, Ilker
University of Erlangen-Nuremberg, Department of Neurosurgery, Erlangen, Gemany, ilker.eyupoglu@uk-erlangen.de
MOLECULAR MECHANISMS OF BRAIN TUMOR INDUCED CELL DEATH AND EDEMA FORMATION
Ilker Y. Eyüpoglu, Alexandra Heckel, Eric Hahnen, Tobias Engelhorn, Arnd Doerfler, Oliver Ganslandt, Christopher
Nimsky, Michael Buchfelder & Nicolai E. Savaskan

Finocchiaro, Gaetano
Fondazione IRCCS Istituto Neurologico Besta, Milano, Italy, elisabetta.griggio@istituto-besta.it
ENHANCER OF ZESTE 2 IS A RELEVANT GENE FOR CANCER STEM CELLS AND GLIOMA PROGRESSION
Orzan FN, Pellegatta S, Villa G, Caldera V, Eoli M, Farinotti M, Marras C, Pollo B, Schiffer D* and Finocchiaro G

Galli, Rossella
San Raffaele Scientific Institute, Stem Cell Research Institute, Milano, Italy, galli.rossella@hsr.it
EGFR EXPRESSION IDENTIFIES FUNCTIONALLY DISTINCT SUBPOPULATIONS OF TUMOR-INITIATING CELLS - IN
HUMAN GLIOBLASTOMA MULTIFORME
                                                .L.;
Galli, R.; Mazzoleni, S.; Politi, L.; Poliani, P De Palma, M.

Pollard, Steven
University of Cambridge, Wellcome Trust Centre for Stem Cell Research, Cambridge, UK, smp54@cam.ac.uk
GLIOMA STEM CELL LINES EXPANDED IN ADHERENT CULTURE HAVE TUMOUR-SPECIFIC PHENOTYPES AND
ARE SUITABLE FOR CHEMICAL AND GENETIC SCREENS.
Pollard, S.; Yoshikawa K.; Smith, A.; Dirks, P.

Tabatabai, Ghazaleh
University of Tübingen, Department of General Neurology, Tübingen, Germany, ghazaleh.tabatabai@uni-tuebingen.de
EXPERIMENTAL GLIOMAS ATTRACT HEMATOPOIETIC PROGENITOR CELLS: A NOVEL APPROACH TOWARDS A
CELLULAR THERAPY OF MALIGNANT GLIOMAS?
Tabatabai, G; Herrmann, C; Möhle, R; Weller; Wick, W




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                                Abstracts of Oral Presentations

C-MET PATHWAY IN BRAIN TUMORS                                         MULTIPOTENT MESENCHYMAL STROMA CELLS ACT AS
Roger Abounader                                                       PERICYTE-LIKE MIGRATORY VEHICLES IN EXPERIMENTAL
Neurology, University of Virginia, Charlottesville, USA               GLIOMAS
The receptor tyrosine kinase c-Met and its ligand hepatocyte          Daniel Bexell1,2,, Salina Gunnarsson1,2, Ariane Tormin2, Anna
growth factor (HGF) have emerged as key determinants of brain         Darabi1, David Gisselsson3, Laurent Roybon4, Stefan
tumor malignancy. We showed that c-Met and HGF are often              Scheding2, Johan Bengzon1,2
                                                                      1
                                                                        The Rausing Laboratory, Division of Neurosurgery, Dept. of
aberrantly expressed in gliomas and embryonal brain tumors.
                                                                      Clinical Sciences, 2Lund Strategic Research Center for Stem
We found that c-Met activation in brain tumors is associated with
                                                                      Cell Biology and Cell Therapy, 3Department of Clinical
poor clinical outcomes and is mostly caused by transcriptional
                                                                      Genetics, Lund University Hospital, 4Neuronal Survival Unit,
overexpression of receptor and ligand. We uncovered a role for
                                                                      Wallenberg Neuroscience Center, Department of
microRNA-34a downregulation in inducing c-Met overexpression
                                                                      Experimental Medical Science, Lund University
in brain tumors. c-Met activation in brain tumors enhanced
                                                                      Bone marrow-derived multipotent mesenchymal stroma cells
malignancy by inducing cell proliferation, inhibiting cell death,
                                                                      (MSCs) have emerged as cellular vectors for gene therapy of
inducing cell migration and cell invasion and promoting tumor
                                                                      solid cancers. We implanted enhanced green fluorescent protein-
angiogenesis. The oncogenic effects of HGF/c-Met were
                                                                      expressing MSCs directly into rat malignant gliomas to address
mediated by complex downstream signal transduction and
                                                                      their migratory capacity, phenotype and effects on tumor
transcription networks, the most prominent components of which
                                                                      neovascularization and animal survival. A single intratumoral
were Ras/MAPK, PI3K/Akt/mTOR and c-Myc. We demonstrated
                                                                      injection of MSCs infiltrated the majority of invasive glioma
that HGF and c-Met are potential targets for brain tumor therapy.
                                                                      extensions (72±14%) and a substantial fraction of distant tumor
We tested different approaches for inhibiting c-Met and HGF in
                                                                      microsatellites (32±6%). MSC migration was highly specific for
brain tumor models, including gene expression inhibition with
                                                                      tumor tissue. Grafted MSCs integrated into tumor vessel walls
ribozymes, neutralizing antibodies to HGF, and small molecule
                                                                      and expressed pericyte markers α-smooth muscle actin, neuron-
inhibitors of c-Met kinase. While these approaches were
                                                                      glia 2 and platelet-derived growth factor receptor−β but not
successful in inhibiting tumor growth in animal models, therapies
                                                                      endothelial cell markers. The pericyte marker expression profile
that target a single pathway such as HGF/c-Met will most likely
                                                                      and perivascular location of grafted MSCs indicate that these
benefit at best only a subset of human patients. HGF/c-Met
                                                                      cells act as pericytes within tumors. MSC grafting did not influence
pathway targeting will probably be most efficacious in combination
                                                                      tumor microvessel density or survival of tumor-bearing animals.
with other existing and emerging therapies. We demonstrated
                                                                      Intratumorally grafted pericyte-like MSCs might represent a
the value of combining anti-HGF/c-Met therapies with traditional
                                                                      particularly well suited vector system for delivering molecules to
cytotoxic therapies such as radiotherapy. We uncovered a
                                                                      affect tumor angiogenesis and for targeting cancer stem cells
connection between c-Met and EGFR pathways that provides a
                                                                      within the perivascular niche.
rationale for combining anti-c-Met with anti-EGFR therapies. We
also found that PTEN loss amplifies c-Met-induced glioblastoma
malignancy and that combining anti-HGF/c-Met approaches with
PTEN restoration or mTOR inhibition might have therapeutic
                                                                      BRAIN TUMOURS CAN ARISE FROM STEM/PROGENITOR
advantage.                                                            CELLS IN THE ADULT BRAIN THROUGH ACTIVATION OF
                                                                      ONCOGENIC PATHWAYS: EVIDENCE FOR PATHWAY-
                                                                      SPECIFIC TUMOUR PHENOTYPE
IRE1 AND GLIOMA ANGIOGENESIS                                          Sebastian Brandner1 Thomas S Jacques2, Heike Naumann1,
Gregor Auf, Sylvaine Guérit, Arnaud Jabouille, Raphaël Pineau,        Alexander Swales1, Arturo Alvarez-Buylla3
Marion Bouchecareilh, Sophie North, Alexandre Favereaux,              1
                                                                        Division of Neuropathology and Department of
Marlène Maitre, Timo Gaiser, Andreas von Deimling, Eric
Chevet, Andréas Bikfalvi, Michel Moenner                              Neurodegenerative Disease, UCL Institute of Neurology,
INSERM U920, Université Bordeaux 1, 43, Avenue des                    London WC1N 3BG, UK, 2Neural Development Unit, UCL-
Facultés, 33405 Talence, France                                       Institute of Child Health and Department of Histopathology,
Ischemia is associated to glioma development and locally induces      Great Ormond Street Hospital, London WC1N 1EH,
an adaptive response which confers to tumor cells an enhanced         UK,3Department of Neurosurgery and Developmental and
survival and a more agressive behaviour. Hypoxia or glucose           Stem Cell Biology Program, University of California, San
deprivation, characteristic features of tumor ischemia, activate      Francisco, San Francisco, CA 94143–0525, USA
complex intracellular signaling referred as to the Unfolded Protein   There is increasing evidence that brain tumours arise from
Response (UPR). IRE1, one of the UPR proximal sensors, is a           neoplastic transformation of neural stem cells which have the
key regulator of these effects. It has been previously shown          ability to self-renew and differentiate into neurons and glia,
that IRE1 inactivation correlated in vitro with the down-regulation   because (i) they contain multiple cell types, suggestive of an
of the pro-angiogenic factor VEGF-A in various tumor cell types       origin from a cell with multilineage potential; (ii) they often appear
including glioma-derived cells. To study the role of IRE1 in tumor    to arise from the ventricular zone, the location of NSC; (iii) they
development and angiogenesis, immunodeficient mice were               have in common the expression of certain stem cell markers; (iv)
subjected to intracerebral injection of human U87 glioma cells        they express genes that regulate proliferation and self-renewal
either expressing or not an IRE1 dominant negative transgene          of normal NSCs and mutations in genes that normally regulate
(U87 IRE1.DN). Animals bearing the U87 IRE1.DN-derived tumors         NSC proliferation are frequently found in IBTs and (v) forced
showed reduced tumor angiogenesis and growth as well as an            expression of oncogenes in neural stem and progenitors cells in
increased overall survival, as compared to the control mice.          mice produces tumours that are similar to primary human tumours.
Histopathological analysis of IRE1.DN tumors also revealed a          To determine how the type of tumour depends on the cell of
major phenotypic change suggestive of a mesenchymal drift. A          origin or the initial mutations, we used conditional knock-out mice
functional link between the blockade of IRE1 activity and tumor       to target the adult neural stem cell compartment by injecting
vascularization was further underlined by transcriptomic              Adenovirus expressing cre recombinase into the cerebral
analysis. The expression pattern of pro- and anti- angiogenic         ventricles. Also, we derived neurospheres following
genes in IRE1.DN expressing cells is consistent with the inhibition   recombination, expanded them in vitro and finally grafted them
of angiogenesis. In vivo validation of gene expression profiling      into recipient mice to test their growth, differentiation and the
was performed using the chicken chorioallantoic membrane              capacity to form tumours. Recombination of [Rb, p53] or of [Rb,
(CAM) assay as well as the mouse brain implantation model             p53 and PTEN] showed features of primitive neuroectodermal
coupled to laser capture microdissection analyses. Our results        tumours (PNETs), while inactivation of [PTEN; p53] resulted in
predict that IRE1 is a key regulator of glioma angiogenesis.          formait0n of diffuse gliomas. Stem cells derived from brains
                                                                      shortly after intraventricular recombination and after expansion



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Brain Tumor 2008


in vitro were grafted into recipient mice formed tumours, providing   EGFR EXPRESSION IDENTIFIES FUNCTIONALLY DISTINCT
further evidence that the initial recombination was in the stem       SUBPOPULATIONS OF TUMOR-INITIATING CELLS IN
cell compartment while in contrast, astrocytes did not give rise      HUMAN GLIOBLASTOMA MULTIFORME.
to tumours. In conclusion, we show that intrinsic brain tumours       Rossella Galli1, Stefania Mazzoleni1, Letterio Politi2, Pietro Luigi
                                                                      Poliani3, Michele De Palma4.
can indeed originate from CNS stem cells and that the initial         1
                                                                       Stem Cell Research Institute (SCRI) — San Raffaele
genetic event influences the phenotype of the tumour.
                                                                      Scientific Institute, Via Olgettina 58, Milan, Italy, 20132,
                                                                      2
                                                                       Neuroradiology Unit – San Raffaele Scientific Institute, Via
MOLECULAR MECHANISMS OF BRAIN TUMOR INDUCED                           Olgettina 60, Milan, Italy, 20132; 3Department of Pathology —
CELL DEATH AND EDEMA FORMATION                                        University of Brescia — Spedali Civili of Brescia, P.le Spedali
Ilker Y. Eyüpoglu1, Alexandra Heckel1, Eric Hahnen2, Tobias           Civili 1, Brescia, Italy, 25124; 4Angiogenesis and Tumor
Engelhorn3, Arnd Doerfler3, Oliver Ganslandt1, Christopher            Targeting Research Unit, and Telethon Institute for Gene
Nimsky1, Michael Buchfelder1 & Nicolai E. Savaskan4                   Therapy — San Raffaele Scientific Institute, via Olgettina 58,
1
 Department of Neurosurgery, University of Erlangen-                  Milan, Italy, 20132.
Nuremberg; 2Institute of Human Genetics, Institute of                 Glioblastoma multiforme (GBM), the most frequent malignant brain
Genetics and Center for Molecular Medicine Cologne                    tumor of adults, is characterized by enhanced tumor cell dispersal
(CMMC), University of Cologne; 3Department of                         into the brain parenchyma. This pathological trait, together with
Neuroradiology, University of Erlangen-Nuremberg;                     resistance to standard treatments, makes GBM prognosis highly
4
 Department of Neuromorphology, Brain Research Institute,             dismal. Since most of de novo GBMs over-express the epidermal
Swiss Federal Institute of Technology (ETH) & University              growth factor receptor (EGFR), pharmacological targeting by
Zurich.                                                               means of receptor tyrosine kinase inhibitors (TKI) has been
Malignant gliomas represent one of the most aggressive                proposed as potentially effective therapeutic approach in GBMs.
neoplasias with still poor prognosis. Aside from uncontrolled cell    Interestingly, EGFR has been shown to be expressed unevenly
proliferation, perifocal edema is a hallmark of these tumors and      in human GBMs; therefore, it might specifically label subsets of
contributes mainly to the morbidity and mortality. The mechanisms     GBM cells that could behave as tumor-initiating cells (TICs). To
by which these tumors cause neuronal degeneration and brain           test this hypothesis, we purified different GBM cell fractions
edema are still ill-defined, however it is assumed that vasogenic     from patient’s specimens by fluorescence activated cell sorting
effects accounts for the development of edemas. Here, we              (FACS), based on their relative expression of EGFR and of the
show that malignant gliomas induce perifocal edema by glutamate       putative CSC marker CD133. Immediately after cell sorting, all
secretion which also drives neuronal cell death. Screening for        GBM subpopulations were injected intracranially into nude mice.
candidate glutamate transporters in brain tumors revealed the         Remarkably, all these subpopulations were endowed with
antiporter xCT as the main source for glutamate secretion. RNA        tumorigenic potential, although to different extent. In particular,
interference mediated knock down or pharmacological inhibition        EGFR expression conferred to GBM TICs enhanced in vivo
of xCT leads to abrogated glutamate secretion in gliomas and          malignancy and, in some patients, a distinctive migratory and
reduces neurodegeneration. Moreover, perifocal edema, which           invasive behaviour. In conclusion, these observations allocate
imposes in T2-weighted MR scans as high intensity areas               the well-known expression of EGFR in GBMs to a highly
surrounding the bulk tumor mass, was significantly alleviated in      tumorigenic and malignant subset of TICs. At the same time, they
xCT knock down gliomas transplanted into rat brains and led to        open up new therapeutically relevant scenarios, as the presence
attenuated clinical deterioration and prolonged survival. Whether     of phenotypically and biologically distinct TIC subpopulations
neurodegeneration is a prerequisite for brain edema and to what       within the same tumour might affect clinical response to treatment.
extent glioma induced neuronal degeneration contributes to brain
edema is not defined currently. However, diminished glutamate
secretion clearly lessens neurodegeneration and alleviates the        PHENOTYPIC DIVERSITY OF GLIOMA STEM CELL LINES.
development of edema. This indicates that in addition to              DETERMINANTS, ASSESSMENT AND TREATMENT OF
uncontrolled proliferation xCT mediated glutamate excretion           INVASIVE GLIOMA MODELS
defines another clinically relevant malignancy feature of primary     Katrin Lamszus
brain tumors.                                                         Laboratory for Brain Tumor Biology, Dept. of Neurosurgery,
                                                                      University Medical Center Hamburg-Eppendorf, Germany
                                                                      With the prime intention to establish an invasive glioma in vivo
ENHANCER OF ZESTE 2 IS A RELEVANT GENE FOR CANCER                     model for therapeutic studiesm we generated 12 human
STEM CELLS AND GLIOMA PROGRESSION                                     glioblastoma stem cell lines (GS-lines), using conditions that select
Orzan FN, Pellegatta S, Villa G, Caldera V*, Eoli M, Farinotti M,     for the expansion of neural stem cells. Microarray and phenotypic
Marras C, Pollo B, Schiffer D* and Finocchiaro G                      analyses of 9 GS-lines showed that they formed two distinct
Fondazione IRCCS Istituto Neurologico Besta, Milano and               groups. Unsupervised analysis yielded two separate clusters,
*Fondazione Policlinico di Monza, Italy                               with 4 cell lines characterized by the expression of
Glioblastoma multiforme (GBM) and other tumors may contain a          neurodevelopmental genes. They showed a multipotent
fraction of cells endowed with stem cell properties (cancer stem      differentiation profile along neuronal, astroglial and
cells, CSC), essential for tumor perpetuation. We used the            oligodendroglial lineages, grew spherically in vitro, expressed
neurosphere (NS) assay to screen for genes relevant to the            CD133, and formed highly invasive tumors in vivo. The other 5
CSC phenotype. GBM-NS were obtained in 50% of the GBM                 cell lines shared expression signatures enriched for extracellular
examined: GBM from which NS could be derived were associated          matrix-related genes, had restricted differentiation capacity,
to significantly shorter overall survival (p=0.014), suggesting       contained no or fewer CD133+ cells, grew adherent in vitro, and
that NS formation is depending on the biological features of the      displayed reduced tumorigenicity in vivo. These findings
tumor. Using a combination of DNA microarray and real time PCR        demonstrate that glioblastoma cell lines with a full stem-like
we found that GBM-NS express high levels of a component of            phenotype express neurodevelopmental genes as a distinctive
the Polycomb Repressive Complex 2 (PRC2), Enhancer of Zeste           feature. In parallel, we pursued another strategy to produce an
2 (EZH2). EZH2, a key player in stem cell maintenance, was            invasive glioblastoma model. Tumor tissue was briefly cultured
significantly more expressed in GBM than in low-grade gliomas         as spheroids, which were injected into the brains of nude mice.
as shown by real time PCR (p<0.001) and by                            Highly invasive tumors were obtained, histologically
immunohistochemistry of different gliomas. Targeting EZH2 by          indistinguishable from those generated by GS-lines with a full
siRNA favoured NS differentiation and decreased expression of         stem-like phenotype. Using this model, we evaluated the effects
the CSC marker CD133. Treatment of GBM-NS with histone                of antibodies against EGFR (cetuximab, infused locally) and
deacetylase inhibitor SAHA (2 microM) decreased EZH2                  VEGFR-2 (DC101, intraperitoneally). Three of 7 cases responded
expression to 36% and CD133 expression to 4.4%. These results         to cetuximab with 70-80% tumor growth inhibition, but 4 were
identify EZH2 as a relevant gene for CSC biology and encourage        non-responsive. All responsive cases but no non-responsive
their therapeutic targeting by HDAC inhibitors.



