bluuebunny/crossref_metadata_2025_split huggingface dataset:
SPDR-09 CHANGES IN CELL CYCLE-RELATED GENE EXPRESSIONS OF GLIOBLASTOMAS BEFORE AND IMMEDIATELY AFTER CHEMO-RADIATION THERAPY
Authors: Iuchi Toshihiko , Hara Ryusuke , Kageyama Hajime , Sugiyama Takahiro , Togasaki Gentaro , Higuchi Akio , Hosono Junji , Setoguchi Taiki , Hasegawa Yuzo , Sakaida Tsukasa , Itami Makiko 
Published: December 2019

Abstract

PURPOSE/OBJECTIVE
The molecular responses of glioblastomas (GBMs) to hypofractionated IMRT/TMZ were investigated to elucidate the molecular targets included in the resistance of these tumors to chemo-radiation therapy.

MATERIALS /METHODS
Phase I study of neo-adjuvant IMRT (72Gy/12Fx.)/TMZ for the treatment of patients with GBMs had been performed previously in our institution. In this trial, stereotactic biopsy of the tumor to confirm the pathological diagnosis prior to treatment was required, and tumor removal was scheduled within 10 days after completion of IMRT/TMZ. Therefore, both the tumor samples before and immediately after IMRT/TMZ were available. By comparing the gene expression profiles before and after IMRT/TMZ using the total mRNA sequencing (RNAseq) analysis, molecular responses of GBMs against IMRT/TMZ were investigated. More than two-fold change of expression levels was defined as significant.

RESULTS
Tumor sample sets from five patients with GBMs were investigated. Among the 17,532 genes evaluated, 35 genes were found to show significant changes in gene expression in all cases, and 450 genes in more than half of the cases. Among the DNA repair related genes, DDB2 was the only gene that showed significant up-regulation in all cases. On the other hand, among the cell cycle checkpoint related genes, gene expressions of CKD1/CCNB were decreased in all cases. Although the expression of TP53 was not changed, the expressions of CDKN1A/GADD45/Reprimo/SFN were also reduced. Moreover, although the expression change of CHK1 was not found, the expressions of CDC25/PLK1/AURKA were decreased in more than half of the cases. From these results, it was considered that GBM arrested the cell cycle at the G2/M checkpoint without regulation of TP53 or CHK1 after IMRT/TMZ.

CONCLUSIONS
Our results suggested that cell cycle arrest in G2/M plays a significant role in survival of GBM cells after IMRT/TMZ.
ET-04 MOLECULAR TARGETED THERAPY AGAINST (PRO)RENIN RECEPTOR FOR GLIOBLASTOMA
Authors: Fujimori Takeshi , Ogawa Daisuke , Suzuki Kenta , Kochi Masaaki , Shibayama Yuki , Okada Masaki , Miyake Keisuke , Nishiyama Akira , Tamiya Takashi 
Published: December 2019

Abstract

INTRODUCTION
(Pro)renin receptor(PRR) is part of the Wnt receptor complex. Wnt/β-catenin signaling pathway (Wnt signaling) plays important role in pathogenesis and self-renewal of glioblastoma (GBM), or differentiation of glioma stem cell. We previously reported that PRR activate Wnt signaling, PRR expression correlated with malignancy of glioma, and treatment with PRR siRNA reduced the proliferative capacity. This time, we have developed monoclonal antibodies against PRR and examined their effects in GBM.

MATERIAL AND METHODS
We used GBM cell line (U251MG and U87MG) and primary human glioma stem cell line (MGG23). Glioma stem-like cells were cultured and isolated by neurosphere method from U251MG and U87MG. PRR antibody was made targeting the extracellular domain of the PRR with rat lymph node method. WST-1 assay or MTT assay were performed to determine the cell proliferation. Apoptosis was examined by FITC labeled annexin V and propidium iodide with flow cytometry. We analyzed molecules of Wnt signaling and stem cell markers with qRT-PCR.

