Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
DAISUKE KUGA Last modified date:2021.05.24

Assistant Professor / Neurosurgery / Kyushu University Hospital

1. Funakoshi Y, Hata N, Kuga D, Hatae R, Sangatsuda Y, Fujioka Y, Takigawa K, Mizoguchi M., Update on Chemotherapeutic Approaches and Management of Bevacizumab Usage for Glioblastoma., Pharmaceuticals , 16;13(12):470, 2020.12.
2. Masaoki Kusunoki, Kazufumi Kikuchi, Osamu Togao, Koji Yamashita, Daichi Momosaka, Yoshitomo Kikuchi, Daisuke Kuga, Nobuhiro Hata, Masahiro Mizoguchi, Koji Iihara, Satoshi O. Suzuki, Toru Iwaki, Yuta Akamine, Akio Hiwatashi, Differentiation of high-grade from low-grade diffuse gliomas using diffusion-weighted imaging
a comparative study of mono-, bi-, and stretched-exponential diffusion models, Neuroradiology, 10.1007/s00234-020-02456-2, 62, 7, 815-823, 2020.07, Purpose: Diffusion-weighted imaging (DWI) plays an important role in the preoperative assessment of gliomas; however, the diagnostic performance of histogram-derived parameters from mono-, bi-, and stretched-exponential DWI models in the grading of gliomas has not been fully investigated. Therefore, we compared these models’ ability to differentiate between high-grade and low-grade gliomas. Methods: This retrospective study included 22 patients with diffuse gliomas (age, 23–74 years; 12 males; 11 high-grade and 11 low-grade gliomas) who underwent preoperative 3 T-magnetic resonance imaging from October 2014 to August 2019. The apparent diffusion coefficient was calculated from the mono-exponential model. Using 13 b-values, the true-diffusion coefficient, pseudo-diffusion coefficient, and perfusion fraction were obtained from the bi-exponential model, and the distributed-diffusion coefficient and heterogeneity index were obtained from the stretched-exponential model. Region-of-interests were drawn on each imaging parameter map for subsequent histogram analyses. Results: The skewness of the apparent diffusion, true-diffusion, and distributed-diffusion coefficients was significantly higher in high-grade than in low-grade gliomas (0.67 ± 0.67 vs. − 0.18 ± 0.63, 0.68 ± 0.74 vs. − 0.08 ± 0.66, 0.63 ± 0.72 vs. − 0.15 ± 0.73; P = 0.0066, 0.0192, and 0.0128, respectively). The 10th percentile of the heterogeneity index was significantly lower (0.77 ± 0.08 vs. 0.88 ± 0.04; P = 0.0004), and the 90th percentile of the perfusion fraction was significantly higher (12.64 ± 3.44 vs. 7.14 ± 1.70%: P < 0.0001), in high-grade than in low-grade gliomas. The combination of the 10th percentile of the true-diffusion coefficient and 90th percentile of the perfusion fraction showed the best area under the receiver operating characteristic curve (0.96). Conclusion: The bi-exponential model exhibited the best diagnostic performance for differentiating high-grade from low-grade gliomas..
3. Daisuke Kuga, Masahiro Toda, Kazunari Yoshida, In Reply to the Letter to the Editor Regarding “Treatment Strategy for Tuberculum Sellae Meningiomas Based on Preoperative Radiologic Assessment”, World Neurosurgery, 10.1016/j.wneu.2019.01.260, 125, 2019.05.
4. Kenji Miki, Koji Yoshimoto, Yuichi Yamada, Akira Kabashima, Daisuke Kuga, Yoshinao Oda, Koji Iihara, Rapid growth of metastatic brain tumor from gastric undifferentiated pleomorphic sarcoma
A case report, Surgical Neurology International, 10.25259/SNI-84-2019, 10, 74, 1-4, 2019.04, Background: Brain metastasis from undifferentiated pleomorphic sarcoma (UPS) is a rare occurrence, and its clinical course is little known. In this report, we investigate a case of a rapidly growing brain metastasis from gastric UPS. Case Description: An 82-year-old man with a known gastric tumor, pathologically compatible with UPS, underwent partial gastrectomy at an outside facility. 3 months later, a 4-cm brain tumor was detected, which was completely resected. The patient was diagnosed with metastatic tumor from previously treated gastric UPS. Within 2 months of the initial resection, a large recurrent mass was detected in the same location, which was again removed. Although the patient underwent radiotherapy and chemotherapy for other metastatic tumors, he died 5 months after the second craniotomy. Conclusions: Brain metastasis from gastric UPS is rare and difficult to treat. Although aggressive treatment, such as surgical intervention, may improve patient survival in some cases, the timing of treatment is challenging because cerebral metastasis rapidly grows and and patients frequently suffer from synchronous systematic metastasis. Therefore, early detection and close follow-up of rapidly progressing brain metastasis are important to improve treatment outcomes..