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                                                                                                                Brain Tumor 2008


ones were derived from glioblastomas exhibiting EGFR gene               A HIERARCHY OF SELF-RENEWING TUMOR-INITIATING
amplification and expression of EGFRvIII, suggesting that               CELL TYPES IN GLIOBLASTOMA
responsiveness to cetuximab depends on EGFR status. In                  Ruihuan Chen1, Merry C. Nishimura1, Stephanie M. Bumbaca1,
contrast, none of 4 xenograft cases treated with DC101                  Samir Kharbanda1, William F. Forrest2, Ian M. Kasman1, Joan M.
                                                                        Greve1, Robert H. Soriano3, Laurie L. Gilmour4, Zora
responded to treatment, and tumors lacked obvious angiogenic            Modrusan3 , Serban Nacu5, Katrin Lamszus6, Manfred
activity.                                                               Westphal6, Susanne Heim7 , C. David James8, Scott R.
                                                                        VandenBerg8, Cynthia J. Cowdrey8, Michael Prados8, & Heidi S.
                                                                        Phillips1,8
                                                                        Departments of Tumor Biology and Angiogenesis1,
ACTIVATION OF SURVIVAL SIGNALS BY THE DEATH                             Biostatistics2, Molecular Biology3, Immunology4 , and
RECEPTOR CD95                                                           Bioinformatics5, Genentech Inc.; University of Hamburg
Ana Martin-Villalba                                                     Eppendorf6; DefiniensAG7, Munich Germany and the Brain
DKFZ, INF 581, 69120 Heidelberg, Germany                                Tumor Research Center8, University of California San
Glioblastoma multiforme typically have diffuse, infiltrative growth     Francisco.
patterns which hinder the achievement of complete surgical              The concept that human malignancies are relentlessly fueled by
resection. Current treatment modalities aim at inducing apoptosis       identifiable subpopulations of tumor cells with unlimited capacity
of remaining tumor cells in part by inducing expression of the          for self-renewal has important therapeutic implications, yet
death system CD95/CD95L. Overcoming resistance of tumor cells           remains a hotly debated issue. In glioblastoma mulitforme (GBM),
to CD95-induced apoptosis has been in the limelight of cancer           the most common adult primary brain tumor, CD133 is a marker of
research. We, however, found that tumor cells take advantage            cells that demonstrate self-renewal and tumor initiation in
of CD95 to increase their infiltration capacity. Further, tumor cells   experimental conditions, however, instances of CD133- cells
even induce expression of the ligand for CD95 in the surrounding        exhibiting similar properties have also been reported. To examine
tissue. Thus, contrary to expectations, inhibition and not induction    heterogeneity of tumor cell competencies in GBM, we examine a
of the death system CD95/CD95L should be considered as front-           series of patient tumors and tumor-derived neurosphere lines.
line therapy for GBM.                                                   Here we show that individual GBM cases contain a hierarchy of
                                                                        CD133- and CD133+ self-renewing, tumor-initiating cell types
                                                                        and provide evidence that these cell types constitute a lineage
                                                                        hierarchy. Clonal analysis reveals three self-renewing cell types,
THE REACTION OF NEURAL STEM/PROGENITOR CELLS TO                         defined on the basis of CD133 expression in the parental cell
TUMORS IN THE ADULT HUMAN BRAIN                                         and its progeny. Surprisingly, all three cell types can not only
Stefan Momma1*, Jadranka Macas1, Christian Scharenberg2,                undergo long-term self-renewal in culture but also initiate serially
Marius Dohse3, Christian Nern1, Josefine von Randow1, Kea               transplantable tumors in nude mice. Grafts of lines derived from
Franz4, Volker Seifert4, Michael Synowitz5, Rainer Glass6, Karl
H. Plate1, Cornelia Brendel7                                            each of the self-renewing cell types display differences in growth
Institute of Neurology (Edinger Institute)1, Department of              kinetics, histological features, and molecular signatures that are
Neurosurgery4, University of Frankfurt Medical School,                  consistent with the hypothesis that the cell types identified
Frankfurt am Main, Germany; 2Karolinska Institute,                      represent stages along a differentiation continuum. Importantly,
Department of Hematology, Stockholm, Sweden; 3National                  our findings reveal a population of CD133- cells that appear to
Institutes of Health, Bethesda, USA; 5Department of                     give rise to previously identified CD133+ stem-like cells in GBM.
Neurosurgery, Charité-Universitätsmedizin, Berlin; 6Max                 These findings confirm that CD133 is a marker that defines
Delbrück Center, Cellular Neuroscience, Berlin; 7Department             functionally distinct populations of GBM tumor cells, but reveal
of Hematology, University Hospital Marburg, Germany.                    that growth of GBMs may be driven by a lineage of self-renewing
The adult human brain retains the capacity to generate new              tumor cells exhibiting a spectrum of markers and competencies.
neurons in the hippocampal formation and neuronal progenitor            Our findings suggest that optimal treatment of GBM is likely to
cells (NPCs) in the forebrain even though the cytoarchitecture of       require targeting self-renewing tumor cell populations that
the subventricular zone of the lateral ventricles differs               encompass a range of phenotypic states.
significantly from that of mouse brains. Despite a decreasing
number of NPCs migrating along the olfactory tracts with age,
the adult human brain retains the capacity to react to ischemic         GLIOMA STEM CELL LINES EXPANDED IN ADHERENT
injury by an increase in proliferation and generation of NPCs           CULTURE HAVE TUMOUR-SPECIFIC PHENOTYPES AND ARE
close to the lateral walls of the lateral ventricles. If and to what    SUITABLE FOR CHEMICAL AND GENETIC SCREENS.
                                                                        Steven Pollard1, Koichi Yoshikawa2, Ian Clarke2, Davide
extend this injury reaction applies to tumors as well is currently      Danovi1, Stefan Stricker1, Roslin Russell1, Austin Smith1 and
unknown. We analyzed postmortem tissue from normal and                  Peter Dirks2
pathological human brain tissue as well as intraoperative               1
                                                                          Wellcome Trust Centre for Stem Cell Research and
specimens from neurosurgical tumor resections to study the              Department of Biochemistry, University of Cambridge, Tennis
cellular response to tumors in the forebrain. We observed that          Court Road, Cambridge, CB2 1QR, United Kingdom, 2 The
the generation of NPCs increased significantly in brains of             Hospital for Sick Children, University of Toronto, Toronto,
individuals that suffered from tumors that were located close to        Canada
the lateral ventricular walls. Additionally, NPCs accumulated at        Human brain tumours appear to have a hierarchical cellular
the tumor border. Further characterization of these NPCs                organisation, suggestive of a stem cell foundation. Here we
suggests that they are not derived from the tumor itself. We            demonstrate routine derivation of adherent cell lines from
identified a subpopulation of cells by flow cytometry carrying          malignant glioma that display stem cell properties and initiate high
markers of undifferentiated cells. This population was enriched         grade gliomas following xenotransplantation. Significantly, GNS
at the lateral walls of the lateral ventricles compared to cortex       cell lines from different tumours exhibit divergent gene expression
and cerebellum. Using cultivated adult human neural stem cells,         signatures and differentiation behaviour that correlate with
we investigated the interaction between neural stem/progenitor          specific neural progenitor subtypes. The diversity of gliomas
cells and tumor cells in vitro. In conclusion, the necessary            may therefore reflect distinct cancer stem cell phenotypes.
conditions that endogenous stem/progenitor cells in the adult           Adherent GNS cell lines offer significant experimental advantages
human brain react to tumors are satisfied. This leaves the option       compared to ‘sphere’ based cultures, and provide a simplified
for a possible strategy in employing neural stem/progenitor cells       model enabling refined studies of cancer stem cell behaviour. A
for therapeutical use.                                                  proof-of-principle live cell imaging-based chemical screen (450
                                                                        FDA-approved drugs) identifies both differential sensitivities of
                                                                        GNS cells and a common susceptibility to perturbation of serotonin
                                                                        signalling.




                                                                                                                                        11
Brain Tumor 2008


EXPERIMENTAL GLIOMAS ATTRACT HEMATOPOIETIC
PROGENITOR CELLS: A NOVEL APPROACH TOWARDS A
CELLULAR THERAPY OF MALIGNANT GLIOMAS?
G. Tabatabai,1 C. Herrmann, 1 R. Möhle,2 M. Weller1,3 & W. Wick1,4
1
 Department of General Neurology and 2Department of
Internal Medicine II, University of Tübingen; 3Department of
Neurology, University of Zurich; 4Clinical Cooperation Unit
Neurooncology, German Cancer Research Center,
Heidelberg
Stem cells are considered as attractive candidates for a cell-
based gene delivery to cancers. We investigated the potential of
CD34+ hematopoietic stem/progenitor cells (HPC) as autologous
cellular vehicles in the treatment of malignant gliomas. Experimental
gliomas attracted HPC in vivo by a transforming growth factor
(TGF)- β-regulated CXC chemokine ligand (CXCL) 12-dependent
pathway that was further enhanced by metalloproteinase-9-
mediated stem cell factor cleavage. Supernatants of irradiated
or hypoxic LNT-229 glioma cells augmented HPC migration in
vitro. CXCL12 promoter activity was enhanced in glioma cells
after irradiation or after exposure to hypoxia. The irradiation-
and hypoxia-induced release of CXCL12 was mediated by
hypoxia inducible factor-1α. Cerebral irradiation after orthotopic
glioma implantation enhanced the glioma tropism of HPC in vivo.
Further, we studied the role of adhesive interactions. Exposure
of human cerebral endothelial cells to supernatants of glioma
cells induced CD62E expression. Transendothelial migration
assays showed enhanced HPC migration after CD62E induction,
which was impaired by neutralizing CD62E antibodies. Glioma
supernatant-mediated induction of CD62E on endothelial cells
required TGF-β signaling in glioma cells as well as VEGF receptor-
2 signaling in endothelial cells. Taken together, HPC might be
promising vectors in the treatment of gliomas. Given the essential
roles of TGF-β and VEGF signaling, a potential HPC-based therapy
might not be compatible in combination with TGF-β-antagonistic
or VEGF-A-neutralizing approaches. Instead, HPC-based therapy
might be compatible with irradiation and chemotherapy.




12
                                                                                                                 Brain Tumor 2008



                                      List of Poster Presentations


                 Valerie
1. Albrecht, Valerie
Neurosurgical Clinic Grosshadern, Ludwig-Maximilians-University, 81377 München, Germany, valerie.albrecht@med.uni-muenchen.de
ADULT
ADULT HUMAN MESENCHYMAL STEM CELLS AND GLIOMA CELLS FORM A STRUCTURAL AS WELL AS A
FUNCTIONAL SYNCYTIUM IN VITRO
Albrecht, V.; Korte, B.; Buchner, A.; Riesenberg, R.; Mysliwietz, J.; Paron, I.; Tonn, J.-C.; Goldbrunner, R.; Schichor, Ch.

2. Barciszewska, Anna-Maria
Chair and Clinic of Neurosurgery and Neurotraumatology, Karol Marcinkowski University of Medical Sciences, 60-355 Poznan, Poland,
abarcisz@man.poznan.pl
                    METHYLATION ANALYSIS
GENOMIC DNA METHYLA TION ANALYSIS IN HUMAN BRAIN GLIOMAS
Barciszewska, A-M; Nowak, S.; Gawronska, I.; Barciszewska, M.

     Bernreuther,
3. Bernreuther, Christian
Institute of Neuropathology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany c.bernreuther@uke.uni-
hamburg.de
                                 -RECEPTOR
                            EGF-RECEPT
EXPRESSION OF THE EGF -RECEPTOR AND NEURONAL MARKERS IN GLIAL NEOPLASMS : A TISSUE              NEOPLASMS:
MICROARRA ARRAY
MICRO ARRAY        APPROA
                   APPRO A CH
Bernreuther, C.; Kamradt, T.; Kolevatova, L.; Kellner, A.; Matschke, J.; Simon, R.; Glatzel, M.

     Birkenmeier,
4. Birkenmeier, Gerd
Institute of Biochemistry, University of Leipzig, 04103 Leipzig, Germany, Gerd.Birkenmeier@medizin.uni-leipzig.de
TUMOR-                      ACTIONS                               -2-MACROGL
                                                           ALPHA-2-MA CROGLOBULIN AT
TUMOR - SUPPRESSIVE A CTIONS OF HUMAN ALPHA -2-MA CROGLOBULIN AT ASTROCYTIC TUMOR CELLS
Lindner, I.; Buchold, M.; Bigl, M.; Hutschenreuther, A.; Thal, D.

5. Castriconi, Roberta
16155 Genova, Italy, Roberta.Castriconi@unige.it,
NATURAL                     MEDIATED
NA TURAL KILLER CELL MEDIATED RECOGNITION OF HUMAN BRAIN TUMORS
Castriconi, R.; Dondero, A.; Bellora, F.; Daga, A.; Raso, A.; Moretta, L.; Moretta, A.; Bottino, C.

   Cheray,
6. Cheray, Mathilde
Laboratory of Neuroimmunologie, Faculté de Médecine, Université de Limoges, 87025 Limoges; France, mathilde.cheray@etu.unilim.fr
SOR TING AND RELATIONSHIP BETWEEN N- AND S -TYPE CELLS IN HUMAN NEUROBLASTOMA CELL LINE B Y
SORTING           RELATIONSHIP                                                          NEUROBLASTOMA                     BY
SEDIMENTA                               CTIONATION
                           FLOW FRACTIONA                                CELLULAR
SEDIMENTATION FIELD FL OW FRA CTIONATION (SDFFF). A CELL ULAR MODEL FOR CHEMOTHERAPY                 CHEMOTHERAPY
STUDIES.
                                         .;                          .J.P
Cheray, M.; Bégaud-Grimaud, G.; Lazcoz, P Castresana, J.S.; Cardot, P .; Jauberteau, M.O.; Lalloué, F.; Battu, S.

7. Cholewa, Jadwiga
Developmental Neurobiology, MDC, 13125 Berlin, Germany, jadwiga.cholewa@mdc-berlin.de
MODULATION                                       PROPERTIES BY             XSACKIEVIRUS ADENOVIRUS RECEPTOR
                                                                        COXSA
MODULATION OF NEURONAL MEMBRANE PROPER TIES B Y THE CO XSA CKIEVIRUS - ADENO VIRUS RECEPTOR
Cholewa J., Jüttner R. and F. G.Rathjen

8. Ciechomska, Iwona
Department of Cell Biology, Institute of Experimental Biology, 02-093 Warsaw, Poland, jcjech@nencki.gov.pl
                           UTOPHA
                         AUTOPHAG TO                                 DEATH
CONTRIBUTION OF A UTOPHA G Y TO DRUG -INDUCED DEA TH OF MALIGNANT GLIOMA CELLS
Ciechomska, I.; Kaminska, B.

9. Clément, Virginie
Centre Romand de Neurochirurgie, Geneva University Hospitals and University of Geneva, 1211 Geneva; Switzerland,
virginie.clement@medecine.unige.ch
             MORPHOLOGOGY           AUTOFLUORESCENCE
                                      UTOFL                                   SELF-RENEWING
SPECIFIC MORPHOL OG Y AND A UTOFL UORESCENCE IDENTIFY SELF-RENEWING AND TUMOR -INITIATING                        -INITIATING
                                                                                                       TUMOR-INITIA
GLIOMA CELLS
Clement, V.

10. Colin, Carole
INSERM U911 - CRO2, Université de la Méditerranée - Faculté de Médecine Timone, 13005 Marseille; France, carole.colin@univmed.fr
PILOCYTIC ASTROCYTOMA                                     PATHWA
PIL OCYTIC ASTROCYTOMA OF THE OPTIC PATHW A Y: A TUMOR DERIVING FROM RADIAL GLIA CELLS WITH
                       SIGNATURE
SPECIFIC GENE SIGNA TURE
Tchoghandjian, A.; Fernandez, C.; Colin, C.; El Ayachi, I.; Voutsinos-Porche, B.; Fina, F.; Scavarda, D.; Piercecchi-Marti, M.D.; Intagliata,
D.; Ouafik, L‘H.; Fraslon-Vanhulle, C.; Figarella-Branger, D.

11. Corno, Daniela
Stem Cell Research Institute, San Raffaele Scientific Institute, 20132 Milan, Italy, corno.daniela@hsr.it
IDENTIFICATION
IDENTIFICATION OF CANCER STEM CELLS IN A MOUSE MODEL OF MEDULL OBLASTOMA AND MOLECULAR             OBLASTOMA
                                                                                           MEDULLOBLAST
COMPARISON
COMPARISON WITH NORMAL NEURAL STEM CELLS .                     CELLS.
Corno, D.; Cipelletti, B.; Pala, M.; Politi, L.; Bulfone, A.; Galli, R.

12. Czabanka, Marcus
Dept. of Neurosurgery, Universitätsmedizin Charite, 13353 Berlin; Germany, marcus.czabanka@charite.de
                                                 HEMODY
EFFECTS OF SUNITINIB ON TUMOR HEMOD YANAMICS AND DELIVER Y OF CHEMOTHERAPY DELIVERY        CHEMOTHERAPY
Czabanka M.; Vinci M.; Heppner F  .;Ullrich A.;Vajkoczy P.




                                                                                                                                         13
Brain Tumor 2008



13. Dondero, Alessandra
Department of Experimental Medicine, University of Genova, 16132 Genova, Italy, alessandra.dondero@unige.it
BOTH                                                                 OBLASTOMA
                                                 NEGATIVE MEDULLOBLAST
BOTH CD133 POSITIVE AND CD133 NEGATIVE MEDULL OBLAST OMA CELL LINES EXPRESS LIGANDS FOR
                     RECEPTORS                                              -MEDIATED CYTO
                                                                  TO NK-MEDIA
TRIGGERING NK RECEPT ORS AND ARE SUSCEPTIBLE TO NK -MEDIATED CYTOTO XICITY
                                     .;                    .;
Dondero, A.; Castriconi, R.; Negri, F Bellora, F.; Nozza, P Carnemolla, B.; Raso, A.; Moretta, L.; Moretta, A.; Bottino, C.