RESULTS
We observed that PRR antibody significantly reduced cell proliferation, decreased sphere formation. Antibody suppressed cell adherent in stem-like cell. Flow cytometry showed that antibody induced apoptosis. Antibody inhibited Wnt signaling and stem cell markers.

CONCLUSIONS
PRR antibody reduced cell proliferation and induced apoptosis through Wnt signaling. PRR antibody also suppressed stemness. Our results demonstrated that PRR was a potential target for future glioma therapy.
ET-09 ACQUIRED MALIGNANT BEHAVIORS OF NPE6-PDT-SURVIVED GLIOBLASTOMA CELLS ARE SUPPRESSED BY USING MEK1/2 INHIBITOR TRAMETINIB
Authors: Kobayashi Tatsuya , Muragaki Yoshihiro , Takahashi Masamichi , Wada Kojiro , Mori Kentaro , Kawamata Takakazu , Ichimura Koichi , Tomiyama Arata 
Published: December 2019

Abstract

INTRODUCTION
In this study, we tried to investigate alteration of oncogenic properties and their molecular regulatory mechanism of talaporfin sodium (NPe6)-mediated photodynamic therapy (NPe6-PDT)-survived glioblastoma (GBM) cells.

METHODS
As the in-vitro NPe6-PDT model, human GBM cell lines (T98G, U87MG, U343), and patient derived GBM stem cells (GSY03, GSC23, MGG152) were pretreated with 0-30ug/ml NPe6 for 4 hours followed by laser irradiation (wave length 664 nm, laser-power 33 mW/cm2, total amount of irradiation 10 J/cm2) using a semiconductor laser irradiator (Panasonic Healthcare Co., Ltd., Tokyo, Japan). Cell death after PDT was evaluated by vital dye exclusion assay using Hoechst3342 and propidium iodide or CellTiter-Glo. Survived cells after NPe6-PDT (PDT-R cells) were repropagated, and alteration of intracellular molecular signaling or migration/invasion capability were analyzed by immunoblotting or Boyden chamber assay.

RESULTS
In both human GBM cell lines and patient derived GBM cells, cellular viability after NPe6-PDT was decreased with dose-dependent manner of pretreated NPe6. PDT-R cells showed not only resistance against NPe6-PDT-induced cell death but also higher invasiveness and migration capability compared with pre-PDT treated cells (PDT-Con cells), and immunoblot analysis demonstrated upregulation of ERK1/2 phosphorylation in PDT-R cells in comparison with PDT-Con cells. Furthermore, these acquired malignant behavior of PDT-R cells were repressed by concomitant use of MEK1/2 inhibitor Trametinib with NPe6-PDT.

CONCLUSION
We discovered PDT-R cells demonstrated higher malignant phenotypes via ERK1/2-dependent machinery compared with parent pre-PDT-treated cells. It was also suggested concomitant treatment with MEK1/2 inhibitor during PDT therapy in GBM cases would contribute to better outcome.
ET-12 ANTI-VEGF THERAPY WITH KETOGENIC DIET AGAINST GLIOBLASTOMA IN MOUSE MODEL
Authors: Sasayama Takashi , Maeyama Masahiro , Tanaka Kazuhiro , Fujita Yuichi , Hashiguchi Mitsuru , Irino Yasuhiro , Kohmura Eiji 
Published: December 2019

Abstract

NTRODUCTUION
Malignant glioma cells critically depend on glucose as the main energy source to survive and sustain their aggressive properties. The ketogenic diet (KD) has been proposed as a complementary therapy for treatment of malignant gliomas. VEGF inhibitor (bevacizumab) decreases blood supply to tumor and clinically used for glioblastoma treatment. Therefore, we examined anti-tumor effect of the combination of bevacizumab (Bev) and KD using mouse model.

METHODS
U87MG cells were implanted into the right brain of nude mice. One week after the implantation, mice were randomized into four treatment groups: control group, KD group, Bev group, and combination (K+B) group. KetoCal 4:1 was administered to the mice for KD. Bev (10mg/kg) was injected from tail vein twice a week. Metabolic and histological analysis of the tumor, and survival analysis of the mice were performed.