5. Yuhei Michiwaki, Nobuhiro Hata, Toshiyuki Amano, Satoshi O. Suzuki, Yojiro Akagi, Daisuke Kuga, Daisuke Onozuka, Seiya Momosaki, Akira Nakamizo, Koji Yoshimoto, Toru Iwaki, Koji Iihara, Predictors of recurrence and postoperative outcomes in patients with non-skull base meningiomas based on modern neurosurgical standards, Interdisciplinary Neurosurgery: Advanced Techniques and Case Management, 10.1016/j.inat.2018.10.007, 15, 30-37, 2019.03, Background: Advances in neurosurgical techniques and neuroimaging resolution questions the modern-day reliability of the Simpson grade for predicting meningioma recurrence. Therefore, we evaluated the reliability of predictors for recurrence and outcomes in detail in patients with non-skull base meningiomas (NSBMs). Methods: We retrospectively analyzed data from consecutive 175 NSBMs underwent surgical resection. We performed Kaplan–Meier analyses of recurrence-free survival (RFS) according to Simpson and World Health Organization (WHO) grades. Predictors of RFS and clinical deterioration were estimated by univariate and multivariate analyses. Correlation between the Simpson grade and change in Karnofsky Performance Scale scores was assessed by Fisher's exact test. Results: Log-rank tests revealed significant correlations of both the Simpson and WHO grades with RFS for the overall cohort, convexity, and falx/tentorium meningioma. Unlike patients undergoing Simpson grade I and II resections, RFS in patients with WHO grade I and II/III tumors differed significantly from the early postoperative stage. Multivariate analysis identified tumor size, Simpson grade, and MIB-1 labeling index as significant predictors of RFS. Clinical deterioration was more frequent among patients undergoing less aggressive resection. Tumor location was the only significant predictor of clinical deterioration. Conclusions: Our findings indicate that tumor size, Simpson and WHO grades, and MIB-1 labeling index are significant predictors of NSBM recurrence. Moreover, the risk of recurrence markedly decreases within the follow-up duration of 80 months. Aggressive resection appears to minimize the risk of recurrence without evidence of clinical deterioration. Follow-up schedules should be based on the WHO grade and extent of resection..
6. Daisuke Kuga, Masahiro Toda, Hiroyuki Ozawa, Kaoru Ogawa, Kazunari Yoshida, Endoscopic Endonasal Approach Combined with a Simultaneous Transcranial Approach for Giant Pituitary Tumors, World Neurosurgery, 10.1016/j.wneu.2018.10.047, 121, 173-179, 2019.01, Background: The endoscopic endonasal approach is widely used for treating giant pituitary adenomas. However, a small subset of tumors is still challenging to treat, and the risk of complications increases when an endoscopic endonasal approach alone is used. The simultaneous combined endoscopic endonasal and transcranial approach is a surgical option for such difficult adenomas; however, very few studies have described the technical nuances and benefits of this approach. Methods: We treated 3 patients with giant pituitary adenoma and 1 patient with pituicytoma. Radiologic findings and clinical outcomes were retrospectively reviewed. Results: All patients had preoperative visual disturbances. A pterional approach was combined with an endoscopic endonasal approach to treat all the patients. Near-total and subtotal tumor removal was accomplished in 3 patients; however, only partial tumor removal was possible in 1 patient. Postoperative visual function improved in 3 patients, but there were no changes in 1 patient. There were no major complications; however, each patient developed either adrenocorticotropic hormone (ACTH) and thyroid-stimulating hormone deficiency or ACTH deficiency and persistent diabetes. Importantly, no cerebrospinal fluid leakage was observed in the patients. Conclusions: Our simultaneous combined endoscopic and transcranial approach offers safe tumor resection and a low rate of complications. In this procedure, it is important that tumor debulking be performed by the main surgeon via a single surgical route and not by 2 surgeons using the simultaneous endonasal and transcranial approach, to avoid interference in the surgical field. This approach may be considered as a surgical option for carefully selected tumors in the sellar region..
7. K. Yamashita, R. Hatae, Hiwatashi Akio, Osamu Togao, kazufumi kikuchi, D. Momosaka, Y. Yamashita, Daisuke Kuga, Nobuhiro Hata, K. Yoshimoto, S. O. Suzuki, Toru Iwaki, Koji Iihara, Hiroshi Honda, Predicting TERT promoter mutation using MR images in patients with wild-type IDH1 glioblastoma, Diagnostic and Interventional Imaging, 10.1016/j.diii.2019.02.010, 2019.01, Purpose: The purpose of this study was to identify magnetic resonance imaging (MRI) features that are associated with telomerase reverse transcriptase promoter mutation (TERTm) in glioblastoma. Materials and methods: A total of 112 patients with glioblastoma who had MRI at 1.5- or 3.0-T were retrospectively included. There were 43 patients with glioblastoma with wild-type TERT (TERTw) (22 men, 21 women; mean age, 47 ± 25 [SD] years; age range: 3–84 years) and 69 patients with glioblastoma with TERTm (34 men, 35 women; mean age 64 ± 11 [SD] years; age range, 41-–85 years). The feature vectors consist of 11 input units for two clinical parameters (age and gender) and nine MRI characteristics (tumor location, subventricular extension, cortical extension, multiplicity, enhancing volume, necrosis volume, the percentage of necrosis volume, minimum apparent diffusion coefficient [ADC] and normalized ADC). First, the diagnostic performance using univariate and multivariate logistic regression analyses was evaluated. Second, the cross-validation of the support vector machine (SVM) was performed by using leave-one-out method with 43 TERTw and 69 TERTm to evaluate the diagnostic performance. In addition, the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy for the differentiation between TERTw and TERTm were compared between logistic regression analysis and SVM. Results: With multivariate analysis, the percentage of necrosis volume and age were significantly greater in TERTm glioblastoma than in TERTw glioblastoma. SVM allowed discriminating between TERTw glioblastoma and TERTm glioblastoma with sensitivity, specificity, PPV, NPV, and accuracy of 85.7% [60/70; 95% confidence interval (CI): 75.3–92.9%], 54.8% (23/42; 95% CI: 38.7–70.2%), 75.9% (60/79; 95% CI: 69.1–81.7%), 69.7% (23/33; 95% CI: 54.9–81.3%) and 74.1% (83/112; 95% CI: 65.0–81.9%), respectively. Conclusion: The percentage of necrosis volume and age may surrogate for predicting TERT mutation status in glioblastoma..