14. Fuchs, Heiko
Center for Anatomy, Institute for Cell biology and Neurobiology, 10115 Berlin, Germany, heiko.fuchs@charite.de
   FEEDBACK LOOP                                            LET-7                   -LET-7 MATURA
                                                                                 PRE-LET
A FEEDBA CK L OOP COMPRISING LIN-28 AND LET-7 CONTROLS PRE -LET-7 MA TURATION DURING NEURAL      TURATION
STEM- CELL COMMITMENT.
               COMMITMENT.
Fuchs H.,Rybak A., Smirnova L., Nitsch R., Wulczyn FG.

15. Gabrusiewicz, Konrad
Department of Cell Biology, The Nencki Institute of Experimental Biology, 02-093 Warsaw, Poland, k.gabrusiewicz@nencki.gov.pl
BLOCKING                                  TRATION
                                    INFILTRA                   CTIVA
                                                             ACTIV
BL OCKING OF MICROGLIAL INFILTRATION AND A CTIV A TION IN TRANSPLANTED GLIOMAS WITH
SYSTEMICALLY APPLIED CY CL OSPORIN A SIGNIFICANTLY REDUCES TUMOR GROWTH
SYSTEMICALLY                     CLOSPORIN
                              CYCL                   SIGNIFICANTLY
Gabrusiewicz, K.; Porycka, M.; Frankowska, M.; Kaminska, B.

16. Gaunitz, FrankFrank
Institut of Biochemistry, University of Leipzig, 04103 Leipzig, Germany, fgau@medizin.uni-leipzig.de
CARNOSINE INHIBITS GROWTH OF CELLS DERIVED FROM HUMAN GLIOBLASTOMA MULTIFORME AND IN    GLIOBLASTOMA MULTIFORME
AN ANIMAL MODEL OF HER2/NEU B Y INTERFERENCE WITH ANAEROBIC GLY COLYSIS
                                                 BY                                            GLY COLYSIS
Gaunitz, F.; Renner, C.; Seyffarth, A.; Asperger, A.; Zemitzsch, N.; Gebhardt, R.; Meixensberger, J.

17. Glass, Rainer
Cellular Neurosciences, MDC, 13125 Berlin, Germany, rainer.glass@mdc-berlin.de
                                                                         TTENUA
                                                                       ATTENU
BMP7 RELEASE FROM ENDOGENOUS NEURAL PRECURSORS ATTENU A TES TUMORIGENICITY OF GLIOMA
INITIATING
INITIATING CELLS
Chirasani SR1, Sternjak A2, Wend P3, Momma S4, Herold-Mende C5, Besser D3, Synowitz M6 & Kettenmann H1 Glass R1,

18. Hattermann, Kirsten
Anatomie, Universität Kiel, 24098 Kiel, Germany, k.hattermann@anat.uni-kiel.de
                                                             RECEPTOR CXCR7
EXPRESSION AND FUNCTION OF THE CHEMOKINE RECEPTOR CX CR7 IN HUMAN GLIOMAS
Hattermann, K.; Held-Feindt, J.; Lucius, R.; Mentlein, R.

19. Hoene, Victoria
Neonatology, Charité Universitätsmedizin Berlin, CVK, 13353 Berlin, Germany, victoria.hoene@charite.de
GATA -2 AND -3 ARE HIGHLY EXPRESSED IN NEUROBLASTOMA OF FA V ORABLE SUBTYPE
GAT                         HIGHLY                       NEUROBLASTOMA            FA
Hoene, V.; Fischer, M.; Ivanova, A.; Berthold, F.; Dame, C.

20. Holland, Heidrun
University of Leipzig, Translational Centre for Regenerative Medicine, 04103 Leipzig, Germany, Heidrun.Holland@medizin.uni-
leipzig.de
                                        CYTOGENETIC CHARACTERIZA    CTERIZATION
COMPREHESIVE (MOLECULAR) CYTOGENETIC CHARA CTERIZA TION OF RARE INTRA CRANIAL TUMORS            INTRACRANIAL
Holland, H.; Krupp, W.; Koschny, R.; Livrea, M.; Schober, R.; Meixensberger, J.; Ahnert, P.

21. Kahlert, Ulf
Stereotactic Neurosurgery, University Hospital Freiburg, 79106 Freiburg, Germany, ulf-dietrich.kahlert@uniklinik-freiburg.de
DEFINING IN VITRO CONDITIONS FOR ENRICHMENT OF STEM-LIKE CELL POPULATION IN HUMAN BRAIN       POPULATION
TUMOR CULTURES
            CULTURES
Kahlert, U.; Maciaczyk, J.; Nikkhah, G.

22. Kirsch, Matthias
Neurochirurgie, Carl Gustav Carus Universitätsklinikum Dresden, 01307 Dresden, Germany, matthias.kirsch@uniklinikum-dresden.de
                                    IMAGING TOW   OWARDS
INFRARED SPECTROSCOPIC IMA GING : TOW ARDS A METHOD FOR LABEL FREE CHARA CTERIZATION OF           CHARACTERIZA
                                                                                                          CTERIZATION
NEURAL STEM CELLS
Matthias Kirsch, Saskia Küchler, Andreas Hermann, Edmund Koch, Reiner Salzer, Gabriele Schackert, Gerald Steiner

23. Kruttwig, Klaus
in-vivo-NMR-Laboratory, Max-Planck-Institute for neurological research, 50931 Köln, Germany, kruttwig@nf.mpg.de
DEVELOPMENT                                                    LONGITUDINAL OBSERVA
DEVEL OPMENT OF A 3D IN VITRO MODEL FOR L ONGITUDINAL OBSERV ATION OF CELL A GGREGATES                       GGREGATES
                                                                                                           AGGREGA
COMBINING HIGH FIELD MAGNETIC RESONANCE IMAGING (MRI) WITH OPTICAL TECHNIQUE
SK. Kruttwig; C. Brueggemann; E. Kaijzel; S. Vorhagen; T. Hilger; C. Löwik; M. Hoehn

24. Lipko, Maciej
Cell Biology, NENCKI Institute of Experimental Biology, 02-093 Warsaw, Poland, mlipko@nencki.gov.pl
GLOBAL GENE EXPRESSION PROFILING REVEALS DISTINCT GENOMIC RESPONSES IN PRIMARY MICROGLIAL
    LT
C U LT U R E S
Lipko, M.; Dabrowski, M.; Kaminska, B.

25. Markovic, Darko
Neurosurgery, Helios Klinikum, 13125 Berlin, Germany, darko.markovic@helios-kliniken.de
                              EXPLOIT
GLIOMA INDUCE AND EXPL OIT MICROGLIAL MT1-MMP EXPRESSION FOR TUMOR EXPANSION                            EXPANSION
Markovic, DS; Chirasani, S; Synowitz, M; Sliwa, M; van Rooijen, N; Holmbeck, K; Kiwit, J; Kaminska, B; Glass, R; Kettenmann, H




14
                                                                                                             Brain Tumor 2008



     Parmaksiz,
26. P armaksiz, Güliz
Department of Neurosurgery, Charite Universitaetsmedizin, 10115 Berlin, Germany, gueliz.parmaksiz@charite.de
IN VIVO BIODISTRIBUTION AND MICROVASCULAR BINDING OF A HIGH-AFFINITY MONOCLONAL
ANTIBODY FRAGMENT F8-SIP AGAINST THE EXTRA-DOMAIN A OF FIBRONECTIN
Parmaksiz, G.; Czabanka, M.; Neri, D.; Vajkoczy, P.

     Porycka,
27. P orycka, Malgorzata
Department of Cell Biology, The Nencki Institute of Experimental Biology, 02-093 Warsaw, Poland, m.porycka@nencki.gov.pl
                                                            CROPHAGES TO
                                                         MACROPHA
CONTRIBUTION OF PERIPHERAL AND BRAIN MA CROPHA GES TO GLIOMA PA THOGENESIS AND THE EFFECT   PA
OF SYSTEMIC CY CL OSPORIN A TREATMENT
                 CYCLCLOSPORIN          TREATMENT
Porycka, M.; Gabrusiewicz, K.; Kwiatkowska, E.; Kowalczyk, D.; Kaminska, B.

28. Rybak, Agnieszka
Center for Anatomy, Institute for Cell biology and Neurobiology, 10115 Berlin, Germany, agnieszkarybak@yahoo.com
EXPRESSION AND FUNCTION OF THE LET7 MICRORNA DURING MOUSE CNS DEVELOPMENT
Rybak, A.; Fuchs, H.; Smirnova, L.; Nitsch R.; Wulczyn G.

29. Seiz, Marcel
Universitätsmedizin Mannheim, Neurochirurgische Universitätsklinik, 68167 Mannheim, Germany, marcel.seiz@nch.ma.uni-
heidelberg.de
DIFFERENTIATION                                     TRANSFORMATION
DIFFERENTIA TION BETWEEN MALIGNANT TRANSFORMATION AND TUMOUR RECURRENCE B Y 68GA -                               BY 68GA-
BOMBESIN AND 18F -FDG -PET IN PATIENTS WITH L OW GRADE GLIOMAS
                     18F-FDG              PA                 LOW
Seiz, M.; Dimitrakopoulou-Strauss A,; Schubert, G.A.; Brockmann, C.; Strauss, L.G.; Eisenhut, M.; Tüttenberg, J.

      Tchoghandjian,
30. Tchoghandjian, Aurélie
INSERM U911 - CRO2, Université de la Méditerranée - Faculté de Médecine Timone, 13005 Marseille, France,
aurelie.tchoghandjian@univmed.fr
                                   GLIOBLASTOMA HAVE
A2B5 CELLS FROM HUMAN GLIOBLASTOMA HA VE CANCER STEM CELL PROPER TIES                 PROPERTIES
Aurélie Tchoghandjian1*, Nathalie Baeza1*, Carole Colin1, Myriam Cayre2, Philippe Metellus3, Christophe Beclin2, L’Houcine Ouafik1
and Dominique Figarella-Branger1, 4

      yburczy,
     Tyburczy
31. Tyburczy, Magdalena
Department of Cell Biology, Nencki Institute of Experimental Biology, 02-093 Warsaw, Poland, m.tyburczy@nencki.gov.pl
IDENTIFICATION OF GENES REGULATED B Y MT OR IN TUBEROUS SCLEROSIS AND MALIGNANT GLIOMAS
IDENTIFICATION                       REGULATED BY MTOR
                                                    .;
Tyburczy M.E.; Kotulska K.; Jozwiak S.; Pokarowski P Mieczkowski J.; Kaminska B.

32. vom Berg, Johannes
Institute of Experimental Immunology, Department of Pathology, University Hospital of Zurich, 8057 Zurich, Switzerland,
johannes.vomberg@neuroimm.uzh.ch
                  CYTOKINE                   MEDIATING INTRACRANIAL
DISSECTING CYTOKINE NETWORKS MEDIA TING INTRA CRANIAL TUMOR CONTROL
vom Berg, J.; Haller, S.; Becher, B.

    Watson,
33. W atson, Deborah
Neurosurgery, University of Pennsylvania, 19146 Philadelphia, PA, USA, djw3@mail.med.upenn.edu
                     PA                                  MIGRATORY
BRDU LABELING P ATTERN OF THE ROSTRAL MIGRATOR Y STREAM IN NORMAL CANINE AND FELINE BRAINS
Watson, D.; Malik, S.; Vite, C.; Haskins, M.; Van Winkle, T.

     Wisniewski, Pawel
34. W isniewski, Pawel
Department of Cell Biology, The Nencki Institute of Experimental Biology, 02-093 Warsaw, Poland, p.wisniewski@nencki.gov.pl
FAS SIGNALING REGULATES INV ASIVENESS OF GLIOMA CELLS AND MMP-2 A CTIVITY VIA NFKB DEPENDENT
                     REGULATES INVASIVENESS                                        MMP-2 ACTIVITY
MODULATION               TIMP-2
MODULATION OF TIMP-2 EXPRESSION AND SECRETION
             .;
Wisniewski, P Ellert-Miklaszewska, A.; Kwiatkowska, A.; Kaminska, B.




                                                                                                                               15
Brain Tumor 2008




16
                                                                                                              Brain Tumor 2008



                              Abstracts of Poster Presentations

ADULT HUMAN MESENCHYMAL STEM CELLS AND                                EXPRESSION OF THE EGF-RECEPTOR AND NEURONAL
GLIOMA CELLS FORM A STRUCTURAL AS WELL AS A                           MARKERS IN GLIAL NEOPLASMS: A TISSUE MICROARRAY
FUNCTIONAL SYNCYTIUM IN VITRO                                         APPROACH
Albrecht V.1, Korte B.1, Buchner A.2, Riesenberg R.2,                 C. Bernreuther, T. Kamradt, L. Kolevatova, A. Kellner, J.
Mysliwietz J.3, Paron I.1, Tonn J.-C.1, Goldbrunner R.1,              Matschke, M. Glatzel
Schichor Ch.1                                                         Department of Neuropathology, University Medical Center
1Neurosurgical Clinic Grosshadern, Ludwig-Maximilians-                Hamburg-Eppendorf, D-20246 Hamburg, Germany
University, Munich, Germany, 2Laboratory for                          Pathologic diagnosis is predominantly based on conventional
Tumorimmunology, Department of Urology, Ludwig-                       haematoxylin-eosin staining but immunohistochemical analysis
Maximilians-University, Klinikum Grosshadern, Munich,                 is important in the differential diagnosis of glial neoplasms.
Germany, 3Department of Comparative Medicine, Helmholtz               Unfortunately, none of the applied markers shows absolute
Center Munich, German Research Center for Environmental               sensitivity and specificity and there is considerable variation in
Health, Neuherberg, Germany                                           the outcome between different antibodies and laboratories. In
Mesenchymal stem cells (MSCs) have come into focus as a               this study, two tissue micro arrays (TMA) displaying specimen
source for cell-based treatment strategies in glioma patients.        from ependymomas grade II and III, astrocytomas grade I-III,
The capacity of MSCs as a treatment vector is based on their          glioblastomas, and oligodendrogliomas grade II and III were
property of being recruited to malignant gliomas. This recruitment    designed to compare the applicability of different antibodies.
mechanism is driven by glioma-secreted factors which lead to          First, two different antibodies against the epidermal growth factor
accumulation of both, administered as well as bone marrow             receptor (EGFR) were applied and the results of immuno-
derived hMSCs within the tumor. Aim of our study was to evaluate      histochemical analysis were compared with the amplification of
the communication between hMSCs and gliomas in culture. Based         the EGFR gene detected by fluorescence in situ hybridization
on the finding that glioma cells as well as glioma-derived            (FISH). While both antibodies overestimated the number of gliomas
endothelial cells and hMSCs express connexins, we found an            showing an amplification of the EGFR gene as determined by
intense interaction via gap-junctional coupling. Besides this         FISH there was a significant difference in the percentage of
functional syncytium formation, we detected signs of cell fusion      positive gliomas detected by the two antibodies. These results
events, also referred to as structural syncytium. These cell-         indicate the necessity to carefully select appropriate antibodies
cellular interactions resulted in markedly enhanced migration in      if therapeutic decisions are supposed to rely on
vitro. Taken together, our data show that glioma cells as well as     immunohistochemical analysis. In a second experiment, the
endothelial cells display signs of functional as well as structural   expression of the neuronal markers neurofilament,
syncytium formation with MSCs in vitro. To our opinion, these         synaptophysin, neuronal nuclear antigen (NeuN), neural cell
basic phenomenons of cell-cellular communications within              adhesion molecule L1, and doublecortin was analysed in gliomas.
gliomas will open up entirely different views on glioma biology.      A significantly decreased percentage of astrocytomas grade I
                                                                      and ependymomas showed immunoreactivity with antibodies
                                                                      against neurofilament, synaptophysin, NeuN, and doublecortin
                                                                      when compared to astrocytomas grade II and III, glioblastomas
GENOMIC DNA METHYLATION ANALYSIS IN HUMAN BRAIN                       and oligodendrogliomas but all tested neuronal markers were
GLIOMAS                                                               detected in all examined tumour entities to a various extent.
Anna-Maria Barciszewska,1 Stanislaw Nowak,1 Iwona
Gawronska2, Miroslawa Barciszewska2
1Department of Neurosurgery and Neurotraumatology,
                                                                      TUMOR-SUPPRESSIVE ACTIONS OF HUMAN ALPHA-2-
University of Medical Sciences, Poznan, Poland, 2Institute of
                                                                      MACROGLOBULIN AT ASTROCYTIC TUMOR CELLS
Bioorganic Chemistry, Polish Academy of Sciences, Poznan,             Lindner, I.; Buchold, M.; Bigl, M.; Hutschenreuther, A.; Thal, D.
Poland                                                                Institute of Biochemistry, University of Leipzig
Despite many efforts and different approaches human brain             Malignant astrocytic gliomas such as glioblastoma are the most
gliomas remain to be a diagnostic and therapeutic challenge for       common and lethal intracranial tumors. These cancers exhibit a
modern medicine. In search for novel diagnostic methods one           relentless malignant progression characterized by widespread
can choose genetic approaches, based mostly on very popular           invasion throughout the brain, resistance to traditional and newer
PCR-based techniques. However, there is more information              targeted therapeutic approaches, destruction of normal brain
hidden in the genome namely in the cytosine methylation pattern.      tissue and certain death. Despite implementation of intensive
We decided to explore this data and use them for diagnostic           therapeutic strategies and supportive care, the median survival
purposes. We describe a simple and reliable method for detection      of that tumor has remained at 12 month over the past decade.
of 5-methylcytosine (m5C) in DNA from brain glioma tissues and        The standard therapeutic approach for treating glioblastomas is
blood samples of brain tumour patients. We present the results        a combination of radiotherapy and chemotherapy which is often
of the analysis of 250 DNA samples from brain glioma tissues,         accompanied with site effects. In the present study we investigate
half of them combined with blood samples from the same patients.      the effect of the plasma protease inhibitor, alpha2-macroglobulin
DNA was isolated, hydrolysed into nucleotides and separated           (A2M), on behaviour of astrocytoma cell in vitro. Beside its
with thin layer chromatography after labelling with 32 P.             inhibitory effect at proteases, A2M is known to be a regulatory
Chromatograms were evaluated using phosphoimager and the              protein for a number of growth factors and cytokines (1). Its
amounts of m5C calculated as a ratio of spot intensities of m5C to    receptor, LRP1, which is expressed on many tumor cells, is
m5C+C+T. The presented method of DNA analysis is very sensitive       involved in regulation of cell migration and invasion (2). To judge
and reliable. It also monitors the quality of the given sample as     the effect of A2M on tumor cells we analyzed cell proliferation,
well as isolated DNA. It seems to be not only a good diagnostic       migration, invasion, colony and spheroid formation. The final
marker for brain tumours but also differentiating low and high        results strongly indicate that lightly chemically modified A2M
grade gliomas. The R value decreases as the malignancy of             obtained from patients plasma is capable to suppress many
brain tumour increases. It also shows differences within gliomas      malignancy-associated properties of these tumor cells. Similar
grade groups. Therefore DNA methylation pattern might be a            effects were observed with antibodies directed against the
useful tool for primary diagnosis of brain tumour and a marker        extracellular domain of LRP1.Targeting LRP1 by its natural ligand
for early detection of relapse of the disease, especially because     or by engineered antibodies could provide a new treatment
the R values for the same patients are similar in their blood and     paradigm for glioblastomas.
brain tumour tissues.                                                 Acknowledgement:
                                                                      The study is supported by the Wilhelm Sander-Stiftung