RESULTS
3-hydroxy-butyrate, one of the ketone bodies, was significantly increased in the tumor of KD group, however, the metabolic enzymes of ketone bodies were not found an increased expression in immunostaining experiments. Principal component analysis (PCA) analysis demonstrated distinct clustering or a clear separation of the four groups. In K+B group, several TCA cycle-related enzymes (succinate dehydrogenase (SDH), fumarate-hydratase (FH)) were decreased, suggesting a repression of TCA cycle. In addition, several amino acids (tyrosine, valine, alanine, glutamic acid) were decreased in K+B tumor, however, alpha-ketoglutarate was significantly increased, suggesting dynamic metabolic remodeling. Histologically, Ki-67 index was most decreased in the K+B tumor among four groups. In survival analysis, Bev group had significant longer survival than control group (p=0.0016), and the K+B group had most longer survival time among four groups.

CONCLUSIONS
Drastic metabolic remodeling in the tumor occurred in the combination of Bev and KD This combination may be potentially useful for glioblastoma therapy.
IM-03 IMMUNOLOGICAL SUBTYPES OF GLIOBLASTOMA BASED ON TUMOR INFILTRATING CELLS
Authors: Yoshihiro Kushihara , Tanaka Syota , Yamasawa Erika , Koike Tsukasa , Hana Taijun , Kitagawa Yousuke , Takayanagi Syunsaku , Kakimi Kazuhiro , Saito Nobuhito 
Published: December 2019

Abstract
To discover novel biological targets in glioblastoma, genomic and immunological analysis were performed using The Cancer Genome Atlas (TCGA) data set. The RNA-seq data of 156 primary glioblastoma cases were subjected to CIBERSORT to detect tumor infiltrating cell fractions. Principal component analysis was performed on this data to detect factors that strongly contribute to the first principal component, and hierarchical clustering was performed. Survival curves were compared for each of the derived clusters. Finally, Gene Set Enrichment Analysis (GSEA) using HALLMARK Gene Set was performed. In the principal component analysis, we detected seven factors (NK cells resting, T cell regulatory, NK cells activated, Macrophage type 0, T cell gamma delta, Macrophage type 2, Macrophage type 1) which strongly contribute to the first principal component. Based on these seven factors, hierarchical cluster analysis resulted in T cell regulatory (Treg), Macrophage type 0 (M0), Macrophage type 2 (M2) and Macrophage type 1 (M1) clusters. There was no significant difference between these groups in CD8 T cell. M2 and M1 clusters displayed better OS with a significant difference. TNFA signaling via NFκB in Treg group, IFNα response, IFNγ response and ALLOGRAFT response in M2 group, G2M CHECKPOINT, GLYCOLYSIS, WNTβ catenin signaling, MITOTIC SPINDLE and TGFβ signaling in M1 group were upregulated. In conclusion, tumor microenvironment of glioblastoma can be divided into 4 immunological subtypes, Treg, M0, M1, and M2. Because of the contribution of innate immunity for shaping the tumor microenvironment of glioblastoma, immunotherapies targeting these innate immune cells are anticipated.
ACT-01 THE SECOND GENERATION ANAPLASTIC LYMPHOMA KINASE (ALK) INHIBITOR CERITINIB EFFECTIVELY INDUCES CELL DEATH IN HUMAN GLIOBLASTOMA CELLS
Authors: Kawauchi Daisuke , Takahashi Masamichi , Yamamuro Shun , Kobayashi Tatsuya , Uchida Eita , Iwadate Yasuo , Ichimura Koichi , Tomiyama Arata 
Published: December 2019