8. Daisuke Kuga, Masahiro Toda, Kazunari Yoshida, Treatment Strategy for Tuberculum Sellae Meningiomas Based on a Preoperative Radiological Assessment, World Neurosurgery, 10.1016/j.wneu.2018.09.054, 120, e1279-e1288, 2018.12, Background: Although there are several surgical approaches for the treatment of tuberculum sellae (TS) meningiomas, clear indications for non–large TS meningiomas are still lacking. Methods: Our case series included 20 patients with TS meningiomas (<3 cm). We classified the tumors into 3 groups based on their radiologic relationship with the optic chiasm: type I, tumor with intact optic chiasm; type II, tumor with superiorly deviated optic chiasm; and type III, tumor with posteriorly deviated optic chiasm. Clinical outcomes, radiologic findings, and surgical approaches for the removal of each tumor type were retrospectively reviewed. Results: Resections using a pterional approach, interhemispheric approach, and an endoscopic endonasal approach were performed in three groups of 6, 7, and 7 patients. The rate of total tumor resection was equivalent across approaches, whereas postoperative visual dysfunction was observed in 1 patient (7.69%) undergoing a transcranial approach. Our evaluation of the sphenoid sinus shape across radiographs revealed that the patterns of bony wall elongation attached to these tumors significantly differed among tumor types, indicating that tumor origin and growth direction might affect the patterns of optic chiasm deviation. In addition, selective elongation of the TS provided a favorable surgical corridor for an endoscopic endonasal approach, especially in type II tumors. These results indicate that this tumor classification influenced surgical approach selection for non–large TS meningiomas. Conclusions: The aim of surgery is maximal tumor resection without causing visual dysfunction. The classification proposed here may predict surgical risk associated with meningioma resection and further inform the selection of a surgical approach..
9. yuhei sangatsuda, Nobuhiro Hata, Satoshi Suzuki, Yojiro Akagi, Ryusuke Hatae, Daisuke Kuga, Koji Yoshimoto, Seiya Momosaki, Toru Iwaki, Koji Iihara, An elderly case of malignant small cell glioma with hemorrhage coexistent with a calcified pilocytic astrocytoma component in the cerebellar hemisphere, Neuropathology, 10.1111/neup.12478, 38, 5, 493-497, 2018.10, Pilocytic astrocytoma is a less aggressive form of glial tumor that commonly occurs in the pediatric population, and its malignant transformation is extremely rare. Here, we report an elderly case of malignant small cell glioma with hemorrhage coexistent with a calcified pilocytic astrocytoma component. An 80-year-old male was found to have a right cerebellar non-enhanced tumor with hematoma adjoining a calcified nodule. The lesion was surgically removed, and a histological examination verified that the tumor was a malignant small cell glioma with hemorrhagic change and the calcified nodule showed features of pilocytic astrocytoma. Genetic analyses revealed no glioma-relevant genetic alterations such as IDH and BRAF mutations. Although calcification is generally observed in slowly growing gliomas, the aggressive clinical course of calcified cerebellar pilocytic astrocytoma has been previously reported. Our extremely rare case shows that careful follow-up is necessary even for calcified pilocytic astrocytomas..
10. Tomohiro Okuda, Nobuhiro Hata, Satoshi Suzuki, Koji Yoshimoto, Koichi Arimura, Takeo Amemiya, Yojiro Akagi, Daisuke Kuga, Utako Oba, Yuhki Koga, Shoichi Ohga, Toru Iwaki, Koji Iihara, Pediatric ganglioglioma with an H3 K27M mutation arising from the cervical spinal cord, Neuropathology, 10.1111/neup.12471, 38, 4, 422-427, 2018.08, The 2016 edition of the World Health Organization Classification of Tumors of the Central Nervous System introduced “diffuse midline glioma H3 K27M mutant” as a new diagnostic entity. These tumors predominately affect pediatric patients and arise from midline structures such as the brainstem, thalamus and spinal cord. Here, we report a rare patient with spinal ganglioglioma carrying an H3 K27M mutation. A 10-year-old boy presented with an intramedullary tumor in the cervical spinal cord. The lesion was partially removed and histologically diagnosed as ganglioglioma. After the remnant tumor grew within 3 months after surgery, the patient underwent radiotherapy. Genetic analyses revealed an H3F3A K27M mutation but no other genetic alterations such as IDH and BRAF mutations. This case may point to pathological heterogeneity in gliomas with H3 K27M mutations..