                                                                                                                                     17
Brain Tumor 2008


NATURAL KILLER CELL-MEDIATED RECOGNITION OF                                 member of the Ig superfamily composed of two Ig-like domains,
HUMAN BRAIN TUMORS                                                          a transmembrane stretch and a cytoplasmic segment. CAR
Roberta Castriconi1, Alessandra Dondero1, Francesca                         isexpressed early in development on neurons and becomes
Bellora1, Antonio Daga2, Alessandro Raso3, Lorenzo                          restricted to synapse-rich layers at more advanced stages.
Moretta1,3,4, Alessandro Moretta1,4 and Cristina Bottino1,3
1Dipartimento di Medicina Sperimentale, Università degli                    Application of the fiber knob of the adenovirus, which binds to
Studi di Genova; 2 Istituto Nazionale per la Ricerca sul                    CAR, resulted in longer neurites in comparison to the untreated
Cancro; 3Istituto Giannina Gaslini; 4Centro di Eccellenza per               cell cultures. Furthermore, the formation of cell aggregates was
le Ricerche Biomediche, Università degli Studi di Genova,                   reduced by the fiber knob. By using whole-cell patch clamp
Genova, Italy.                                                              technique we analyzed the effect of the fiber knob on passive
Natural Killer (NK) cells display high cytolytic activity against           membrane properties and synaptic activity. The fiber knob was
transformed cells such as tumors and are considered suitable                able to reduce the membrane resistance (Rm). Consistently, in
candidates for adoptive immunotherapy to treat cancer patients.             CAR deficient neurons Rm was significantly higher compared to
Hematological malignances appeare to represent favorable                    wild type neurons, and application of fiber knob had no effect on
targets and preliminary studies suggest that NK cells can be                Rm on CAR-deficient neurons. Thus, CAR may influence Rm via
efficient also in eliminating solid tumors. We have analyzed the            membrane proteins with a membrane conductance. Gap junction
ability of NK cells to attack medulloblastoma and glioblastoma,             proteins form large pores with a high permeability and might
two intracranial tumors characterized by a particularly poor                therefore be candidate proteins regulated by CAR and the fiber
prognosis. We show that in spite of the absence of major                    knob. In line with this hypothesis, application of different connexin
adhesion interactions, both tumors are highly susceptible, in vitro,        blockers results in a significant increase in Rm in wild type but
to allogeneic and/or autologous activated human NK cells. The               not in CAR deficient neurons. As a consequence of the absence
NK-mediated killing involves different activating receptors and             of CAR, which seems to be essential for the regular development,
correlates with the presence of their specific ligands on tumor             synaptic connectivity was diminished, as shown by reduced
cells. Notably, NK cells could efficiently kill not only CD133-negative     frequency of inhibitory postsynaptic currents. Our results
but also CD133-positive tumor cells that, according to several              suggest that CAR might influence membrane resistance by
studies, would include Cancer Stem Cells. Finally, we analyzed              modulating the function of gap junctions. As a consequence for
the ability of both CD133-positive and CD133-negative tumor                 impaired electrical coupling in the CAR deficient neurons,
cells to secrete a large array of soluble mediators that could              formation of synaptic circuits is reduced.
promote cancer cell survival or contribute to immune evasion.


SORTING AND RELATIONSHIP BETWEEN N- AND S-TYPE                              CONTRIBUTION OF AUTOPHAGY TO DRUG-INDUCED DEATH
CELLS IN HUMAN NEUROBLASTOMA CELL LINE BY                                   OF MALIGNANT GLIOMA CELLS
                                                                            Iwona Ciechomska, Bozena Kaminska
SEDIMENTATION FIELD FLOW FRACTIONATION (SDFFF). A
                                                                            Laboratory of Transcription Regulation, Department of Cell
CELLULAR MODEL FOR CHEMOTHERAPY STUDIES.
M Cheray1,2, G. Bégaud-Grimaud1, P. Lazcoz3, J.S.                           Biology, The Nencki Institute of Experimental Biology,
Castresana3, P. J. P. Cardot1, M.O. Jauberteau2, F. Lalloué2,               Warsaw, Poland
S. Battu1.                                                                  Autophagy is a cellular process in which portions of cytoplasm
1Laboratoire de Chimie Analytique and 2Laboratoire                          and intracellular organelles are sequestered within vesicles
d’Immunologie, EA 3842, „Homéostasie Cellulaire et                          (autophagosomes) before delivery to lysosomes for degradation.
Pathologies“, Faculté de Pharmacie, Université de Limoges,                  Autophagy can be involved in cell survival or cell death. The
2 rue du Dr. Marcland 87025 Limoges cedex, France,3                         mammalian target of rapamycin (mTOR) is involved in the control
Molecular Neuro-Oncology Laboratory, Facultad de                            of cancer cell metabolism, growth and proliferation but also in
Medicina, University of Navarra, Pamplona, Spain.                           regulation of apoptosis or autophagy. Inhibition of mTOR leads to
Abstract : Neuroblastoma is the most common solid tumor of                  autophagy in malignant glioma cells. We demonstrated that
childhood. We showed that the neoplastic populations in IMR-32              cyclosporin A (CsA, a calcineurin inhibitor), UVC irradiation and
cell line are heterogeneous and variable in their state of                  adriamycin (Adr, DNA damaging drug) induced growth arrest
differentiation consisting two principal neoplastic cells: 1)               and cell death of cultured glioblastoma cells. Hallmarks of
neuroblastic (N-type undifferentiated cells); and 2) stromal (S-            apoptosis were observed after all insults, although after CsA
type differentiated cells). Heterogeneity affect treatment outcome,         they were not so displayed as after UVC and Adr treatment.
in particular the response to apoptosis induced by chemotherapy.            Moreover, CsA-induced death was accompanied the appearance
As, it seems relevant to understand the underlying process                  of numerous cytoplasmic vacuoles. Therefore, in this study we
governing changes in differentiation in order to improve treatment          investigated whether autophagy contributes to CsA-induced cell
response and patient outcome, we used SdFFF cell sorting to                 death of malignant glioma cells. Upon CsA treatment we observed
explore of the differentiation kinetic in this cell line. SdFFF permitted   several features characteristic of autophagy: development of
the cell sorting of 2 different N-phenotypes and the understanding          the acidic vesicular organelles, association of microtubule-
of the cell population dynamism. The first N-phenotype forms a              associated protein light-chain 3 (LC3) with autophagosome
pool of quiescent cells. the second one is able to proliferate              membrane, and a marked increase in the level of LC3-II expression
(BrdU incorporation) and also to differentiate into adherent S-             in malignant glioma cells. We found that CsA treatment affects
type cells (N-CAM+). Thanks to SdFFF, we isolated the second                mTOR/p70S6 kinase pathway decreasing p70S6K
N-type phenotype which have characteristics of malignant cells              phosphorylation. Further, the inhibitory effect of CsA on the
(high expression of nestin) and transiently expressed the PSA-              phosphorylation of 4EB-P1 (eukaryotic initiation factor 4E-binding
N-CAM which could play a role in the metastatic properties of               protein 1) and S6 ribosomal protein (two downstream target
these cells. Then, as these phenotypes could be routinely                   molecules of p70S6K) was observed. The presented results
prepared by SdFFF, they could be used to study their selective              demonstrate that CsA induces autophagy-associated cell death
sensitivity to apoptosis and differentiation therapies.                     in malignant glioma cells via inhibition of mTOR/p70S6K pathway.
                                                                            It shows a possibility to induce nonapoptotic autophagic cell
                                                                            death in malignant gliomas bearing numerous defects in apoptotic
MODULATION OF NEURONAL MEMBRANE PROPERTIES BY                               pathway.
THE COXSACKIEVIRUS-ADENOVIRUS RECEPTOR
Cholewa J., Jüttner R. and F. G.Rathjen
Max Delbrück Center for Molecular Medicine, Berlin
The coxackie virus and adenovirus receptor (CAR) was originally
identified as a cell surface protein enabling both viruses to interact
with cells. Besides this pathological role as a virus receptor, the
physiological function in the CNS is largely unknown. CAR is a



18
                                                                                                                Brain Tumor 2008


SPECIFIC MORPHOLOGY AND AUTOFLUORESCENCE                               new molecular targeted therapies. We used the microarray
IDENTIFY SELF-RENEWING AND TUMOR-INITIATING                            technique to compare the transcriptional profiles of 5 H/C PA and
GLIOMA CELLS                                                           6 cerebellar PA. Validation of the microarray results and
Virginie Clément°, Denis Marino°, Cristina Cudalbu+, Vladimir          comparison of PA with normal developing tissue was done by
Mlynarik+, Marie-France Hamou§, Nicolas de Tribolet°, Pierre-
Yves Dietrich*, Rolf Gruetter+, Monika Hegi§ and Ivan                  quantitative RT-PCR and immunohistochemistry. Results
Radovanovic°                                                           demonstrate that cerebellar and H/C PA are two genetically distinct
Centre Romand de Neurochirurgie and *Dpt. of Oncology,                 and topography-dependent entities. Numerous genes
Geneva University Hospitals and University of Geneva,                  upregulated in H/C PA also increased in the developing chiasm,
Geneva, Swizterland. +Laboratory for Functional and                    suggesting that developmental genes mirror the cell of origin
Metabolic Imaging, Ecole Polytechnique Federale de                     whereas migrative, adhesive and proliferative genes reflect
Lausanne, Lausanne, Switzerland. § Laboratory of Tumor                 infiltrative properties of H/C PA. Of particular interest, NOTCH2, a
Biology and Genetics, Centre Romand de Neurochirurgie,                 gene expressed in radial glia and involved in gliomagenesis,
Centre Hospitalier Universitaire Vaudois and University of             was upregulated in H/C PA. In order to find progenitor cells that
Lausanne, Lausanne, Swizterland                                        could give rise to H/C PA we performed a morphological study of
Human gliomas are primary neoplasms of the CNS that show               the H/C region and identified, in the floor of the third ventricle, a
different grades of local aggressiveness and display various           unique population of vimentin- and GFAP-positive cells highly
cell phenotypes of glial lineages but also of less differentiated      suggestive of radial glia cells. Therefore, PA of the H/C region
neural stem/progenitor cells (Wechsler-Reya et al, 2001; Read          should be considered as a distinct entity which probably
et al, 2006; Sanai et al, 2005). Although the exact cellular origin    originates from a unique population of cells with radial glia
of gliomas remains unclear, it is proposed that a small fraction of    phenotype.
cancer cells constitutes a unique reservoir of glioma initiating
cells (GICs) controlling tumor growth (Galli et al, 2004; Hemmati
et al, 2003). GICs were prospectively identified as CD133-             IDENTIFICATION OF CANCER STEM CELLS IN A MOUSE
expressing cells (Singh et al, 2003, 2004). However, CD133             MODEL OF MEDULLOBLASTOMA AND MOLECULAR
might not be sufficient to identify all GICs as CD133- cells derived   COMPARISON WITH NORMAL NEURAL STEM CELLS.
                                                                       Daniela Corno1, Barbara Cipelletti1, Mauro Pala2, Letterio
from some glioma types have comparable self renewal and                Politi3, Alessandro Bulfone2 and Rossella Galli1.
tumorigenic properties (Wang et al, 2007; Zheng et al, 2007;           1Stem Cell Research Institute (SCRI) — San Raffaele
Beier et al, 2007; Odgen et al, 2008) For further comprehension        Scientific Institute, Via Olgettina 58, Milan, Italy, 20132,
of brain tumor cellular hierarchies it is therefore necessary to       2bio))flag Srl, Scientific Park of Sardinia, Pula, Cagliari, Italy,
develop alternative or complementary selection strategies to           3Neuroradiology Unit – San Raffaele Scientific Institute, Via
identify, isolate and characterize GICs. Here, we show that the        Olgettina 60, Milan, Italy, 2013220129;
combination of a distinctive morphology and autofluorescence           Medulloblastomas (MBs) are the most common malignant brain
signals upon 488nm laser excitation allows the identification of a     tumors in childhood and are thought to arise from cerebellar
subpopulation of cultured or prospectively isolated glioma cells       progenitors that undergo malignant tranformation. In particular,
with tumor- initiating and long term self-renewing capacities.         the desmoplastic MB variant seems to derive from external
Furthermore, glioma cell autofluorescence correlates with              granule layer (EGL precursors, whereas the classic variant
enhanced expression of stemness genes, higher metabolic                appears to originate from IV ventricle progenitors. Cancer stem
activitiy and specific cell cycle profile. Complementing CD133 to      cells (CSCs), capable to sustain tumor growth, progression and
our phenotypical methodology yields a subpopulation with               recurrence, have been identified in different human brain tumors,
superior self-renewal ability than with one or the other method        including MBs. In this study, we focused on the biological and
alone. Taken together, our findings suggest that self-renewing         molecular characterization of CSCs, obtained from MBs
and tumor-initiating glioma cells have a distinct morphology and       developing spontaneously in patched mutant mice. We isolated
autofluorescent phenotype that reflects a higher metabolic state.      several lines of MB CSCs, by culturing tumor specimens under
Our unbiased phenotypic selection thus provides a unique               the standard conditions used for normal neural stem cells (NSCs).
opportunity to investigate the tumor initiating potential of a given   These MB cell lines displayed all the features that qualify them as
subpopulation or a single tumor cell.                                  CSCs, i.e. extensive proliferation, long-term self-renewal,
                                                                       aberrant differentiation, and, most importantly, tumor initiating
                                                                       ability. To assess if a lineage relationship could be revealed
PILOCYTIC ASTROCYTOMA OF THE OPTIC PATHWAY: A                          between MB CSCs and normal NSCs by the identification of a
TUMOR DERIVING FROM RADIAL GLIA CELLS WITH
                                                                       common molecular signature, we isolated NSCs from the EGL,
SPECIFIC GENE SIGNATURE
Aurélie Tchoghandjian1*, Carla Fernandez1,2*, Carole Colin1,           the IV ventricle and the subventricular zone (SVZ) of post-natal
Ikbale El Ayachi1, Brigitte Voutsinos-Porche1, Frédéric Fina3,         day 7 wild-type mouse brain, and subjected them to gene
Didier Scavarda4 , Marie-Dominique Piercecchi-Marti 2,                 expression profiling together with MB CSCs. Microarray-based
Dominique Intagliata1, L’Houcine Ouafik1, Caroline Fraslon-            analysis showed that each MB CSC line displayed a typical
Vanhulle 5 and Dominique Figarella-Branger1, 2                         transcriptional profile. When compared with normal NSCs, MB
1UMR911-CRO2, Faculté de Medecine Timone, Université de                CSCs appeared to share more molecular determinants with
la Méditerranée, Marseille, F-13000, France; 2Assistance               cerebellar NSCs than with SVZ NSCs. Thus, by exploiting the
Publique-Hôpitaux de Marseille, CHU Timone, Service                    link existing between the biology of the stem cell component in
d’Anatomie Pathologique et de Neuropathologie, Marseille, F-           MBs and the physiological development of the cerebellum, it might
13000, France; 3Laboratoire de Transfert d’Oncologie                   be possible to identify still unknown pathogenetic mechanisms
Biologique, APHM, Faculté de Médecine Nord, Marseille, F-              and new candidate targets useful to develop novel and selective
13000, France; 4Assistance Publique-Hôpitaux de Marseille,             treatments for MBs.
CHU Timone, Service de Neurochirurgie, Marseille, F-13000,
France; 5Sanofi-Aventis Recherche et Développement,
Oncology Department, Sanofi-Aventis, Vitry-sur-Seine, F-               EFFECTS OF SUNITINIB ON TUMOR HEMODYNAMICS AND
94400, France.                                                         DELIVERY OF CHEMOTHERAPY
* These 2 authors have equally contributed to the work.                M. Czabanka1 , M. Vinci2, F. Heppner4, A. Ullrich3, P.
Pilocytic astrocytomas (PA) are WHO grade I gliomas that occur         Vajkoczy1
predominantly in childhood. They share features of both astroglial     1Department of Neurosurgery, Universitätsmedizin Charitè,
and oligodendroglial lineages. These tumors affect preferentially      Berlin, 2Department of Neurosurgery, Faculty for Medicine
the cerebellum (benign clinical course) and the optic pathway,         Mannheim, University of Heidelberg, 3Max Planck Institute
especially the hypothalamo-chiasmatic (H/C) region (poor               for Biochemistry, Martinsried, 4Department of
prognosis). Understanding the molecular basis responsible for          Neuropathology, Universitätsmedizin Charitè, Berlin
the aggressive behaviour of H/C PA is a prerequisite to set up         Current clinical protocols favour a combination of anti-angiogenic/