Abstract
Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase which expresses only in the developmental stage of the brain during embryogenesis of human. On the other hand, a variety of ALK gene alterations, such as oncogenic fusion, activating point mutation, or wild type gene amplification, have been recently discovered as the powerful oncogene in various tumors, and these ALK mutations have also been known as the potential therapeutic targets against tumors harboring these ALK mutations. For example, ALK inhibitors have been already approved and used for the clinical treatment of non-small cell lung cancers harboring oncogenic ALK fusion.
Previously, we reported classical ALK inhibitors triggered cell death in human glioblastoma (GBM) cells, which did not express ALK, via suppression of transcription factor STAT3 activation but not in normal tissue-derived cells.
In this study, we investigated the anti-tumor effect of newly-developed ALK inhibitors in GBM cells. As a result, a second generation ALK inhibitor ceritinib induced cell death in various human GBM cell lines with lower concentration compared to other ALK inhibitors. Besides, ceritinib also suppressed STAT family activity in these GBM cell lines. From these results, we consider ceritinib might be a novel therapeutic agent against GBMs, and further investigation about the specific anti-tumor mechanism of ceritinib in GBM cells is currently on-going.
ACT-09 RETROSPECTIVE INVESTIGATION ABOUT STATUS AND RESULT OF ADMINISTRATION OF BEVACIZUMAB FOR MALIGNANT GLIOMAS IN THE REAL WORLD
Authors: Hashiba Tetsuo , Ueno Katsuya , Naito Nobuaki , Miyata Mayuko , Yamamura Natsumi , Lee Ichi , Iwata Ryoichi , Takeda Junnichi , Yoshimura Kunikazu , Nonaka Masahiro , Asai Akio 
Published: December 2019

Abstract
6 years have passed after approval of Bevacizumab for malignant gliomas in Japan, we analyzed the application and the results in our institution. Subjects were 56 patients who were histologically diagnosed as malignant gliomas. Bevacizumab was used in 41 patients among them. In 14 patients, Bevacizumab was introduced after initial therapy. The resection rates were below partial resection in 11 of the 14 patients. In 12 patients, administrations were finished and the average use was 7.6 times. The reason was PD in 6, and side effect in 4. Eight patients died, the average OS of those who died was 9.9 months, the average PFS after Bevacizumab was 5.4 months, and the average time from discontinuation to death was 2.1 months. In 27 patients used at the time of recurrence, the initial excision rate tended to be higher than in the former cases. In 22 patients the administrations were finished and the average use was 11.1 times. The reason was PD in 17, and side effect in 4. Twenty patients have died, the average OS of those who died was 22.3 months, the average PFS after Bevacizumab was 7.1 months, and the average time from discontinuation to death was 2.6 months. In 12 of 15 unused patients subtotal or total resections were achieved. From results, when it is difficult to control by surgery or TMZ, Bevacizumab is used in most patients, and considering the nature of tumor, it can be said that all patients will be considered for use sometime. However, PFS after introduction is not good and the prognosis after discontinuation is poor. It is necessary to conduct initial treatment that can delay introduction, to provide care that does not lead to discontinuation due to side effects, and to examine what treatment is possible at the time of exacerbation.
STMO-01 APPLICATION OF THE AMINOLEVULINIC ACID HYDROCHLORIDE FOR INTRAOPERATIVE DETECTION OF MALIGNANT BRAIN TUMORS
Authors: Kobayashi Keiichi , Saito Kuniaki , Shimada Daisuke , Matsumoto Yoshie , Shimizu Saki , Iijima Shouhei , Noguchi Akio , Shibahara Jyunji , Shiokawa Yoshiaki , Nagane Motoo 
Published: December 2019

Abstract

BACKGROUND
A maximal safe resection has been shown as an independent prognosis factor for high grade glioma (HGG). Intraoperative photo dynamics diagnosis (PDD) facilitates an increased rate of tumor resection, thereby taking an important role in accomplishment of “ maximal safe resection “ along with neuro-navigation and monitoring of motor nerve function. Since the approval of aminolevulinic acid (5-ALA) for PDD in 2013 in Japan, we have been utilizing the PDD when HGG is suspected by preoperative assessments. Here we retrospectively analyze clinical findings in PDD-mediated tumor resection.