11. Daisuke Kuga, Nobuhiro Hata, Yojiro Akagi, Takeo Amemiya, yuhei sangatsuda, Ryusuke Hatae, Koji Yoshimoto, Masahiro Mizoguchi, Koji Iihara, The Effectiveness of Salvage Treatments for Recurrent Lesions of Oligodendrogliomas Previously Treated with Upfront Chemotherapy, World Neurosurgery, 10.1016/j.wneu.2018.03.069, 114, e735-e742, 2018.06, Background: We previously reported a favorable outcome in a case series of patients with oligodendrogliomas treated with upfront chemotherapy; however, their progression-free survival (PFS) was relatively short considering their long-term overall survival (OS). This suggests that salvage treatments after progression were effective. However, the clinical impact of salvage treatments on outcomes of patients with recurrent oligodendrogliomas has not been precisely investigated. Methods: Our case series included 28 patients with newly diagnosed isocitrate dehydrogenase–mutant and 1p/19q-codeleted oligodendroglial tumors treated with upfront procarbazine, nimustine, and vincristine. Clinical outcomes and patterns of recurrence were reviewed retrospectively. Results: The median follow-up period of enrolled patients was 90.2 months. Disease progression occurred in 15 patients (53.6%), whereas the cancer appeared as local relapse alone in 14 (93.3%) patients. Salvage treatments were performed for all local relapses; thereafter, most of the subsequent progressions also appeared as resectable local relapses. The 5-year PFS and OS rates from the first progression were 30.3% and 92.9%, respectively. These relatively short PFS and favorable OS indicated the effectiveness of salvage treatment even after multiple progression. Thus far, 9 (60%) of 15 patients are deterioration-free with locally controlled lesions or complete remission; however, clinical deterioration was observed in 6 patients, and 4 of them experienced dissemination. Conclusions: In isocitrate dehydrogenase–mutant and 1p/19q-codeleted oligodendrogliomas, most of the tumors that demonstrated early progression appeared as local, nonlethal lesions, which have been well-controlled by salvage treatments. A precise diagnosis of oligodendrogliomas using molecular parameters is crucial to receive the best benefit from salvage treatment..
12. Yojiro Akagi, Koji Yoshimoto, Nobuhiro Hata, Daisuke Kuga, Ryusuke Hatae, Takeo Amemiya, yuhei sangatsuda, Satoshi Suzuki, Toru Iwaki, Masahiro Mizoguchi, Koji Iihara, Reclassification of 400 consecutive glioma cases based on the revised 2016WHO classification, Brain tumor pathology, 10.1007/s10014-018-0313-4, 1-9, 2018.03, In this study, we reclassified 400 consecutive glioma cases including pediatric cases, using the revised 2016 WHO classification with samples collected from the Kyushu University Brain Tumor Bank. The IDH1/2, H3F3A, key genetic markers in the 2016 classification, were analyzed using high-resolution melting, with DNA extracted from frozen tissues. The 1p/19q codeletions were evaluated using a microsatellite-based loss of heterozygosity analysis, with 18 markers, to detect loss of the entire chromosome arm. In the integrated diagnosis, 29 oligodendroglioma cases and 28 anaplastic oligodendroglioma cases were diagnosed as “IDH-mutant and 1p/19q-codeleted,” while 2 oligodendroglioma cases and 5 anaplastic oligodendroglioma cases were diagnosed as not otherwise specified (NOS). These “NOS” cases were either IDH-mutants or 1p/19q-codeleted, although characteristic oligodendroglial features were evident histologically. Better overall survival of patients with oligodendroglioma correlated with the molecular characteristic of “IDH-mutant and 1p/19q-codeleted,” rather than the WHO grade. Eleven “glioblastoma, IDH-wild-type” cases were classified as “1p/19q-codeleted”, however, chromosome 10 loss was also detected in 10 out of 11 cases. The 2016 WHO criteria for glioma classification leads to better diagnosis of patients. However, there are technical pitfalls and problems to be solved in the molecular analysis of routine diagnostics..
13. Koji Yoshimoto, Ryusuke Hatae, Satoshi Suzuki, Nobuhiro Hata, Daisuke Kuga, Yojiro Akagi, Takeo Amemiya, yuhei sangatsuda, Nobutaka Mukae, Masahiro Mizoguchi, Toru Iwaki, Koji Iihara, High-resolution melting and immunohistochemical analysis efficiently detects mutually exclusive genetic alterations of adamantinomatous and papillary craniopharyngiomas, Neuropathology, 10.1111/neup.12408, 38, 1, 3-10, 2018.02, Craniopharyngioma consists of adamantinomatous and papillary subtypes. Recent genetic analysis has demonstrated that the two subtypes are different, not only in clinicopathological features, but also in molecular oncogenesis. Papillary craniopharyngioma (pCP) is characterized by a BRAF mutation, the V600E (Val 600 Glu) mutation. Adamantinomatous craniopharyngioma (aCP) can be distinguished by frequent β-catenin gene (CTNNB1) mutations. Although these genetic alterations can be a diagnostic molecular marker, the precise frequency of these mutations in clinical specimens remains unknown. In this study, we first evaluated BRAF V600E and CTNNB1 mutations in four and 14 cases of pCP and aCP, respectively, using high-resolution melting analysis followed by Sanger sequencing. The results showed that 100% (4/4) of pCP cases had BRAF V600E mutations, while 78% (11/14) of the aCP cases had CTNNB1 mutations, with these genetic alterations being subtype-specific and mutually exclusive. Second, we evaluated BRAF V600E and CTNNB1 mutations by immunohistochemical analysis (IHC). All pCP cases showed positive cytoplasmic staining with the BRAF V600E-mutant antibody (VE-1), whereas 86% (12/14) of aCP cases showed positive cytoplasmic and nuclear staining for CTNNB1, suggesting a CTNNB1 mutation. Only one case of wild-type CTNNB1 on the DNA analysis showed immunopositivity on IHC. We did not detect a coexistence of BRAF V600E and CTNNB1 mutations in any single tumor, which indicated that these genetic alterations were mutually exclusive. We also report our modified IHC protocol for VE-1 staining, and present the possibility that BRAF V600E mutations can be used as a diagnostic marker of pCP in the differentiation of Rathke cleft cyst with squamous metaplasia..