                                                                                                                                        19
Brain Tumor 2008


anti-vascular compounds with classical chemotherapy. However,           differentiation. Originally identified on the basis of their mutational
it remains unclear to what extent an anti-angiogenic/anti-vascular      phenotypes in stem-cell maturation, mir-125 and let-7 are strongly
therapy influences the delivery of chemotherapy. Therefore, the         induced during neural differentiation of embryonic stem (ES)
aim of the present study was to characterize the effects of the         cells and embryocarcinoma (EC) cells. We report that embryonic
anti-angiogenic tyrosine kinase inhibitor Sunitinib on tumor            neural stem (NS) cells express let-7 and mir-125, and investigate
microhemodynamics and delivery of chemotherapy. SF126 tumor             post-transcriptional mechanisms contributing to the induction of
cells were implanted subcutaneously into nude mice and were             let-7. We demonstrate that the pluripotency factor Lin-28 binds
analysed repeatedly by intravital microscopy. Treatment with            the pre-let-7 RNA and inhibits processing by the Dicer ribonuclease
Sunitinib was initiated 7 days after implantation. In order to assess   in ES and EC cells. In NS cells, Lin-28 is downregulated by mir-
the effects of Sunitinib on tumor vasculature and hemodynamics          125 and let-7, allowing processing of pre-let-7 to proceed.
we analysed total and functional vessel densities, microvascular        Suppression of let-7 or mir-125 activity in NS cells led to
diameter and microvascular blood flow rate. In order to study the       upregulation of Lin-28 and loss of pre-let-7 processing activity,
delivery of chemotherapy, autofluorescent doxorubicin was               suggesting that let-7, mir-125 and lin-28 participate in an
systemically administered and its vascular delivery to the tumor        autoregulatory circuit that controls miRNA processing during NS-
tissue was quantified. Histological analysis included endothelial       cell commitment.
cell proliferation, pericyte coverage of tumor vessels and tumor
cell proliferation. Sunitinib significantly suppressed tumor growth
by both anti-vascular and anti-angiogenic effects. However, a           BLOCKING OF MICROGLIAL INFILTRATION AND
number of tumor vessels escaped anti-angiogenic therapy.                ACTIVATION IN TRANSPLANTED GLIOMAS WITH
Interestingly, in these surviving blood vessels Sunitinib treatment     SYSTEMICALLY APPLIED CYCLOSPORIN A
resulted in an increased microvascular blood flow rate resulting        SIGNIFICANTLY REDUCES TUMOR GROWTH
in an improved delivery of chemotherapy via these blood vessels.        Gabrusiewicz, K.; Porycka, M.; Frankowska, M.; Kaminska, B.
Besides its potent anti-angiogenic and anti-vascular efficacy           Laboratory of Transcription Regulation, Department of Cell
Sunitinib treatment results in improved microhemodynamics and           Biology, Nencki Institute, Warsaw, Poland
blood flow in tumor blood vessels that escape therapy leading to        Recent evidence suggest that brain tumour-infiltrating microglia/
an improved vascular delivery of chemotherapy. These results            macrophages support glioma growth and invasion. Understanding
provide the basis for a potential chemosensitizing effect of            of molecular mechanisms responsible for pathogenesis of glioma
Sunitinib.                                                              and their interactions with microenvironment permits to identify
                                                                        microglia-glioma interactions as a novel therapeutic target. We
                                                                        have demonstrated that cyclosporin A (CsA) affects growth/
BOTH CD133 POSITIVE AND CD133 NEGATIVE                                  survival of cultured glioblastoma cells, interferes with glioma-
MEDULLOBLASTOMA CELL LINES EXPRESS LIGANDS FOR                          microglia interactions and impairs tumorigenicity of intracranial
TRIGGERING NK RECEPTORS AND ARE SUSCEPTIBLE TO                          gliomas (Sliwa et al. 2007). In the present study we investigated
NK-MEDIATED CYTOTOXICITY                                                efficacy and mechanisms mediating antitumor effects of CsA in
Dondero1, A.; Castriconi1, R.; Negri2, F.; Bellora1, F.; Nozza2,        vivo, with particular attention to drug influence on density and
P.; Carnemolla3, B.; Raso2, A.; Moretta1,2,4, L.; Moretta1,4 , A.;      morphology of brain macrophages and level of pro/anti-
Bottino1,2, C.                                                          inflammatory cytokines. EGFP-GL261 glioma cells were injected
1Dipartimento di Medicina Sperimentale, Università degli                into the striatum of C57BL/6 mice and tumor-bearing mice received
Studi di Genova, Via L.B. Alberti 2, 16132 Italy; 2Istituto             i.p. CsA (2 or 10 mg/kg) every 2 days starting from the 2nd or the
Giannina Gaslini, L.go G. Gaslini 5, 16147 Genova, Italy;               8th day after implantation. We demonstrate that CsA-treated
3Istituto Nazionale per la Ricerca sul Cancro, L.go Rosanna             mice had significantly smaller tumors than control mice. When
Benzi 10, 16132 Genova, Italy; 4Centro di Eccellenza per le             the treatment was postponed to 8th day, only the higher dose of
Ricerche Biomediche, Università degli Studi di Genova, V.le             CsA was effective causing 66 % tumor volume reduction. CsA-
Benedetto XV, 16132 Genova, Italy.                                      treated mice showed a diminished number of tumor-infiltrating,
Medulloblastoma represent the most common malignant                     amoeboid brain macrophages. TUNEL staining revealed a DNA
intracranial tumor of childood characterized by a dramatic              fragmentation within infiltrating macrophages and glioma cells
prognosis. Thus adoptive cellular immunotherapy could be utilized       after CsA treatment. Immunosuppressant at given concentration
in the treatment of medulloblastoma together with neurosurgical,        (<10 µM) did not affect proliferation or viability of cultured EGFP-
radiotherapy and chemothearapy. In this study we have analyzed          GL261 glioma cells. Pro/anti-inflammatory cytokine levels were
the ability of the activated human Natural Killer cells to kill this    determined in extracts from tumor-bearing hemisphere using
tumor. We showed that medulloblastoma is susceptible in vitro to        FlowCytomix immunoassay. Only IL-10 and GM-CSF levels were
NK- mediated killing and clarified the receptor-ligand interactions     increased in tumor-bearing hemisphere in comparison to naive
involved in this killing. In particular we demonstrated that the NK-    controls and CsA treatment reduced significantly IL-10 and GM-
mediated killing of medulloblastoma is due to the cooperation of        CSF levels. Thus targeting of cytokine production, macrophage
different activating NK receptors including NKp46, NKp30, DNAM-         infiltration and interactions with glioma may be a novel antitumor
1 and NKG2D. Moreover medulloblastoma expressed a number                strategy.
of tumor-associated molecules including CD146 and CD133,
considered a marker for cancer stem cells. Remarkably both
CD133-positive and CD133-negative cell lines were susceptible           CARNOSINE INHIBITS GROWTH OF CELLS DERIVED FROM
to lysis. Tumor cells also expressed molecules that are currently       HUMAN GLIOBLASTOMA MULTIFORME AND IN AN
used as diagnostic tools for neuroblastoma cell identification. In      ANIMAL MODEL OF HER2/NEU BY INTERFERENCE WITH
particular, B7-H3 was expressed by all the medulloblastoma cell         ANAEROBIC GLYCOLYSIS
lines analyzed, while the presence of GD 2 and NB84 was                 Gaunitz, F.2; Renner, C.1; Seyffarth, A.2; Asperger, A.1;
restricted to given cell lines and/or marked a defined tumor cell       Zemitzsch, N.2; Gebhardt, R.2; Meixensberger, J1.
subset.                                                                 1 Klinik und Poliklinik für Neurochirurgie, 2Institut für
                                                                        Biochemie, Medizinische Fakultät der Universität Leipzig,
                                                                        Leipzig, Germany
A FEEDBACK LOOP COMPRISING LIN-28 AND LET-7                             Glioblastoma multiforme is the most aggressive and the most
CONTROLS PRE-LET-7 MATURATION DURING NEURAL                             frequent primary brain tumor. Despite a number of therapeutic
STEM-CELL COMMITMENT.                                                   advances in recent years, prognosis remains poor. The median
Rybak A, Fuchs H, Smirnova L, Brandt C, Pohl EE, Nitsch R,              duration of survival after first biopsy is close to 40 weeks and
Wulczyn FG.                                                             there is currently no pharmacological treatment available that
Institute of Cell and Neurobiology, Center for Anatomy,                 significantly increases this time. We recently presented evidence
Charité - Universitätsmedizin Berlin, Berlin, Germany.                  that the naturally occurring dipeptide carnosine inhibits growth
miRNA populations, including mammalian homologues of lin-4 (mir-        of cells from the cell line T98G that was derived from human
125) and let-7, undergo a marked transition during stem-cell



20
                                                                                                                 Brain Tumor 2008


glioblastoma multiforme as well as that of primary cells derived       homogenates increased with malignancy. In vitro, CXCR7 was
from human glioblastoma [1]. Biochemical and cytochemical              highly expressed in all glioma cell lines investigated whereas
investigations now revealed that treatment with carnosine              CXCR4 was only scarcely transcribed on 1 of 8 glioma cell lines.
reduces ATP production in these cells by interference with             On glioma cells, CXCR7 was upregulated by transforming growth
anaerobic glycolysis. In addition, we demonstrate that carnosine       factor-β (TGF-β), but not by its ligand, growth factors or pro-
is able to retard tumor growth in vivo in a Her2/Neu mouse model.      inflammatory cytokines. Stimulation of glioma cells (CXCR7-
Since carnosine is known to be able to penetrate the blood-brain       positive, CXCR4- and CXCR3-negative) by CXCL12 or CXCL11
barrier and appears to be non-toxic to normal tissue this              induced transient phosphorylation of extracellular-signal regulated
substance may be a candidate for a therapeutic agent that may          kinases Erk1/2 indicating that the receptor is functionally active.
reduce proliferation of neoplastic cells and especially in cases       Whereas proliferation was little influenced, chemokine stimulation
of glioblastoma multiforme.                                            prevented Camptothecin-induced apoptosis. Thus, CXCR7 and
References                                                             not CXCR4 is the major receptor for CXCL12 (and CXCL11) in
1. Renner C, Seyffarth A, de Arriba SG, Meixensberger J,               astrocytomas / gliomas and mediates resistance to apotosis.
Gebhardt R, Gaunitz F. Carnosine inhibits growth of cells
isolated from human glioblastoma multiforme. International
Journal of Peptide Research and Therapeutics 2008; 14(2):              GATA-2 AND -3 ARE HIGHLY EXPRESSED IN
127-135                                                                NEUROBLASTOMA OF FAVORABLE SUBTYPE
                                                                       Victoria Hoene1, Matthias Fischer2, Anna Ivanova1, Frank
                                                                       Berthold2, and Christof Dame1
                                                                       1Department of Neonatology, Campus Virchow-Klinikum,
BMP7 RELEASE FROM ENDOGENOUS NEURAL
PRECURSORS ATTENUATES TUMORIGENICITY OF GLIOMA                         Charité – Universitätsmedizin Berlin and 2Department of
INITIATING CELLS                                                       Pediatric Oncology and Hematology, University Children’s
Chirasani SR1, Sternjak A2, Wend P3, Momma S4, Herold-                 Hospital of Cologne
Mende C5, Besser D3, Synowitz M6 & Kettenmann H1 Glass                 GATA transcription factors comprise a family of six zinc finger
R1,                                                                    proteins that regulate cell differentiation and proliferation in normal
Cellular Neuroscience, 2 Cellular Immunology and                       development. However, some family members have also been
3Embryonic Stem Cells, Max Delbrück Centre for Molecular               associated with tumors. We recently found that GATA-4 is
Medicine (MDC), Berlin,Germany. 4 Edinger Institute                    expressed in human neuroblastoma with a higher expression
Frankfurt/M., Germany. 5Neurosurgery, University                       level in more aggressive tumor subtypes while absent in the
Heidelberg, Germany. 6Neurosurgery, Charité, Berlin,                   developing sympathetic nervous system (SNS). We sought to
Germany                                                                analyze the role of GATA-2 and GATA-3 in this setting since in
Abstract: We have established previously that endogenous               contrast to GATA-4, they are expressed during normal
neural precursor cells (NPCs) are attracted to experimental brain      development of the SNS and essential for the generation and
tumors (gliomas) in large numbers and reduce tumor size. We            differentiation of sympathetic neurons. Herein, we found nuclear
now report that NPCs attenuate the tumorigenic potential of glioma     GATA-2 and GATA-3 localization in the human neuroblastoma
initiating cells via BMP7 release. Using an in vivo model we show      cell line SH-SY5Y as well as in human neuroblastoma specimens.
that glioma-associated PSA-NCAM-positive NPCs express BMP7,            Microarray analyses showed significantly higher GATA-2
but not BMP2 or BMP4. BMP7 is the pre-dominant BMP in                  expression levels in MYCN-nonamplified (n=218) vs. MYCN-
neuroshere cultures and is exclusively expressed by PSA-               amplified (n=32) tumors (p<0.001). GATA-3 revealed a similar
NCAM-positive NPCs, but not by differentiating or tumor cells.         trend but not a significant association. Correspondingly, both
Moreover, NPCs constitutively release BMP7. Neurosphere                factors were elevated in low risk (n=169) compared to high risk
conditioned media or recombinant BMP7 were applied to murine           (n=82) tumors (p<0.001 for GATA-2; p=0.001 for GATA-3)
or human glioma and, BMP-receptor dependently, reduced the             according to a highly accurate clinico-genetic classification based
CD133-positive population, attenuated expansion, sphere                on the tumors’ gene expression patterns. Interestingly, friend of
formation and cell viability and induced markers of differentiation.   GATA (FOG)-2 showed the same expression pattern as GATA-
Likewise, NPCs caused smad1/5/8 signalling, ID1 and ID2                2 and -3. In summary, GATA 2 and -3 as well as their cofactor
expression and Olig2 down-modulation in human CD133+ glioma.           FOG-2 are highly expressed in favorable neuroblastoma
Importantly, exposure of CD133+ murine glioma cells to                 subtypes, in contrast to GATA 4. Ongoing studies will elucidate
recombinant BMP7 prior to intracerebral implantation of 100 viable     the function of GATA transcription factors in neuroblastoma.
cells in a mouse model significantly prolonged survival as
compared to injection of untreated control cells. Overall, our
results show that NPC-released BMP7 attenuates tumor initiation        COMPREHESIVE (MOLECULAR) CYTOGENETIC
by CD133+ glioma cells and point out a role for BMPs as paracrine      CHARACTERIZATION OF RARE INTRACRANIAL TUMORS
tumor suppressors in the CNS.                                          Holland, H.1; Krupp, W.2; Koschny, R.3; Livrea, M.4; Schober,
                                                                       R.5; Meixensberger, J.2; Ahnert, P.6
                                                                       1 Translational Centre for Regenerative Medicine, University
EXPRESSION AND FUNCTION OF THE CHEMOKINE                               of Leipzig, Germany; 2 Clinic of Neurosurgery, University of
RECEPTOR CXCR7 IN HUMAN GLIOMAS                                        Leipzig, Germany; 3 Dept. of Internal Medicine, University of
Hattermann, K.*; Held-Feindt, J.#; Lucius, R*; Mentlein, R.*           Heidelberg, Germany; 4 Fraunhofer Institute for Cell Therapy
*University of Kiel, Department of Anatomy, 24098 Kiel,                and Immunology, Leipzig, Germany; 5 Dept. for
Germany, # University of Schleswig-Holstein Medical Center,            Neuropathology, University of Leipzig, Germany; 6 Institute for
Department of Neurosurgery, 24105 Kiel, Germany                        Medical Informatics, Statistics and Epidemiology, University
The chemokine CXCL12 / SDF-1 and its receptor CXCR4 play a             of Leipzig, Germany
major in tumor invasion, proliferation and metastasis. Recently,       Chromosomal aberrations play an important role in tumor
CXCR7 was identified as a novel receptor for CXCL12 and                formation, as predictors of clinical outcome and response to
CXCL11 / I-TAC. Both chemokines were detected abundantly in            therapy. Only a few publications report about (molecular)
human astrocytomas and gliomas; however, CXCR4 has only a              cytogenetic cytogenetic investigations of rare brain tumors.
restricted distribution on distinct glioma cells. Therefore, we        Therefore, we performed the first comprehensive cytogenetic
investigated the expression of CXCR7 in solid human                    analyses in esthesioneuroblastoma, adult medulloblastoma,
astrocytomas and glioma cell line by quantitative RT-PCR, Western      atypical meningioma, and hemangiopericytoma using trypsin-
blot and immunohistochemistry. We found that CXCR7 is highly           Giemsa staining (GTG-banding), multicolor fluorescence in situ
expressed on tumor endothelial cells, tumor-invading microglial        hybridization (M-FISH) and molecular karyotyping using single
cells / monocytes and astrocytoma / glioma cells in situ. CXCL12       nucleotide polymorphism array (SNP-A). Structural chromosomal
is often co-localized with CXCR7 indicating that most of the ligand    aberrations were found, predominantly located on chromosomes
is bound to this receptor in situ. CXCR7-transcription in              2q,6q,21q,22q (esthesioneuroblastoma), 4q,9q,10q,11p,20q