METHODS
From February 2014 to March 2019, 285 consecutive patients (132 females) with suspected HGG underwent total of 302 PDD-mediated resection. Median age was 61 yo (13–90). A single oral dose of 5-ALA 20 mg/kg was given within three hours before microsurgery. Positivity of excited fluorescence was assessed qualitatively under surgical microscope.

RESULTS
Among 195 gliomas, the fluorescence positivity rates were 89.6% (120/134) for glioblastoma, 40.5% (15/37) for grade III, and 13.6% (3/22) for grade II. The positive rates for other histologies were 30.2% (13/43) for malignant lymphoma, 32.1% (9/28) for metastatic brain tumor, 0% (0/7) for meningioma (include atypical), and 4.5% (1/22) for other types. In gliomas the high positivity correlated with histological grades, while all fluorescence-positive grade II gliomas were recurrent tumors. Serious adverse events were not observed.

CONCLUSIONS
The qualitative PDD showed a clinical utility to aid accurate resection in glioblastoma, whereas its positivity was inconsistent in lower grade gliomas as well as other malignant brain tumors. This unreliability might attribute, at least in part, to fluorescent intensity heterogeneity in situ, a reduced excitation by an inappropriate radiation angle of the light source, and the qualitative fluorescence detection. Impacts of introducing quantitative detection system as well as molecular and histopathological features on better discrimination will be discussed.
STMO-05 SURGICAL AND FUNCTIONAL OUTCOME OF AWAKE SURGERY FOR INSULAR GLIOMA
Authors: Hirono Seiichiro , Matsutani Tomoo , Iwadate Yasuo 
Published: December 2019

Abstract

BACKGROUND
Insular glioma is still challenging entity for neurosurgeons because of its deep location and the surrounding vascular structures and eloquent white matter fibers. Objective. To clarify the surgical and neurofunctional outcome of awake surgery for patients with insular glioma (IG).

METHODS
We conducted a retrospective review of 42 consecutive patients with insular glioma (IG) and non-insular glioma (non-IG) who underwent awake craniotomy. In addition to surgical outcome including operative complication, the detailed objective neurocognitive evaluation were also analyzed.

RESULTS
Ten IG and 32 non-IG patients were included in this study. Preoperative tumor volume in IG group was smaller than non-IG group (15.3cc, 21.9cc, p=0.14). More patients (8 in 12) in IG patients had tumor in the right hemisphere than non-IG group (12 out of 32, p=0.06). All the resection were performed according to the functional boundary decided by the cortical and subcortical mapping. Preoperatively, IG patients showed better cognitive function in the TMT-B and categorical fluency test (p=0.035, p=0.026, respectively).Postoperatively, the median residual volume and the resection rate in IG and non-IG group were 2.2cc vs 0cc (p=0.10) and 85.5% vs 100% (p=0.025) respectively, suggesting better resection in non-IG group. In terms of complications, 2 patients (20%) in IG group showed infarction in acute postoperative MR images and mild hemiparesis which recovered completely at 3-month evaluation after operation. In contrast, 16 out of 32 non-IG patients (50%) demonstrated new impaired neurocognitive function, which still persisted at final follow-up in 9 out of 16. Most of their postoperative MR images showed no ischemic lesion, suggesting white matter disconnection as main cause of new deficits. More non-IG patients demonstrated the prolonged time in TMT-A than IG patients at 3-month evaluation.

CONCLUSION
Subcortical white matter disconnection, not ischemic change, are the main reason of postoperative neurocognitive dysfunction in non-IG patients.
MPC-03 IMMUNOHISTOCHEMICAL ANALYSIS OF TUMOR ASSOCIATED MACROPHAGE INDUCED AFTER BIODEGRADABLE CARMUSTINE WAFER IMPLANTATION IN HUMAN GLIOBLASTOMA
Authors: Shibahara Ichiyo , Hagiwara Hiroyuki , Miyasaka Kazuhiro , Sato Sumito , Inukai Madoka , Yasui Yoshie , Hide Takuichiro , Kumabe Toshihiro 
Published: December 2019