14. Osamu Togao, Hiwatashi Akio, Koji Yamashita, kazufumi kikuchi, Daichi Momosaka, Koji Yoshimoto, Daisuke Kuga, Masahiro Mizoguchi, Satoshi Suzuki, Toru Iwaki, Marc Van Cauteren, Koji Iihara, Hiroshi Honda, Measurement of the perfusion fraction in brain tumors with intravoxel incoherent motion MR imaging
Validation with histopathological vascular density in meningiomas, British Journal of Radiology, 10.1259/bjr.20170912, 91, 1085, 2018.01, Objective: To evaluate the quantification performance of the perfusion fraction (f) measured with intravoxel incoherent motion (IVIM) MR imaging in a comparison with the histological vascular density in meningiomas. Methods: 29 consecutive patients with meningioma (59.0 ± 16.8 years old, 8 males and 21 females) who underwent a subsequent surgical resection were examined with both IVIM imaging and a histopathological analysis. IVIM imaging was conducted using a singleshot SE-EPI sequence with 13 b-factors (0, 10, 20, 30, 50, 80, 100, 200, 300, 400, 600, 800, 1000 s mm-2) at 3T. The perfusion fraction (f) was calculated by fitting the IVIM bi-exponential model. The 90-percentile f-value in the tumor region-of-interest (ROI) was defined as the maximum f-value (f-max). Histopathological vascular density (%Vessel) was measured on CD31-immunostainted histopathological specimens. The correlation and agreement between the f-values and %Vessel was assessed. Results: The f-max (15.5 ± 5.5%) showed excellent agreement [intraclass correlation coefficient (ICC) = 0.754] and a significant correlation (r = 0.69, p < 0.0001) with the %Vessel (12.9 ± 9.4%) of the tumors. The Bland- Altman plot analysis showed excellent agreement between the f-max and %Vessel (bias, -2.6%; 95% limits of agreement, from -16.0 to 10.8%). The f-max was not significantly different among the histological subtypes of meningioma. Conclusion: An excellent agreement and a significant correlation were observed between the f-values and %Vessel. The f-value can be used as a noninvasive quantitative imaging measure to directly assess the vascular volume fraction in brain tumors. Advances in knowledge: The f-value measured by IVIM imaging showed a significant correlation and an excellent agreement with the histological vascular density in the meningiomas. The f-value can be used as a noninvasive and quantitative imaging measure to directly assess the volume fraction of capillaries in brain tumors..
15. Koji Yoshimoto, Ryusuke Hatae, Yuhei Sangatsuda, Satoshi Suzuki, Nobuhiro Hata, Yojiro Akagi, Daisuke Kuga, Murata Hideki, Koji Yamashita, Osamu Togao, Hiwatashi Akio, Toru Iwaki, Masahiro Mizoguchi, Koji Iihara, Prevalence and clinicopathological features of H3.3 G34-mutant high-grade gliomas
a retrospective study of 411 consecutive glioma cases in a single institution, Brain Tumor Pathology, 10.1007/s10014-017-0287-7, 34, 3, 103-112, 2017.07, A recurrent glycine-to-arginine/valine alteration at codon 34 (G34R/V) within H3F3A, a gene that encodes the replication-independent histone variant H3.3, reportedly occurs exclusively in pediatric glioblastomas. However, the clinicopathological and biological significances of this mutation have not been completely elucidated; especially, no such data exist for tumor samples from Japanese patients. We analyzed 411 consecutive glioma cases representing patients of all ages. Our results demonstrated that 14 patients (3.4%) harbored H3F3A mutations, of which four had G34R mutations and 10 had K27M mutations. G34R-mutant tumors were located in the parietal region in two patients and the basal ganglia in one patient. One patient showed multi-lobular extension similar to the pattern observed in gliomatosis cerebri. Regarding neuroradiological features, intratumoral calcification was evident in two cases and all cases showed no or scarce contrast enhancement on MRI. Histopathologically, the four G34R-mutant cases included three glioblastomas and one astroblastoma. We have also investigated alterations in histone methylation including H3K27me3, H3K9me3, and H3K4me3 in G34R-mutant samples by immunohistochemistry. These results indicate that G34R-mutant tumors are likely to show extensive infiltration and alterations in global histone trimethylation might also play an important role in G34R mutant tumors..
16. Ryusuke Hatae, Nobuhiro Hata, Satoshi Suzuki, Koji Yoshimoto, Daisuke Kuga, Hideki Murata, Yojiro Akagi, Yuhei Sangatsuda, Toru Iwaki, Masahiro Mizoguchi, Koji Iihara, A comprehensive analysis identifies BRAF hotspot mutations associated with gliomas with peculiar epithelial morphology, Neuropathology, 10.1111/neup.12347, 37, 3, 191-199, 2017.06, Brain tumors harbor various BRAF alterations, the vast majority of which are the BRAF kinase-activating V600E mutation. BRAF mutations are most frequently detected in certain subtypes of low-grade glioma, such as pilocytic astrocytoma (PA), pleomorphic xanthoastrocytoma (PXA), ganglioglioma (GG) and dysembryoplastic neuroepithelial tumor (DNT). However, it is unclear whether gliomas harboring BRAF mutations can be invariably regarded as these glioma subtypes or their derivatives. To address this question, we analyzed 274 gliomas in our institutional case series. We performed high-resolution melting analyses and subsequent direct Sanger sequencing on DNA isolated from snap-frozen tumor tissues. As expected, BRAF mutations were detected in the aforementioned low-grade gliomas: in 4/27 PAs, 2/3 PXAs, 4/8 GGs, and 1/6 DNTs. In addition to these gliomas, 1/2 astroblastomas (ABs) and 2/122 glioblastomas (GBs) harbored BRAF mutations. Pathological investigation of the two GBs revealed that one was a GB displaying epithelial features that presumably arose from a precedent GG, whereas the other GB, which harbored a rare G596 A mutation, showed marked epithelial features, including astroblastic rosettes. Our results indicate that in addition to being present in established BRAF-associated gliomas, BRAF mutations might be associated with epithelial features in high-grade gliomas, including sheet-like arrangement of polygonal tumor cells with a plump cytoplasm and astroblastic rosettes, and thus could potentially serve as a genetic marker for these features..