                                                                                                                                          21
Brain Tumor 2008


(adult medulloblastoma), 6q,8q,10q,12p (atypical meningioma),           cells in a non-destructive manner opens the possibility to
and 8q,10q (hemangiopericytoma). Novel, so far not described            characterize cells on a molecular level without labels or an
chromosomal aberrations were detected: deletions                        irreversible treatment. The objective of the present study was to
del(2)(q37),del(21)(q22) [esthesioneuroblastoma], translocations        establish the conditions for infrared spectroscopic imaging of
t(4;11)(q25;p15),t(9;20)(p23;p12) [adult medulloblastoma],              neural stem cells in a cell culture. This is experimentally
t(8;19)(q24;q13), t(10;16)(q22;q12.1) [atypical meningioma], and        challenging, but offers advantages such as the reduction of
(partial) trisomy 8 [hemangiopericytoma]. For the first time, SNP-      artefacts due to fixation. Furthermore, the application may
A karyotyping revealed partial uniparental disomy on chromosomal        facilitate in-situ monitoring of spectral changes with respect to
regions 1q, 9q (adult medulloblastoma), 1p31.1, 2p16.1,                 different phenotypes. Cell cultures of murine neural stem cells
2q23.3,6q14.1, 6q21, 9p21.1, 10q21.1, and 14q23.3 (atypical             were characterized using infrared spectroscopic imaging in
meningioma). Our study underlines the necessity to apply                conjunction with multivariate chemometric data analysis. Principal
complementary methods for a comprehensive cytogenetic                   component analysis and linear discriminant analysis were
analysis of tumor genomes.                                              performed on the spectroscopic data sets to distinguish different
                                                                        cell phenotypes without labeling. Significant spectral changes
                                                                        were observed in the composition of proteins and DNA content.
DEFINING IN VITRO CONDITIONS FOR ENRICHMENT OF                          Different types of neural cells such as astrocytes, neurons and
STEM-LIKE CELL POPULATION IN PRIMARY HUMAN BRAIN                        stem cells were classified according to the gold standard of
TUMOR CULTURES                                                          immunohistochemical labeling. The results show that infrared
U. Kahlert, J. Maciaczyk, G. Nikkhah                                    spectroscopic imaging contributes new insights that are not
Dept. of Stereotactic Neurosurgery, Laboratory Molecular                available by common histochemical methods.
Neurosurgery, University Hospital Freiburg, Neurocentre,
Freiburg, Germany
Inhibition of mutated mechanisms (like NOTCH or SHH) in                 DEVELOPMENT OF A 3D IN VITRO MODEL FOR
glioblastoma derived cell lines or changes in the activity of           LONGITUDINAL OBSERVATION OF CELL AGGREGATES
aberrant signal transduction (i.e. via Bone Morphogenetic               COMBINING HIGH FIELD MAGNETIC RESONANCE IMAGING
Proteins) resulted in significant reduction of brain tumor stem cell    (MRI) WITH OPTICAL TECHNIQUES
population as well as reduced in vitro proliferation and tumor          K. Kruttwig1, C. Brueggemann1, E. Kaijzel2, S. Vorhagen1, T.
formation potential in vivo. This interventions in various levels of    Hilger3, C. Löwik2, M. Hoehn1
biochemical pathways could lead to the possibly of developing           1In-vivo-NMR-Laboratory, MPI for Neurological Research,
novel approaches in brain tumor therapy improving chemo- and            Cologne, Germany, 2Leids Universitair Medisch Centrum,
radio sensitivity. Our project is focused on defining the in vitro      Leiden, The Netherlands, 3University Clinics Cologne, Dept.
cell culture conditions leading to enrichment in stem-like cell         of Radiation Therapy, Cologne, Germany
population. Freshly resected high grade glial tumors were               Molecular imaging is a very sensitive and versatile tool for the
carefully dissociated to single cell suspension and subsequently        detection of specific groups of cells and, also, for the cellular
propagated as spherical cell aggregates under the influence of          characteristics in tumour biology. For an intensive and longitudinal
mitogens. Brain tumor cultures from different donors were               characterisation of glioma cell proliferation and for large scale in
characterised by growth kinetics, gene profiling and                    vitro screening purposes we have developed a 3D agarose-
differentiation potential. Two media-compositions (DMEM/F12 and         based in vitro model with the focus on the detectability and
Neurocult® based) and oxygen levels (21% and 3%) were                   coregistration in different imaging modalities. The combination of
analysed. Applying a longitudinal analysis with MTS assay, a            optical imaging, phase-contrast microscopy and high-field MRI
suppressed cell growth in hypoxic conditions was demonstrated           provides the possibility to study, in detail with complementary
as compared to significantly faster proliferation in DMEM/F12           imaging techniques, the behaviour of small cellular tumour-
based media. FACS analysis for CD 133- a marker of stem-like            aggregates. Here we present the investigation of the human
cells- showed significant increase of this population following         glioblastoma cell line Gli36dEGFR, genetically modified to express
cultivation in low oxygen combined with Neurocult® based media.         the fluorescent protein mCherry. Furthermore, the cells were
DMEM/F12 culture media seem to boost the proliferation of fast          labelled with a MRI contrast agent, producing a pronounced T2*-
dividing progenitor cell population as revealed by a lower CD           weighted contrast. Through the combination of 2D optical imaging
133+ level. Variations in differentiation potential (10 days in DMEM/   technology and 3D MRI, the position of individual aggregates and
F12 based media without mitogens) and gene expression profiles          their corresponding fluorescent light intensity can be detected
for oncogenic genes, SHH, WNT and stemness were assessed                separately in vitro studies. With this model system aggregates of
histochemically and by semi-quantitative PCR.                           a wide range of sizes can be investigated in parallel. Together
                                                                        with the multimodal imaging approach it is possible to localize cell
                                                                        aggregates in space, quantify the cell number and follow their
INFRARED SPECTROSCOPIC IMAGING: TOWARDS A                               corresponding proliferational activity over time. To assess further
METHOD FOR LABEL FREE CHARACTERIZATION OF                               application potential of this model system for live cell monitoring
NEURAL STEM CELLS                                                       we investigated the response of cell aggregates to gamma-ray
Matthias Kirsch (1), Saskia Küchler (1), Andreas Hermann (2),           exposure, thus demonstrating the usefulness of the system for
Edmund Koch (3), Reiner Salzer (4), Gabriele Schackert (1),             in cancer treatment evaluation strategies.
Gerald Steiner (3)
1 - Klinik und Poliklinik für Neurochirurgie,
Universitätsklinikum Carl Gustav Carus, 2 - Klinik und
                                                                        GLOBAL GENE EXPRESSION PROFILING REVEALS
Poliklinik für Neurologie, Universitätsklinikum Carl Gustav             DISTINCT GENOMIC RESPONSES IN PRIMARY
Carus, 3 - Clinical Sensoring and Monitoring,                           MICROGLIAL CULTURES
Universitätsklinikum Carl Gustav Carus, 4 - Institut für                Maciej Lipko, Michal Dabrowski, Bozena Kaminska
Analytische Chemie, Technische Universität Dresden                      Department of Cell Biology, The Nencki Institute of
Rapid and reliable discrimination of cell types and their temporal      Experimental Biology, Warsaw, Poland
changes is a crucial task in cell biology. In particular, neural stem   Microglia are multifunctional immune cells of the brain executing
cells are currently in the focus of research since they have the        various functions in response to pathological insults. Malignant,
potential to treat neurodegenerative disease such as Alzheimer          infiltrative gliomas recruit microglia and transform into tumour-
disease, chronic stroke and multiple sclerosis. These possible          supportive cells with no features of inflammation. We
applications require methods for monitoring the development and         hypothesized that microglia may be differently activated under
differentiation of the cells. However, available techniques, based      pathological conditions. In contrast to lipopolysaccharide (LPS)
on an irreversible treatment of the cells, do not allow a sensitive     induced activation, involving all MAP kinases and production of
label free characterization under in situ conditions. The potential     inflammatory mediators, glioma derived factors induce p38 MAPK
of optical spectroscopic methods such as Raman and infrared             and JNK signalling, release of anti-inflammatory cytokines,
spectroscopy to assess the overall molecular composition of



22
                                                                                                                  Brain Tumor 2008


stimulate motility and invasiveness of glioma cells. Using               of fluorescently labeled F8-SIP (n=5 per group). Host vasculature
Affymetrix arrays, we characterized gene expression in primary           of mice without tumor bearing DSCs were used as control group.
microglial cultures exposed to LPS or glioma derived factors.            Results: F8-SIP binds specifically to tumor vessels reaching its
Bioinformatics and Gene Ontology analysis revealed differences           maximum binding capacity 4 hours after injection (t0 h:26,12±10,89
in extent and profiles of expression of genes encoding cytokines/        vs. t4 h:91,46±6,19; p<0,05). In control vasculature no binding
chemokines and transcription regulators suggesting distinctive           was observed. Extravasation of F8-SIP to tumor interstitium
genomic responses and pathway specific signalling. The most              was observed reaching its maximum 4 hours after injection (t0
discriminative feature was inhibition of interferon signalling           h: 25,92±3,47 vs. t4 h: 60,41±7,77; p<0,05). Microvascular binding
pathway in glioma exposed microglia. The expression of other             was flow-dependent with significantly increased binding in high
mediators of inflammation (COX-2, iNOS, Il-1b), highly upregulated       flow blood vessels (>60 nl/sec) compared to low flow blood
after LPS treatment left unaffected in microglia treated with glioma     vessels (<20 nl/sec) at t2 h by 44% and at t4 h by 22 %,
derived factors. Unlikely, genes involved in degradation/                respectively. F8-SIP binding occured preferentially in angiogenic
remodelling of extracellular matrix were regulated in similar way        sprouts (AS) compared to remaining tumor vasculature (RTV) 2
by LPS and glioma derived factors. Differences in gene                   hours after injection (AS: 115,14±12,38 vs. RTV: 77,86±11,25;
expression in differently stimulated microglia were confirmed by         p<0,05). Conclusion: F8-SIP represents a useful tool to specifically
qPCR on independently derived cultures. We demonstrate that              target tumor microvessels. Microvascular binding occurs in a
while LPS or glioma released factors induce similar                      time- and blood flow dependent manner with preferential binding
transformation into amoeboid, motile microglia, “inflammatory”           sites. Our results provide biodistribution characteristics that might
microglia differs functionally from glioma-associated microglia.         be used for future clinical applications.
Identification of distinctive features and signalling pathways will
allow to target specific functions of microglia.
                                                                         CONTRIBUTION OF PERIPHERAL AND BRAIN MACRO-
                                                                         PHAGES TO GLIOMA PATHOGENESIS AND THE EFFECT OF
GLIOMA INDUCE AND EXPLOIT MICROGLIAL MT1-MMP                             SYSTEMIC CYCLOSPORIN A TREATMENT
EXPRESSION FOR TUMOR EXPANSION                                           Porycka, M.1; Gabrusiewicz, K.1; Kwiatkowska, E.2;
Markovic, DS 1,2; Chirasani, S1; Synowitz, M 1,3; Sliwa, M 4;            Kowalczyk, D.2; Kaminska, B.1
van Rooijen, N 5; Holmbeck, K6; Kiwit, J 2; Kaminska, B 4; Glass,        1Laboratory of Transcription Regulation, Department of Cell
R 1; Kettenmann, H 1                                                     Biology, Nencki Institute, Warsaw, Poland; 2Department of
Neurosurgery, Helios Klinikum, Berlin                                    Cancer Diagnostics and Immunology, Great Poland Cancer
Glioma invasion of the brain is achived by destruction of ECM            Center, Poznan, Poland;
predominantly by secretion and exrtracellular activation of matrix       Malignant gliomas are characterized by infiltration of tumour tissue
metalloproteinase 2 (MMP-2). The potent proteinase is secreted           with brain macrophages which promote tumour growth and
in zymogen, inactive form and only by interaction with distinct          invasiveness. The characteristic of immune cell infiltrating gliomas,
extracellular proteins it become activated and capable to destruct       and contribution of resident microglia and peripheral monocyte/
ECM. Crucial activator of proMMP-2 is membrane bound proteinase          macrophages is poorly defined. The aim of this study was to
membarne type one matrix metalloproteinase (MT1-MMP). We he              evaluate if peripheral macrophages migrate and contribute to
shown prevously (Markovic wt al. 2005) that the microglia in             glioma development. We generated chimeric mice with the immune
glioma surrounding is promoting glioma invasiveness by activating        system reconstituted after irradiation with hematopoietic GFP-
of MMP-2. In following study we investigated weather microglial          bone marrow cells. Then, we implanted dsRed-GL261 glioma
expression of MT1-MMP is the factor which activates MMP-2                cells into the brains of 16-week old C57BL/6 chimeric mice. Two
secreted by glioma cells. Immunohistochemistry of exhibited co-          weeks after glioma cells implantation, tumor hemispheres were
labelling of microglia with MT1-MMP. Moreover, in glioma in vivo         isolated and the abundance of the CD11b+CD45low microglia
mouse model we detected co-expression of microglia and MT1-              cells and CD11b+CD45high macrophage population was
MMP and we demonstrate that the expression of MT1-MMP is                 determined by flow cytometer. We found that peripheral GFP+
stronger in glioma surrounding than in the oposite hemisphere.           macrophages comprise above sixty percent GFP+ cells migrating
The PCR, Western Blot and MT1-MMP activity measurments in                into the tumor. Furthermore, the migration of GFP-blood-borne
cell culture experiments showed that the factor released from            macrophages into implanted dsRed-GL261 gliomas was detected
gliomas stimulates overexpression of MT1-MMP in microglia. An            using confocal microscopy. We previously demonstrated that
p38 MAPK blocker was able to block over-expression of MT1-               activation of macrophages/microglia and their promoting effects
MMP in microglia by glioma released factor. Interestingly, the TGF-      on glioma invasiveness in vitro and in vivo can be inhibited by
β stimulation of microglia mimiced effects of glioma stimulation         cyclosporin A. Therefore, we evaluated a percentage of microglial
indicating that the TGF-β is candidate for induced expression of         cells in tumour-bearing hemisphere of mice untreated or treated
MT1-MMP and by that contrinbutes in glioma invasiveness. Finally,        with cyclosporin A (10 mg/kg CsA). Drug treatment strongly
we demonstrate in glioma in vivo model that the tumor size is            reduced the number of microglial cells in tumor-bearing
smaller in MT1-MMP KO mice compared to their heterozygous                hemisphere and diminished a percentage of blood-borne
littermates.                                                             macrophages in the brain. Our findings demonstrate a significant
                                                                         infiltration of gliomas by blood-borne macrophages, which
                                                                         contribute to the pool of tumour-associated macrophages and
IN VIVO BIODISTRIBUTION AND MICROVASCULAR BINDING                        are sensitive to CsA affecting their migration and accumulation in
OF A HIGH-AFFINITY MONOCLONAL ANTIBODY                                   gliomas.
FRAGMENT F8-SIP AGAINST THE EXTRA-DOMAIN A OF
FIBRONECTIN
Parmaksiz G1, Czabanka M1, Neri D2, Vajkoczy P1.                         EXPRESSION AND FUNCTION OF THE LET7 MICRORNA
1 Department of Neurosurgery, Charite Universitätsmedizin,               DURING MOUSE CNS DEVELOPMENT
Berlin, Germany, 2 Institute of Pharmaceutical Sciences,                 Agnieszka Rybak, Heiko Fuchs, Lena Smirnova, Robert Nitsch,
Department of Chemistry and Applied Biosciences, ETH                     Gregory Wulczyn.
                                                                         Center for Anatomy, Institute of Cell Biology and
Zurich
                                                                         Neurobiology, Charite University Hospital Berlin,
Aim: The human monoclonal antibody fragment F8-SIP (small
                                                                         Phillippstrasse 12, 10115 Berlin, Germany
immunoprotein) is specific to the angiogenic marker EDA domain
                                                                         Neural development is a highly complex process that requires
of fibronectin. The aim of our study was to characterize
                                                                         precise control of gene expression. Crucial components of this
microvascular binding and biodistribution characteristics of F8-
                                                                         gene regulation machinery are small noncoding RNAs - microRNA.
SIP. Methods: SF126 cells were implanted into dorsal skin chamber
                                                                         A large number of microRNA genes are abundantly expressed in
(DSC) preparations of nude mice. Microvascular and interstitial
                                                                         the mouse nervous system but the roles of individual miRNAs in
accumulation of F8-SIP and microvascular blood flow rate were
                                                                         neural development have been not yet well characterized. In
analysed at t0 h, t2 h, t4 h and t24 h after intraarterial application



                                                                                                                                          23
Brain Tumor 2008


our study we have challanged to decipher the expression and              Glioblastomas, like other cancers, harbor small cell populations
function the let-7 microRNA family, during development of the            with the capability of sustaining tumor formation. These cells are
CNS as well as during neural stem differentiation. We have               referred to as cancer stem cells. We isolated cells expressing
found that mature form of let-7 miRNAs are not expressed in              the surface marker A2B5 from a glioblastoma and showed that
undifferentiated embryonic stem (ES) cell and related                    after grafting into nude mice, they generated dense and highly
embryocarcinoma (EC) cell lines, but are induced upon stimulation        infiltrative tumors. Then we extensively studied A2B5 + cells
of neural differentiation. To identified genes regulated by let-7,       isolated from 11 human glioblastomas. These cells display
we performed microarray profiling of mRNA after ectopic                  neurosphere-like, self-renewal, asymmetrical cell division
expression of let-7 in EC cells. A majority of the most highly           properties and have multipotency capability. Stereotactic
downregulated genes correspond to direct let-7 targets, with a           xenografts of dissociated A2B5 +-derived secondary spheres
function in early development, control of cell growth, and miRNA         revealed that as few as 1 000 cells produced a tumor. Moreover,
biogenesis. Sensor constructs confirmed let-7 sensitivity for            flow cytometry characterization of A2B5 + -derived spheres
three novel stemness genes (hic2, arid3b and mlin-41), identifying       revealed three distinct populations of cells: A2B5+/CD133+, A2B5+/
these genes as candidate targets for let-7 during embryonic              CD133- and A2B5-/CD133-, with striking proportion differences
development. Detailed analysis of mLin-41 and let-7 expression           among glioblastomas. Both A2B5+/CD133+ and A2B5+/CD133-
patterns revealed a reciprocal relationship in several pluripotent       cell fractions displayed a high proliferative index, the potential to
cell environments: ES cells, the embryonic epiblast and the male         generate spheres and produced tumors in nude mice. Finally,
germ line. We will present current data detailing expression and         we generated two GFP-cell lines that display, after serum
function of let-7/mlin-41 regulatory pair during mouse development       induction, distinct proliferative and migratory properties, in relation
and in particular the early CNS.                                         with their CD133 level of expression. Taken together, our results
                                                                         suggest that transformed A2B5+ cells are crucial for the initiation
                                                                         and maintenance of glioblastomas while CD133 expression is
DIFFERENTIATION BETWEEN MALIGNANT TRANSFORMA-                            more involved in determining the tumor’s behaviour.
TION AND TUMOUR RECURRENCE BY 68GA-BOMBESIN
AND 18F-FDG-PET IN PATIENTS WITH LOW GRADE
GLIOMAS                                                                  IDENTIFICATION OF GENES REGULATED BY MTOR IN
Seiz, M.; Dimitrakopoulou-Strauss A,; Schubert, G.A.;                    TUBEROUS SCLEROSIS AND MALIGNANT GLIOMAS
Brockmann, C.; Strauss, L.G.; Eisenhut, M.; Tüttenberg, J.               Tyburczy M.E.1; Kotulska K.2; Jozwiak S.2; Pokarowski P.3;
Universitätsmedizin Mannheim, Neurochirurgische                          Mieczkowski J.3; Kaminska B.1
Universitätsklinik, Germany                                              1 Nencki Institute of Experimental Biology, Warsaw, Poland, 2
Treatment of gliomas is multimodal. Magnetic resonance imaging           The Children’s Memorial Health Institute, Warsaw, Poland, 3
(MRI) in the posttreatment course is of limited value due to therapy-    Warsaw University, Poland
induced changes. In low-grade gliomas (LGG) malignant                    Tuberous sclerosis (TS) is a disease characterized by formation
transformation is of special interest. Our patients and methods          of benign tumors in the brain (subependymal giant cell
were as follows: In nine consecutive patients with LGG we                astrocytoma – SEGA) and neurological disorders. TS is caused
examined the role of bombesin labelled with gallium-68 (68Ga-            by mutations in TSC1 (Hamartin) or TSC2 (Tuberin) leading to
bombesin) studied with positron emission tomography (PET), in            enhancement of mTOR kinase activity and pathological alterations.
addition to fluoro-18-fluorodeoxyglucose ( 18 F-FDG) in the              mTOR is often aberrantly activated in malignant gliomas, including
differential diagnosis of tumour recurrence versus malignant             glioblastoma multiforme with frequent mutations in tumor
transformation. We used 68Ga-bombesin combined with 18F-FDG-             suppressor PTEN. mTOR activity is inhibited by Rapamycin which
PET in these patients with suspicious new contrast enhancement           is under clinical trials for TS and brain tumors. Signaling via
at the original tumour site or resection cavity in MRI. Eight patients   mTOR participates in complex genomic response but mTOR
were operated. In one patient, tumour recurrence was most                effectors in the brain are largely unknown. We sought to identify
likely as shown by the PET findings and chemotherapy was                 components of mTOR signaling network in the pathological brain.
administered. Our results have shown that in this last mentioned         Gene expression profiling was performed on four SEGAs and
patient after the follow-up period, MRI contrast enhancement             control brain samples with Affymetrix Human Genome arrays.
was definitively regressive. In the operated patients the tumour         The differences in gene expression were validated by qRT-PCR
was graded as glioblastoma multiforme (2), gliosarcoma (1) and           and studied in 5 independent cases. Identified genes encode
WHO grade III tumour (3). In two patients histological grading           proteins implicated in the nervous system development/
confirmed the PET findings without malignant transformation. In          differentiation (downregulated in TS), and proteins involved in
all of the 9 patients the combination of 68Ga-bombesin and 18F-          regulation of tumor cell proliferation (upregulated in TS). Several
FDG-PET predicted correctly malignant transformation or                  of selected genes were identified as putative mTOR effector
recurrence of the initial tumour grade which shows that 68Ga-            genes in cell cultures derived from SEGA and T98G glioblastoma
bombesin-PET can provide additional important information to             cells which exhibit high mTOR activity. Treatment with Rapamycin
detect a malignant transformation. In conclusion it is crucial for       modulated the expression of four genes in those cells
the patient to differentiate the nature of the new lesion in order to    demonstrating their association with mTOR pathway. In the
endorse an aggressive or non-aggressive treatment.                       present study several genes were identified, for the first time,
                                                                         as regulated by mTOR activity on transcriptional level in benign
                                                                         (SEGA) and malignant (T98G cell line) brain tumors.
A2B5 CELLS FROM HUMAN GLIOBLASTOMA HAVE
CANCER STEM CELL PROPERTIES
Aurélie Tchoghandjian1*, Nathalie Baeza1*, Carole Colin1,                DISSECTING CYTOKINE NETWORKS MEDIATING
Myriam Cayre2, Philippe Metellus3, Christophe Beclin2,
L’Houcine Ouafik1 and Dominique Figarella-Branger1, 4                    INTRACRANIAL TUMOR CONTROL
                                                                         Johannes vom Berg, Sergio Haller and Burkhard Becher
1Université de la Méditerranée Aix-Marseille II, Faculté de
                                                                         Institute of Experimental Immunology, Department of
Médecine la Timone, INSERM UMR911-CRO2, «
                                                                         Pathology, University Hospital of Zurich, Y44-J94,
Angiogenèse, Invasivité et Microenvironnement Tumoral », F-
                                                                         Winterthurer Strasse 190, 8057 Zurich, Switzerland
13000, Marseille, France ; 2Université de la Méditerranée
                                                                         Gliomas are the most frequently occurring brain tumors in adults.
Aix-Marseille II, Faculté de Sciences Luminy, UMR CNRS
                                                                         With its invasive and destructive growth pattern, Glioblastoma
6216 Institut de Biologie du Développement de Marseille-
                                                                         Multiforme (GBM) is the most malignant tumor among these. The
Luminy, F-13009 Marseille, France ;3Assistance Publique-
                                                                         failure of conventional treatment and the immunoprivileged status
Hôpitaux de Marseille, CHU Timone, Service de
                                                                         of the brain makes immunotherapy a promising candidate for
Neurochirurgie, Marseille, F-13000, France ; 4Assistance
                                                                         alternative treatment. To investigate the role of proinflammatory
Publique-Hôpitaux de Marseille, CHU Timone, Service
                                                                         cytokines in triggering an immune response towards GBM, we
d’Anatomie Pathologique et de Neuropathologie, Marseille, F-
                                                                         simulate human glioma in mice by using the well established
13000, France.
* These two authors have contributed equally to this study.