Abstract
The carmustine (BCNU) wafer, a biodegradable polymer, currently is the only drug that is able to be placed at the surgical site to treat malignant tumors. Biomaterials to treat cancers hold therapeutic potential; however, how they behave inside the tumor microenvironment requires further study. We previously investigated the tumor microenvironment after BCNU wafer implantation, and found that CD68-positive macrophage was significantly introduced around the wafer (Shibahara et al. J Neurooncol 2018). Recent studies demonstrated the importance of tumor-associated macrophage (TAM). However, we could not clarify whether the increased macrophage around the wafer was pro-tumor or anti-tumor phenotype. In the present study, we immunohistochemically examined expressions of CD68, IBA1, CD163, TMEM119, BIN1, CD31, and VEGF to investigate TAM after the wafer implantation. Quantitative evaluation revealed that CD68-positive cells were significantly increased (P = 0.0009), whereas TMEM119-positive cells were significantly decreased (P = 0.0081) after wafer implantation compared to tissue from cases without wafer implantation. CD163, a known marker of poor prognosis in glioblastoma, did not differ with and without wafer implantation. Among factor analyzed, BCNU wafer did not induce protumor TAM, but reduced microglial marker, TMEM119. In addition to the aspect of chemotherapy, BCNU wafer may have potential to modify the tumor microenvironment such as TAM.
MPC-04 MOLECULAR FEATURES AND CLINICAL OUTCOMES OF ELDERLY GLIOBLASTOMA PATIENTS: ANALYSES OF KANSAI NETWORK AND TCGA COHORTS
Authors: Fukai Junya , Arita Hideyuki , Umehara Toru , Yoshioka Ema , Shofuda Tomoko , Kodama Yoshinori , Kinoshita Manabu , Okita Yoshiko , Nonaka Masahiro , Uda Takehiro , Sakamoto Daisuke , Mori Kanji , Kanemura Yonehiro 
Published: December 2019

Abstract

INTRODUCTION
Aging is a negative prognostic factor in glioblastoma (GB) and the genetic background in clinical outcome of elderly GB could exist. This study investigates the difference of elderly patients from younger ones regarding molecular characteristics as well as clinical outcomes in IDH-wildtype GB.

METHODS
We collected adult cases diagnosed with IDH-wildtype GB and enrolled in Kansai Molecular Diagnosis Network for CNS Tumors (Kansai Network) (212 cases) and The Cancer Genome Atlas (TCGA) project (359 cases). Clinical and pathological characteristics were analyzed retrospectively and compared between elderly cases (≥70 years) and younger ones (≤50 years). Molecular analysis included copy number alterations (CNAs) of eight genes (EGFR, PDGFRA, PTEN, CDKN2A, CDK4, MDM2, TP53, NFKBIA).

RESULTS
Included in the study were 92 (≥70 years)/33 (≤50 years) cases of Kansai Network and 88 (≥70 years)/69 (≤50 years) cases of TCGA. Median overall survival was 12.8 (≥70 years)/ 21.0 (≤50 years) months in Kansai Network cohort and 8.8 (≥70 years)/ 21.09 (≤50 years) months in TCGA cohort. MGMT promoter was methylated in 50 (54.3%) (≥70 years)/14 (42.4%) (≤50 years) tumors in Kansai Network and 34 (48.6%) (≥70 years)/16 (36.4%) (≤50 years) tumors in TCGA. TERT promoter was mutated in 51 (55.4%) (≥70 years)/13 (39.4%) (≤50 years) tumors in Kansai Network and unknown in TCGA. Significant difference of CNA profiles between ≥70 years and ≤50 years was as follows: PTEN del, 43 (46.7%)/8 (24.2%); CDK4 amp, 17 (18.5%)/1 (3.0%) in Kansai Network and CDKN2A del, 69 (78.4%)/ 42 (60.9%) in TCGA.

CONCLUSION
Elderly patients have several potential factors for poor prognosis and different molecular profiles might explain the survival differences among generations.