17. Nobuhiro Hata, Ryusuke Hatae, Koji Yoshimoto, Hideki Murata, Daisuke Kuga, Yojiro Akagi, Yuhei Sangatsuda, Satoshi Suzuki, Toru Iwaki, Masahiro Mizoguchi, Koji Iihara, Insular primary glioblastomas with IDH mutations
Clinical and biological specificities, Neuropathology, 10.1111/neup.12362, 37, 3, 200-206, 2017.06, Isocitrate dehydrogenase (IDH) mutation is a good prognostic marker for glioblastoma (GBM). Although it is infrequent in primary tumors, it is found in most lower-grade gliomas. Thus, it is unclear whether IDH mutation is a marker for a specific phenotype of apparently primary de novo GBMs (pGBMs), or a marker for secondary tumors (sGBMs). We addressed this issue by analyzing clinical, radiographic and molecular findings in our institutional case series. Our cases included 92 pGBMs, with five cases of IDH1 mutations at R132 and no IDH2 mutations. The median overall survival of these five patients was 29 months (range: 4 to >40 months), which is considered good prognoses. Clinical and radiographic characteristics were distinct from IDH-wildtype (IDH-wt) pGBMs. IDH-mutant (IDH-mut) tumors consistently involved insular lesions and were subdivided into: (i) the two cases of elderly patients with long clinical histories and features implying multistep tumor development; and (ii) the three cases of younger patients with diffusely swelling insular tumors, slight contrast enhancement and no necrosis. Genetic and expression analyses of IDH-mut pGBMs were similar to those of sGBMs, suggesting that they are indeed distinct from their IDH-wt counterparts. TERT promoter mutation, a genetic marker of oligodendroglial derivation, was detected in one long-surviving case, but genetic alterations in the astrocyte-sGBM pathway were generally prevalent in IDH-mut pGBMs. Our results present a unique phenotype of IDH-mut pGBMs arising from insular cortex region, the molecular backgrounds of which are similar to sGBMs..
18. Osamu Togao, Hiwatashi Akio, Koji Yamashita, kazufumi kikuchi, Jochen Keupp, Koji Yoshimoto, Daisuke Kuga, Masami Yoneyama, Satoshi Suzuki, Toru Iwaki, Masaya Takahashi, Koji Iihara, Hiroshi Honda, Grading diffuse gliomas without intense contrast enhancement by amide proton transfer MR imaging
comparisons with diffusion- and perfusion-weighted imaging, European Radiology, 10.1007/s00330-016-4328-0, 27, 2, 578-588, 2017.02, Objectives: To investigate whether amide proton transfer (APT) MR imaging can differentiate high-grade gliomas (HGGs) from low-grade gliomas (LGGs) among gliomas without intense contrast enhancement (CE). Methods: This retrospective study evaluated 34 patients (22 males, 12 females; age 36.0 ± 11.3 years) including 20 with LGGs and 14 with HGGs, all scanned on a 3T MR scanner. Only tumours without intense CE were included. Two neuroradiologists independently performed histogram analyses to measure the 90th-percentile (APT90) and mean (APTmean) of the tumours’ APT signals. The apparent diffusion coefficient (ADC) and relative cerebral blood volume (rCBV) were also measured. The parameters were compared between the groups with Student’s t-test. Diagnostic performance was evaluated with receiver operating characteristic (ROC) analysis. Results: The APT90 (2.80 ± 0.59 % in LGGs, 3.72 ± 0.89 in HGGs, P = 0.001) and APTmean (1.87 ± 0.49 % in LGGs, 2.70 ± 0.58 in HGGs, P = 0.0001) were significantly larger in the HGGs compared to the LGGs. The ADC and rCBV values were not significantly different between the groups. Both the APT90 and APTmean showed medium diagnostic performance in this discrimination. Conclusions: APT imaging is useful in discriminating HGGs from LGGs among diffuse gliomas without intense CE. Key Points: • Amide proton transfer (APT) imaging helps in grading non-enhancing gliomas • High-grade gliomas showed higher APT signal than low-grade gliomas • APT imaging showed better diagnostic performance than diffusion- and perfusion-weighted imaging.