24
                                                                                                               Brain Tumor 2008


murine Glioma Gl261. The growth of luciferase-positive glioma         FAS SIGNALING REGULATES INVASIVENESS OF GLIOMA
cells expressing the cytokine of interest is monitored by highly      CELLS AND MMP-2 ACTIVITY VIA NFKB DEPENDENT
sensitive in-vivo bioluminescence imaging. This allows us to          MODULATION OF TIMP-2 EXPRESSION AND SECRETION
analyse the effects of cytokines on tumor growth at high spatial      Pawel Wisniewski, Aleksandra Ellert-Miklaszewska, Aneta
                                                                      Kwiatkowska, Bozena Kaminska
and temporal resolution and in a non-invasive manner. Moreover,
                                                                      Laboratory of Transcription Regulation, The Nencki Institute
we plan to unravel the enigmatic contribution of microglia/
                                                                      of Experimental Biology, Warsaw, Poland
macrohages and dendritic cells to cytokine induced glioma rejection
                                                                      Invasiveness of malignant gliomas strongly contributes to the
at various stages of tumor growth via cellular depletion using
                                                                      failure of current treatments and poor survival of patients. Tumor
inducible genetic models. Together with FACS analysis and
                                                                      infiltrating brain microglia/macrophages support glioma growth
histology we will acquire a highly detailed picture of the complex
                                                                      and invasion. Mechanisms underlying microglia-dependent
interplay of various cellular effectors and signal molecules that
                                                                      influence on glioma invasion are poorly understood. Microglia
lead to intracranial tumor control.
                                                                      are the main source of cytokine – FasL in the brain, which is able
                                                                      to induce apoptosis in Fas bearing cells, but may also play
                                                                      different, non-apoptotic roles in many cell types, including gliomas.
BRDU LABELING PATTERN OF THE ROSTRAL MIGRATORY
                                                                      To eveluate an involvement of Fas signaling in the regulation of
STREAM IN NORMAL CANINE AND FELINE BRAINS
Watson, D.; Malik, S.; Vite, C.; Haskins, M.; Van Winkle, T.          glioma invasiveness, we developed a recombinant FasL
Department of Neurosurgery and School of Veterinary                   Interfering Protein (FIP) which has been tested on C6 glioma
Medicine, University of Pennsylvania, Philadelphia, PA, USA           cells expressing both Fas receptor and its ligand. FIP
In the rodent and human brain, neural progenitor cells migrate in     administration did not affect a cell viability but impaired migration
the rostral migratory stream extending from the subventricular        (in scratch assay) and invasiveness of glioma cells (in Matrigel
zone (SVZ) of the lateral ventricle towards the olfactory bulb.       assay). Blocking of Fas signaling reduced MMP-2 activity in glioma
We are characterizing the pattern of proliferative cells in the       cells. To study signaling events affected by FIP treatment, we
normal canine and feline brain using 5-bromo-2-deoxyuridine           evaluated FIP effects on MAPK signaling and activities of AP-1
(BrdU) labeling. BrdU was administered intravenously to dogs          and NFkB transcription factors, commonly associated
(n=4, 25 or 75 mg/kg) or cats (n=4, 25 mg/kg) every 24hr for 4        withregulation of invasion. Interference with Fas signaling
days. Dogs were perfused at 1 or 5 days, and cats were                reduced JNK activity, AP-1 and NFkB-driven transcription. FIP
perfused at 1 or 10 days, following the final infusion. Two dogs      treatment did not affect mmp-2 and mmp-14 expression but
and one cat received a single dose of 75 mg/kg and were               significantly attenuated timp-2 expression and TIMP-2 amount in
perfused 6 hr later. Frozen sagittal sections were stained with       the culture medium. Studies with pharmacological inhibitors of c-
Nissl, H&E, or anti-BrdU antibody. We identified a prominent,         Jun N-Terminal Kinase (SP600125) and NFkB (BAY 11-7082)
dense, continuous track of Nissl-stained cells, beginning at the      signaling pathways revealed that timp-2 expression is regulated
base of the anterior horn of the lateral ventricle, curving around    by NFkB transcription factor, whereas its secretion is regulated
the rostral portion of the head of the caudate nucleus and            by both NFkB and JNK pathways. Our findings show that Fas
continuing ventrally to the olfactory peduncle. BrdU-                 signaling regulates invasiveness of glioma cells via regulation of
immunoreactive cells were present in the same track. The BrdU-        MMP-2 activation, most likely by controlling TIMP-2 expression
labeled nuclei were single or closely apposed small groups and        and secretion.
many were located at the boundary between the caudate nucleus
and the rostral white matter. A similar pattern of staining was
found in dogs receiving one dose of BrdU, indicating cell division
continuously along the track. The distribution of these cells
appears analogous to the rostral migratory stream found in other
species. We will present our ongoing studies to describe the
phenotype of these cells and the anatomical route and dimensions
of the track. Grant Support: Penn Neurosurgery and NIH RR02512.




                                                                                                                                       25
Brain Tumor 2008




26
                                                                                                                             Brain Tumor 2008


                                                           Address List
Abdalla, Dr. Yasser                                   Bjerkvig, Prof. Dr. Rolf                          Clément, Dr. Virginie
Krankenhaus                                           University of Bergen                              Geneva University Hospitals and University of
Neurochirurgie                                        NorLux Neuro-Oncology, Dept. of Biomedicine       Geneva
Hauptstraße                                           Jonas Lies vei 91                                 Centre Romand de Neurochirurgie
26452 Sande ( Germany)                                N-5009 Bergen (Norway)                            Micheli-du-Crest, 24
Phone: 04422-801450                                   Email: rolf.bjerkvig@pki.uib.no                   1211 Geneva (Switzerland)
Email: y.abdalla@sanderbusch.de                                                                         Phone: +41 22 3723425
                                                      Bogdahn, Prof. Dr. Ulrich                         Email: virginie.clement@medecine.unige.ch
Abounader, Professor Roger                            University of Regensburg
University of Virginia                                Neurology                                         Colin, Dr. Carole
Neurology                                             Universitaetsstrasse 84                           Université de la Méditerranée - Faculté de
PO Box 800168                                         93053 Regensburg (Germany)                        Médecine Timone
22903 Charlottesville, VA (USA)                       Phone: +49 9419413001                             INSERM U911 - CRO2
Phone: 434-982-6634                                   Email: ulrich.bogdahn@medbo.de                    27, Bd Jean Moulin
Email: ra6u@virginia.edu                                                                                13005 Marseille (France)
                                                      Bothmann, Dr. med. Matthias                       Phone: +334 91 32 44 43
Albrecht, Dr. Valerie                                 Kantonsspital Winterthur                          Email: carole.colin@univmed.fr
Ludwig-Maximilians-University                         Neurochirurgie
Neurosurgical Clinic Grosshadern                      Brauerstrasse 15                                  Corno, Dr. Daniela
Marchioninistr. 15                                    8401 Winterthur (Switzerland)                     San Raffaele Scientific Institute
81377 München                                         Email: matthias.bothmann@ksw.ch                   Stem Cell Research Institute
Email: valerie.albrecht@med.uni-muenchen.de                                                             Via Olgettina 58
                                                      Bottino, Prof Cristina                            20132 Milan (Italy)
Auf, Dr. Gregor                                       University of Genova                              Phone: +39-02-26434630
Université Bordeaux 1                                 Dipartimento di Medicina Sperimentale and         Email: corno.daniela@hsr.it
INSERM U920                                           Istituto G. Gaslini
43, Avenue des Facultés                               L.go G. Gaslini 5                                 Cuevas, Elisa
33405 Talence (France)                                16148 Genova (Italy)                              Charité-Universitätsmedizin
Email: gregor.auf@charite.de                          Email: Cristina.Bottino@unige.it                  Institut of Cell and Neurobiology, Center for
                                                                                                        Anatomy
Baeza, Dr Nathalie                                    Bräter, Kathleen                                  Philippstr. 12
Université de la Méditerranée - Faculté de Médecine   Paracelsus Klinik Zwickau                         10115 Berlin (Germany)
Timone                                                Neurochirurgie                                    Phone: 030450528245
INSERM U911 - CRO2                                    Werdauer Straße 68                                Email: elisa.cuevas@charite.de
27, Bd Jean Moulin                                    08008 Zwickau
13005 Marseille (France)                              Email: kathleen_braeter@gmx.de                    Czabanka, Dr. Marcus
Phone: +334 91 32 44 43                                                                                 Universitätsmedizin Charite
Email: nathalie.baeza@univmed.fr                      Brandner, Prof Sebastian                          Dept. of Neurosurgery
                                                      UCL Institute of Neurology                        Augustenburger Platz1
Barbus, Sebastian                                     Divsion of Neuropathology                         13353 Berlin (Germany)
DKFZ                                                  Queen Square                                      Phone: 0049 30 450 66029
Molecular Genetics                                    WC1N 3BG London (UK)                              Email: marcus.czabanka@charite.de
INF 580                                               Phone: 0044 20 7676 2188
69120 Heidelberg                                      Email: s.brandner@ion.ucl.ac.uk                   Cziomer, Jessica
Email: s.barbus@dkfz.de                                                                                 Bundeswehrzentralkrankenhaus Koblenz
                                                      Braun, Christian                                  Neurosurgery
Barciszewska, MD, MSc, MA Anna-Maria                  Universität Tübingen                              Rübenacherstraße 172
Karol Marcinkowski University of Medical Sciences     Allgemeine Neurologie                             56072 Koblenz ()
Chair and Clinic of Neurosurgery and                  Hoppe-Seyler-Straße 3                             Phone: 0261-3027963
Neurotraumatology                                     72076 Tübingen ( Germany)                         Email: jes.c@debitel.net
Przybyszewskiego 49                                   Phone: 07071/2982058
60-355 Poznan (Poland)                                Email: christian.braun@medizin.uni-tuebingen.de   Diestel, Dr. Antje
Phone: + 48 61 8691422                                                                                  Charite
Email: abarcisz@man.poznan.pl                         Braunsdorf, Dr. Werner EK.                        Paediatric Cardiology
                                                      Klinikum Magdeburg gGmbH                          Augustenburger Platz 1
Barrantes Freer, Alonso                               Neurosurgery                                      13353 Berlin (Germany)
Max Planck Institut für Experimentelle Medizin        Birkenallee 34                                    Phone: 030450559886
Molecular Biology of Neuronal Signals                 39130 Magdeburg (Germany)                         Email: antje.diestel@charite.de
Herman Rein Strasse 3.                                Phone: +49-391-7914701
37075 Göttingen ( Germany)                            Email: braunsdorf@klinikum-magdeburg.de           Dirks, Dr. Peter B.
Phone: 0 176 646 22217                                                                                  The Arthur and Sonia Labatt Tumor Res. Ctr.
Email: barrantes@em.mpg.de                            Brüggemann, Dr. Chantal                           The Hospital of Sick Children
                                                      Max-Planck-Institute for neurological research    555 University Avenue
Bernreuther, Dr. Christian                            in-vivo-NMR-Laboratory                            Toronto, Ontario Canada
University Medical Center Hamburg-Eppendorf           Gleueler Str. 50                                  Email: peter.dirks@sickkids.ca
Institute of Neuropathology                           50931 Köln
Martinistrasse 52                                     Email: brueggemann@nf.mpg.de                      Dondero, Dr Alessandra
20246 Hamburg                                                                                           University of Genova
Email: c.bernreuther@uke.uni-hamburg.de               Burkhardt, Dr. Jan-Karl                           Department of Experimental Medicine
                                                      University Hospital Zürich                        Via Marsano 10
Bessette, PhD Barbara                                 Dep. of Neurosurgery                              16132 Genova
Faculté de Médecine                                   Frauenklinikstr.10, Nord1, D                      Email: alessandra.dondero@unige.it
EA3842 Homéostasie cellulaire et pathologies          8091 Zürich (Schweiz)
2 rue du Docteur Marcland                             Email: Jan-Karl.Burkhardt@usz.ch                  El Demery, Dr Mounira
87125 Limoges (France)                                                                                  CHU Limoges
Email: barbara.bessette@laposte.net                   Castriconi, Dr Roberta                            medical oncology
                                                      Via Marsano 10                                    2 avenue M.L.King
Bexell, Daniel                                        16155 Genova (Italy)                              87000 Limoges (France)
Lund Stem Cell Center, BMC B10                        Email: Roberta.Castriconi@unige.it                Phone: 003355056100
Klinikgatan 26                                                                                          Email: mounira.eldemery@chu-limoges.fr
221 84 Lund (Sweden)                                  Cheray, Miss Mathilde
Email: daniel.bexell@med.lu.se                        Université de Limoges                             El Hallani, Soufiane
                                                      Lab. Neuro-immunologie, Faculté de Médecine       Salpetriere Hospital
Bick-Sander, Dr. med. Anika                           2, rue du Dr Marcland                             MAZARIN Neuro-oncology
Charite, Campus VIrchow Klinikum                      87025 Limoges (France)                            17, rue de Sèvres
Department for Neurology                              Phone: +33 5 55 43 58 65                          75006 Paris (France)
Augustenburger Platz 1                                Email: mathilde.cheray@etu.unilim.fr              Phone: +33 06 30 46 55
13353 Berlin Berlin (Germany)                                                                           Email: elhallanisoufiane@yahoo.fr
Phone: 030 450 660235                                 Cholewa, Jadwiga
Email: anika.bick-sander@charite.de                   MDC                                               Elfituri, Dr. Nemrish
                                                      Developmental Neurobiology                        Tawam Krankenhaus
Birkenmeier, Prof. Gerd                               Robert-Rössle-Str. 10                             Neurochirurgie
University of Leipzig                                 13125 Berlin                                      Almuqam
Institute of Biochemistry                             Email: jadwiga.cholewa@mdc-berlin.de              15258 Al ain city (United Arab Emirates)
Johannisallee 30                                                                                        Phone: 00971507639020
04103 Leipzig (Germany)                               Ciechomska, Dr Iwona                              Email: nemrish@yahoo.de
Phone: 0341 9722132                                   Institute of Experimental Biology
Email: Gerd.Birkenmeier@medizin.uni-leipzig.de        Department of Cell Biology
                                                      3 Pasteur
                                                      02-093 Warsaw (Poland)
                                                      Email: jcjech@nencki.gov.pl