19. Nobuhiro Hata, Koji Yoshimoto, Ryusuke Hatae, Daisuke Kuga, Yojiro Akagi, Yuhei Sangatsuda, Satoshi Suzuki, Tadahisa Shono, Masahiro Mizoguchi, Koji Iihara, Add-on bevacizumab can prevent early clinical deterioration and prolong survival in newly diagnosed partially resected glioblastoma patients with a poor performance status, OncoTargets and Therapy, 10.2147/OTT.S125587, 10, 429-437, 2017.01, Purpose: The AVAglio trial established the beneficial effect of add-on bevacizumab (BEV) for the treatment of newly diagnosed glioblastomas (nd-GBMs) that led to the approval of BEV for the treatment of these patients in Japan. However, the rationality of using BEV as a first-line treatment for nd-GBMs remains controversial. The purpose of this study was to analyze the outcomes of a case series of nd-GBM patients. Patients and methods: The outcomes of 69 nd-GBM patients treated after 2006 were retrospectively analyzed. Clinical and genetic analyses were performed, and estimates of progression-free survival (PFS) and overall survival (OS) were calculated using the Kaplan–Meier method. Since add-on BEV therapy was only used for partially resected GBMs (pr-GBMs) after its approval in 2013, the patients were subdivided into 3 treatment groups: Type I, partial removal with temozolomide (TMZ)/BEV and concurrent radiotherapy (CCRT); Type II, partial removal with TMZ and CCRT; and Type III, gross total removal with TMZ and CCRT. Results: The PFS rate of Type I patients was significantly higher than that of Type II patients (P=0.014), but comparable to that of Type III patients. Differences in OS rates between Type I and Type II patients were less apparent (P=0.075), although the median OS of Type I patients was ∼8 months higher than that of Type II patients (17.4 vs 9.8 months, respectively). The clinical deterioration rate during initial treatment was significantly (P=0.024) lower in Type I than in Type II patients (7.7% vs 47.4%, respectively). Differences in OS rates between Type I and Type II patients with a poor performance status (PS) were significant (P=0.017). Conclusion: Our findings suggest that add-on BEV can prevent early clinical deterioration of pr-GBM patients and contribute to a prolonged survival, especially for those with a poor PS..
20. Satoshi Karashima, Koichi Arimura, Kimiaki Hashiguchi, Yojiro Akagi, Nobutaka Mukae, ataru nishimura, Daisuke Kuga, Koji Yoshimoto, Tetsuro Sayama, Satoshi Suzuki, Koji Iihara, A case of concurrent schwannoma and meningioma at the same cervical level, Japanese Journal of Neurosurgery, 10.7887/jcns.26.750, 26, 10, 750-756, 2017, Concurrent spinal tumors such as schwannomas and meningiomas are usually associated with neurofibromatosis type 2(NF2). Here we report an extremely rare case of concurrent schwannoma and meningioma at the same cervical spinal level. This patient did not meet the diagnostic criteria for NF2. Because the surgical strategy for spinal schwannomas may differ if meningiomas are concurrently present, it is important to pay attention to preoperative imaging findings. Unlike in previous patients, however, it was difficult to diagnose the presence of two different tumors based on preoperative images of the present patient. Preoperative images and intraoperative findings must therefore be carefully assessed to determine whether spinal tumors extending into the intradural and extradural spaces consist of concurrent schwannomas and meningiomas..
21. Koji Yoshimoto, Nobutaka Mukae, Daisuke Kuga, Koji Iihara, Indications and limitations of endoscopic endonasal surgery, Japanese Journal of Neurosurgery, 10.7887/jcns.26.404, 26, 6, 404-411, 2017, The introduction of endoscopy, in conjunction with the technical development of endoscopy such as high definition and 3D, has had a significant impact on the field of endonasal transsphenoidal surgery. Cur­rently, endoscopic surgery is applied for the resection of not only sellar tumors but also parasellar tumors such as anterior skull base tumors, intraorbital tumors, cavernous sinus tumors, infratemporal fossa tumors, clival tumors, and others. Endoscopic endonasal surgery is expected to be used more frequently in the near future. In this paper, based on our own experiences and after reviewing the recent publications, we summarize the indications for endoscopic surgery, focusing on pituitary tumors, craniopharyngiomas, and tuberculum sellae meningiomas. In pituitary tumor surgery, operative results by endoscopic and microscopic surgery are comparable for small tumors located in the sellar, while endoscopic surgery is superior to microscopic surgery for large tumors with extrasellar extension. Fully endoscopic surgery is more likely to be performed for giant pitu­itary adenoma cases for which craniotomy was previously preferred. However, tumors with significant anterior and lateral extension, multilobular suprasellar extension, and firm tumors can be limitations for endoscopic surgery. For craniopharyngiomas, retrochiasmatic tumors extending into the third ventricle can be good candidates for endoscopic surgery. In addition, the tumor-third ventricular relationship in cranio­pharyngiomas is another important factor to consider when deciding upon an indication for endoscopic surgery. The indication for an endoscopic approach for tuberculum sellae meningioma is limited to small anterior midline tumors for the purpose of decompression of the optic apparatus. In summary, although the efficacy of endoscopic surgery is well established, significant suprasellar and lateral extension and vascular encasement can be limitations for endoscopic surgery. For safe and effective endoscopic surgery, patient selection is important depending on the surgeons' operative techniques and experiences. Also, it is important to note that observation and manipulation are different, and it is necessary to acquire surgical techniques to prevent postoperative cerebrospinal fluid leaks..