                                                                                                                                                        27
Brain Tumor 2008


Eltabeb, Dr. Fatma                                   Hahnen, Dr. Eric                                    Kettenmann, Prof. Dr. Helmut
Tawam Krankenhaus                                    Institute of Human Genetics                         MDC
Gynacology                                           Kerpener Str. 34                                    Cellular Neurosciences
Al muqam                                             50931 Cologne (Germany)                             Robert-Rössle-Str. 10
15258 Al ain (United Arab Emirates)                  Phone: +49 221 478-86464                            13125 Berlin (Germany)
Phone: 00971508313121                                Email: hahnen@rocketmail.com                        Phone: 030 9406 3325
Email: sohear65@yahoo.com                                                                                Email: kettenmann@mdc-berlin.de
                                                     Hattermann, Kirsten
Endres, Prof. Dr. Matthias                           Universität Kiel                                    Kirsch, Priv.-Doz. D Dr Matthias
Charité - Universitätsmedizin Berlin, Campus Mitte   Anatomie                                            Carl Gustav Carus Universitätsklinikum Dresden
Klinik nund Poliklinik für Neurologie                Olshausenstr. 40                                    Neurochirurgie
Schumannstr. 20/21                                   24098 Kiel (Germany)                                Fetscherstrasse 74
10117 Berlin (Germany)                               Phone: +49 431 880 2460                             01307 Dresden ()
Phone: +49 30 450 560102                             Email: k.hattermann@anat.uni-kiel.de                Phone: 0351 458 4735
Email: matthias.endres@charite.de                                                                        Email: matthias.kirsch@uniklinikum-dresden.de
                                                     Hau, PD Dr. med. Peter
Eyupoglu, Dr. med. Ilker                             University of Regensburg                            Kiwit, Prof. Dr. Jürgen
University of Erlangen-Nuremberg                     Neurology                                           Helios Klinikum Buch
Department of Neurosurgery                           Universitätsstrasse 84                              Neurochirurgische Klinik
Schwabachanlage 6                                    93053 Regensburg (Germany)                          Schwanebecker Chaussee 50
91054 Erlangen (Germany)                             Phone: 0941 941-8083                                13125 Berlin ()
Phone: +49 9131 85 44756                             Email: peter.hau@medbo.de                           Email: juergen.kiwit@berlin.helios-kliniken.de
Email: ilker.eyupoglu@uk-erlangen.de
                                                     Heine, Angela                                       Kleindienst, PD Dr. Andrea
Feltz, Dr. Rüdiger                                   FU Berlin                                           Dept. of Neurosurgery
HELIOS-Klinikum Erfurt                               Psychologie                                         Mariahilfbergweg 6
Neurosurgery                                         Habelschwerdter Allee 45                            92224 Amberg
Nordhäuser Str.74                                    14195 Berlin                                        Email: andrea.kleindienst@uk-erlangen.de
99084 ERFURT ()                                      Email: aheine@zedat.fu-berlin.de
Email: ruediger.feltz@helios-kliniken.de                                                                 Kruttwig, Klaus
                                                     Heinicke, Dr. Dirk                                  Max-Planck-Institute for neurological research
Finocchiaro, Dr . Gaetano                            Fiebigweg 19                                        in-vivo-NMR-Laboratory
Fondazione IRCCS Istituto Neurologico C. Besta       01731 Kreischa                                      Gleueler Str. 50
Experimental Neuro-Oncology                          Email: kud.heinicke@gmx.de                          50931 Köln (Germany)
Via Celoria 11                                                                                           Phone: 02214726335
20133 Milano (Italy)                                 Heppner, Prof. Dr. Frank                            Email: kruttwig@nf.mpg.de
Phone: 02 2394 2285                                  Medizinische Fakultät der HU
Email: gaetano.finocchiaro@gmail.com                 Institut für Neuropathologie                        Ku, Min-Chi
                                                     Augustenburger Platz 1                              MDC
Fiss, Dr. Ingo                                       13353 Berlin (Germany)                              Robert-Rossle str. 10
Charite - Universitätsmedizin Berlin                 Phone: +49 30 450 536043                            13125 Berlin (Germany)
Neurosurgery                                         Email: frank.heppner@charite.de                     Email: min-chi.ku@mdc-berlin.de
Hindenburgdamm 30
12169 Berlin ( Germany)                              Hoene, Victoria                                     Lamszus, Dr. Katrin
Phone: 8445-2610                                     Charité Universitätsmedizin Berlin, CVK             University Medical Center Hamburg-Eppendorf
Email: ingo.fiss@charite.de                          Neonatology                                         Laboratory for Brain Tumor Biology, Dept. of
                                                     Augustenburger Platz 1, Forschungshaus (Forum       Neurosurgery
Fuchs, PhD sudent Heiko                              13353 Berlin                                        Martinistrasse 52
Institute for Cell biology and Neurobiology          Email: victoria.hoene@charite.de                    20246 Hamburg (Germany)
Center for Anatomy                                                                                       Phone: 040 42803-5577 or
Phillippstr. 12                                      Höpner, Sabine                                      Email: lamszus@uke.uni-hamburg.de
10115 Berlin (Germany)                               MDC-Berlin
Email: heiko.fuchs@charite.de                        Robert-Rössle-Str.10                                Landfried, Britta
                                                     13125 Berlin ( Germany)                             Abteilung Neuropathologie
Gabrusiewicz, Konrad                                 Email: shoepner@mdc-berlin.de                       Institut für Pathologie, Jena
The Nencki Institute of Experimental Biology                                                             Unterm Markt 4
Department of Cell Biology                           Holland, Dipl.-Biol. Heidrun                        07743 Jena (Germany)
3 Pasteur Street                                     Translational Centre for Regenerative Medicine      Phone: 03641-376444; 017
02-093 Warsaw (Poland)                               University of Leipzig                               Email: Britta.Landfried@uni-jena.de
Email: k.gabrusiewicz@nencki.gov.pl                  Johannisallee 30
                                                     04103 Leipzig (Germany)                             Leutz, Prof. Dr. Achim
Galli, Dr. Rossella                                  Phone: 0341-9725484                                 MDC
San Raffaele Scientific Institute                    Email: Heidrun.Holland@medizin.uni-leipzig.de       Tumorgenese und Zelldifferenzierung
Stem Cell Research Institute                                                                             Robert-Rössle-Str. 10
Via Olgettina 58                                     Holtkamp, Nikola                                    13125 Berlin (Germany)
20132 Milan (Italy)                                  Charité-Universitätsmedizin Berlin                  Phone: +49 30 94063735
Phone: +39-02-26434626                               Institut für Neuropathologie                        Email: aleutz@mdc-berlin.de
Email: galli.rossella@hsr.it                         Charitéplatz 1
                                                     10117 Berlin                                        Liesenberg, Franziska
Gaunitz, Priv.-Doz. Dr. Frank                        Email: nikola.holtkamp@charite.de                   Neuropathologie
University of Leipzig                                                                                    Uniklinik Düsseldorf
Institut of Biochemistry                             Hutschenreuther, Antje                              Moorenstr. 5
Johannisallee 30                                     Biochemistry                                        40225 Düsseldorf (Germany)
04103 Leipzig (Germany)                              Medical Faculty                                     Phone: 0211 8118652
Phone: +49 341 9722153                               Johannisallee 30                                    Email: franziska.liesenberg@t-online.de
Email: fgau@medizin.uni-leipzig.de                   04103 Leipzig (Germany)
                                                     Email: Antje.Hutschenreuther@medizin.uni-           Lipko, Dr. Maciej
Glass, Dr. Rainer                                    leipzig.de                                          NENCKI Institute of Experimental Biology
MDC                                                                                                      Cell Biology
Cellular Neurosciences                               Inam, Dr.med. Kassir                                Pasteur 3
Robert-Rössle-Str. 10                                Ev.Bathildiskrankenhaus Bad Pyrmont gGmbH           02-093 Warsaw (Poland)
13125 Berlin (Germany)                               Neurochirurgische Abteilung                         Email: mlipko@nencki.gov.pl
Phone: +49 30 9506 3260                              Maulbeerallee 4
Email: rainer.glass@mdc-berlin.de                    31812 Bad Pyrmont (Germany)                         Löhr, Dr Mario
                                                     Phone: 05281 992002                                 University Hospital
Haag, Daniel                                         Email: inam.kassir.@bathildis.de                    Neurosurgery
Dkfz                                                                                                     Kerpener Strasse 62
Molecular Genetics                                   Kälin, Dr. Roland                                   50937 Köln (Germany)
Im Neuenheimer Feld 580                              Charite Berlin                                      Phone: 0221-478 4576 / 4
69120 Heidelberg (Germany)                           Neuropathologie                                     Email: Mario.Loehr@uk-koeln.de
Phone: 06221424594                                   Fehrbellinerstr. 31
Email: d.haag@dkfz.de                                10117 Berlin                                        Markovic, MD/PhD Darko
                                                     Email: roland.kaelin@charite.de                     Helios Klinikum
                                                                                                         Neurosurgery
                                                     Kahlert, Ulf                                        Schwanebecker Chaussee 50
                                                     University Hospital Freiburg                        13125 Berlin (Germany)
                                                     Stereotactic Neurosurgery                           Phone: +4930940114386
                                                     Breisacher Straße 64                                Email: darko.markovic@helios-kliniken.de
                                                     79106 Freiburg
                                                     Email: ulf-dietrich.kahlert@uniklinik-freiburg.de   Martin-Villalba, PD Dr. Ana
                                                                                                         INF 581
                                                                                                         69120 Heidelberg
                                                                                                         Email: a.martin-villalba@dkfz.de



28
                                                                                                                        Brain Tumor 2008


Mawrin, Prof. Dr. Christian                     Roettger, Dr. Ernst                                   Teßmann, Grietje
Otto-von-Guericke-Universität Magdeburg         Krankenhaus Ludmillenstift                            MDC
Neuropathologie                                 Neurochirurgie                                        Cellular Neuroscience
Leipziger Strasse 44                            Landwehr 32                                           Robert-Rössle-Straße 10
39120 Magdeburg                                 49716 Meppen ( Germany)                               13092 Berlin ( Germany)
Email: christian.mawrin@med.ovgu.de             Phone: 05931 85577                                    Phone: 030-9406-3503
                                                Email: maria.roettger@ewetel.net                      Email: grietje.tessmann@mdc-berlin.de
Mazzoleni, Dr. Stefania
San Raffaele Scientific Institute               Rothe, Dr. med. Ronny M.                              Tews, Dr. Björn
Stem Cell Research Institute                    Hauptstr. 33                                          Brain Research Institute - ETH/University Zürich
Via Olgettina 58                                13158 Berlin                                          Winterthurerstr. 190
20132 Milan (Italy)                             Email: romaro12@aol.com                               CH-8057 Zürich (Switzerland)
Phone: +39-02-26434630                                                                                Phone: +41 44 63 53262
Email: mazzoleni.stefania@hsr.it                Rybak, PhD sudent Agnieszka                           Email: tews@hifo.uzh.ch
                                                Institute for Cell biology and Neurobiology
Momma, Dr. Stefan                               Center for Anatomy                                    Tödt, Grischa
University of Frankfurt Medical School          Phillippstr. 12                                       German Cancer Research Center
Neurological Institute (Edinger Institute)      10115 Berlin (Germany)                                Molecular Genetics
Heinrich-Hoffmann-Str.7                         Email: agnieszkarybak@yahoo.com                       Gleiwitzer Str. 19
60528 Frankfurt am Main                                                                               69124 Heidelberg
Email: stefan.momma@kgu.de                      Schmidt, Natalie                                      Email: g.toedt@dkfz.de
                                                Neuropathologie
Nakamura, PD Dr. med. Makoto                    Uniklinik Düsseldorf                                  Tröller, Dipl.-Ing. Silke
Hannover Medical University                     Moorenstr. 5                                          DHZB
Dept. of Neurosurgery                           40225 Düsseldorf (Germany)                            Kinderkardiologie
Carl-Neuberg Str. 1                             Phone: 0211 8118652                                   Augustenburger Platz 1
30625 Hannover (Germany)                        Email: natalie.schmidt@med.uni-duesseldorf.de         13353 Berlin ( Germany)
Phone: +49-511-5323770                                                                                Phone: 030-45932868
Email: nakamura.makoto@mh-hannover.de           Schmitt, Ilka                                         Email: silke.troeller@charite.de
                                                University Hospital Eppendorf, Hamburg
Oerlecke, Ilka                                  Martinistraße 52                                      Tyburczy, MSc Magdalena
Institute of Biochemistry                       20246 Hamburg (Germany)                               Nencki Institute of Experimental Biology
Medical Faculty                                 Phone: +49 40 23518813                                Department of Cell Biology
Johannisallee 30                                Email: ilkaschmitt@gmx.de                             Pasteur 3
01403 Leipzig                                                                                         02-093 Warsaw (Poland)
Email: ilka.oerlecke@medizin.uni-leipzig.de     Seiz, Dr. Marcel                                      Email: m.tyburczy@nencki.gov.pl
                                                Neurochirurgische Universitätsklinik
Pagenstecher, Prof. Dr. Axel                    Universitätsmedizin Mannheim                          Vajkoczy, Prof. Dr. med. Peter
University of Marburg                           Theodor-Kutzer-Ufer 1-3                               Charité - Campus Virchow-Klinikum
Neuropathology                                  68167 Mannheim (Germany)                              Klinik und Poliklinik für Neurochirurgie
Baldingerstr.                                   Email: marcel.seiz@nch.ma.uni-heidelberg.de           Augustenburger Platz 1
35041 Marburg                                                                                         13353 Berlin
Email: pagenste@med.uni-marburg.de              Seyffarth, Anne                                       Email: peter.vajkoczy@charite.de
                                                University Leipzig
Parmaksiz, Güliz                                Biochemistry                                          van Lohuizen, Prof. Dr. Maarten
Charite Universitaetsmedizin, Berlin, Germany   Hospitalstr. 2                                        The Netherlands Cancer Institute
Department of Neurosurgery                      04600 Altenburg (Germany)                             Divison of Molecular Genetics
Wöhlertstr 6                                    Email: anneseyffarth@web.de                           P1.015 Plesmanlaan 121, postbus 90203, 1006
10115 Berlin                                                                                          B
Email: gueliz.parmaksiz@charite.de              Söling, PD Dr. Ariane                                 1066 CX Amsterdam (The Netherlands)
                                                University of Göttingen                               Email: m.v.lohuizen@nki.nl
Peterziel, Dr. Heike                            Dept. of Pediatrics 1
DKFZ Heidelberg                                 Robert-Koch-Str. 40                                   vom Berg, Johannes
A100 - Signal Transduction and Growth Control   37075 Göttingen (Germany)                             University Hospital of Zurich
Im Neuenheimer Feld 280                         Email: asoeling@med.uni-goettingen.de                 Institute of Experimental Immunology, Department
69120 Heidelberg (Germany)                                                                            of Pathology
Phone: +49-6221-424501                          Stock, Kristin                                        Y44-J94, Winterthurer Strasse 190
Email: h.peterziel@dkfz-heidelberg.de           Max Delbrueck Center for Molecular Medicine           8057 Zurich (Switzerland)
                                                Robert-Roessle-Str. 10                                Phone: +41 44 635 3707
Phillips, Prof. Dr. Heidi                       13125 Berlin                                          Email: johannes.vomberg@neuroimm.uzh.ch
Genentech &amp; Unversity of California San     Email: kristin.stock@mdc-berlin.de
Francisco                                                                                             Waha, Dr. Andreas
Tumor Biology and Angiogenesis                  Stricker, Dr. Stefan                                  University of Bonn
1 DNA Way                                       University of Cambridge                               Neuropathology
            ,
94080 SSF CA (USA)                              CSCR Wellcome Trust Centre for Stem Cell Research     Sigmund-Freud-Strasse 25
Phone: + 1 650 225-2657                         Tennis Court Road                                     53105 Bonn
Email: hsp@gene.com                             CB2 1QR Cambridge (UK)                                Email: awaha@uni-bonn.de
                                                Phone: 01223 760282
Plate, Prof. Dr. Karlheinz                      Email: shs35@cscr.cam.ac.uk                           Waha, Dr. Anke
Frankfurt University Medical School                                                                   University of Bonn
Neurologisches Institut                         Synowitz, Dr. Michael                                 Neuropathology
Heinrich Hoffmann Strasse 7                     Charité - Universitätsmedizin Berlin                  Sigmund-Freud-Strasse 25
60528 Frankfurt/Main                            CVK, Klinik für Neurochirurgie                        53105 Bonn
Phone: +4969-63016042                           Augustenburger Platz 1                                Email: awaha1@uni-bonn.de
Email: karl-heinz.plate@kgu.de                  13353 Berlin
                                                Email: michael.synowitz@charite.de                    Watson, Dr. Deborah
Pollard, Dr Steven                                                                                    University of Pennsylvania
University of Cambridge                         Szymanski, Caroline                                   Neurosurgery
Wellcome Trust Centre for Stem Cell Research    Charité Berlin                                        371A Stemmler Hall/ 36th and Hamilton Walk
Tennis Court Road                               Master Programm Medical Neurosciences                 19146 Philadelphia, PA (USA)
CB2 1QR Cambridge (UK)                          Christburger Str. 21                                  Phone: 215-901-5289
Phone: 044 1223 760281                          10405 Berlin (Germany)                                Email: djw3@mail.med.upenn.edu
Email: smp54@cam.ac.uk                          Email: caroline.szymanski@gmx.de
                                                                                                      Wisniewski, Pawel
Porycka, Malgorzata                             Tabatabai, Dr. Ghazaleh                               The Nencki Institute of Experimental Biology
The Nencki Institute of Experimental Biology    University of Tübingen                                Department of Cell Biology
Department of Cell Biology                      Department of General Neurology                       3 Pasteur st.
3 Pasteur Street                                Hoppe-Seyler Strasse 3                                02-093 Warsaw (Poland)
02-093 Warsaw (Poland)                          72076 Tübingen (Germany)                              Email: p.wisniewski@nencki.gov.pl
Email: m.porycka@nencki.gov.pl                  Phone: 07071-298-7615
                                                Email: ghazaleh.tabatabai@uni-tuebingen.de            Wölfer, Dr. med. Johannes
Riemenschneider, Dr. Markus J.                                                                        Universitätsklinik Münster
Heinrich-Heine-University Düsseldorf            Tchoghandjian, Aurélie                                Klinik und Poliklinik für Neurochirurgie
Neuropathology                                  Université de la Méditerranée - Faculté de Médecine   Albert-Schweitzer-Straße 33
Moorenstr. 5                                    Timone                                                48149 Münster ( Germany)
40225 Düsseldorf (Germany)                      INSERM U911 - CRO2                                    Phone: +49-251-8347496
Phone: 0211-8118663                             27, Bd Jean Moulin                                    Email: woelfer@uni-muenster.de
Email: majori@gmx.net                           13005 Marseille (France)
                                                Phone: +334 91 32 44 43
                                                Email: aurelie.tchoghandjian@univmed.fr




                                                                                                                                                     29
Brain Tumor 2008


Wojtas, Dr. Kordian
Klinikum Kassel
Neurosurgery
Lange Strasse 91
34131 Kassel (Germany)
Email: kordianwojtas@web.de

Wolter, Dr. Marietta
Institut für Neuropathologie
Heinrich-Heine-Universität
Moorenstrasse 5
40225 Düsseldorf ( Germany)
Phone: 0211-811 8652
Email: wolter@med.uni-duesseldorf.de
Wulczyn, Dr. F Gregory
Charité Universitätsmedizin Berlin
Institute for Cell and Neurobiology
Phillippstrasse 12
10115 Berlin
Email: gregory.wulczyn@charite.de

Zaspel, Dr. med. Nancy
Paracelsus Klinik Zwickau
Neurochirurgie
Werdauer Straße 68
08008 Zwickau
Email: Nancy.Zaspel@gmx.de

Zille, Marietta
Koloniestraße 38
13359 Berlin (Germany)
Email: marietta.zille@charite.de

Zipp, Frauke Zipp
Wiss. Direktorin der Cecilie-Vogt-Klinik für
Neurologie im HKBB
Charité - Universitätsmedizin Berlin und Max-
Delbrück-Centrum
Schwanebecker Chaussee 50
13125 Berlin
Phone: +49 30 9401 54250
Email frauke.zipp@charite.de




30
        Brain Tumor 2008



Notes




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Brain Tumor 2008




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