22. Koji Yoshimoto, Nobutaka Mukae, Daisuke Kuga, Daisuke Inoue, Makoto Hashizume, Koji Iihara, Dual optical channel three-dimensional neuroendoscopy
Clinical application as an assistive technique in endoscopic endonasal surgery, Interdisciplinary Neurosurgery: Advanced Techniques and Case Management, 10.1016/j.inat.2016.08.001, 6, 45-50, 2016.12, Three-dimensional (3D) high-definition endoscopy is an innovative technical advancement that helps surgeons gain precise depth perception and spatial recognition during endoscopic surgery. Here, we describe a new dual optical channel 3D neuroendoscopic technique and its clinical application. We performed endoscopic endonasal surgery on 88 patients using 3D and two-dimensional (2D) endoscopes in conjunction. We evaluated the usefulness of stereoscopic images acquired by dual optical channel 3D endoscopy during endoscopic surgery and compared the image resolution between dual optical channel 3D endoscopy and 2D endoscopy. Additionally, we compared the stereoscopic images acquired by dual optical channel and Visionsense 3D endoscopy in three cases. Combination surgery using 3D and 2D endoscopy was found to be safe. Stereoscopic images were useful in several surgical steps, especially in recognition of complex bony structures, bone drilling, and suprasellar manipulation. The magnitude of binocular disparity was greater in dual optical channel 3D endoscopy than in Visionsense 3D endoscopy. Stereoscopic images acquired by dual optical channel 3D neuroendoscopy were of adequate quality and were useful for endoscopic endonasal surgery. In consideration of its lower image resolution compared to that of 2D high-definition endoscopy, dual optical channel 3D neuroendoscopy can be applied as an assistive technique in endoscopic endonasal surgery. The magnitude of binocular disparity is one of the key factors to be considered for evaluation of the clinical significance of 3D endoscopy..
23. Nobuhiro Hata, Koji Yoshimoto, Ryusuke Hatae, Daisuke Kuga, Yojiro Akagi, Satoshi Suzuki, Toru Iwaki, Tadahisa Shono, Masahiro Mizoguchi, Koji Iihara, Deferred radiotherapy and upfront procarbazine-ACNU-vincristine administration for 1p19q codeleted oligodendroglial tumors are associated with favorable outcome without compromising patient performance, regardless of WHO grade, OncoTargets and Therapy, 10.2147/OTT.S115911, 9, 7123-7131, 2016.11, Recently updated phase III trials revealed the favorable effect of add-on procarbazine-lomustine-vincristine chemotherapy (CT) to radiotherapy (RT) in treating anaplastic oligodendrogliomas with 1p19q codeletion (codel). However, the underlying rationality of deferring RT and upfront CT administration for these tumors is yet to be elucidated. Here, we retrospectively analyzed the long-term outcome of our case series with oligodendroglial tumors treated with deferred RT and upfront procarbazine+nimustine+vincristine (PAV) in the introduction administration. We enrolled 36 patients with newly diagnosed oligodendroglial tumors (17, grade II and 19, grade III) treated during 1999-2012 and followed up for a median period of 69.0 months. Their clinical and genetic prognostic factors were analyzed, and progression-free survival, overall survival (OS), and deterioration-free survival (DFS) were evaluated. Regardless of the WHO grade, the 25 patients with 1p19q codel tumors never received RT initially, and of these 25, 23 received PAV treatment upfront. The 75% OS of patients with 1p19q codel tumor was 135.3 months (did not reach the median OS), indicating a favorable outcome. Multivariate analysis revealed that IDH mutation and 1p19q, not WHO grade, are independent prognostic factors; furthermore, IDH and 1p19q status stratified the cohort into 3 groups with significantly different OS. The DFS explained the prolonged survival without declining performance in patients with both grade II and III 1p19q codel tumors. Deferred RT and upfront PAV treatment for 1p19q codel oligodendrogliomas were associated with favorable outcomes without compromising performance status, regardless of WHO grade..
24. Koji Yamashita, Hiwatashi Akio, Osamu Togao, kazufumi kikuchi, Yoshiyuki Kitamura, Masahiro Mizoguchi, Koji Yoshimoto, Daisuke Kuga, Satoshi Suzuki, Shingo Baba, Takuro Isoda, Toru Iwaki, Koji Iihara, Hiroshi Honda, Diagnostic utility of intravoxel incoherent motion mr imaging in differentiating primary central nervous system lymphoma from glioblastoma multiforme, Journal of Magnetic Resonance Imaging, 10.1002/jmri.25261, 44, 5, 1256-1261, 2016.11, Purpose: To evaluate the diagnostic performance of intravoxel incoherent motion (IVIM) MR imaging and 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) in differentiating primary central nervous system lymphoma (PCNSL) from glioblastoma multiforme (GBM). Materials and Methods: Fifty patients, 17 with PCNSL and 33 with GBM, were retrospectively studied. From the 3 Tesla IVIM data, the perfusion fraction (f) and diffusion coefficient (D) were obtained. In addition, the maximum standard uptake value (SUVmax) was obtained from the FDG-PET data. Each of the three parameters was compared between PCNSL and GBM using Mann-Whitney U-test. The performance in discriminating between PCNSL and GBM was evaluated using receiver-operating characteristics analysis and area-under-the-curve (AUC) values for the three parameters. Results: The fmax and Dmin values were significantly higher in GBM than in PCNSL (P < 0.01 and P < 0.0001, respectively). In addition, the SUVmax value was significantly lower in GBM than in PCNSL (P < 0.0005). The AUC values for fmax, Dmin, and SUVmax were 0.756, 0.905, and 0.857, respectively. The combination of the fmax and Dmin increased the diagnostic performance (AUC = 0.936) of fmax (P < 0.05), but this value was not significantly different from the values for Dmin (P = 0.30). Conclusion: IVIM-MR imaging noninvasively provides useful quantitative information in distinguishing between PCNSL and GBM. J. Magn. Reson. Imaging 2016;44:1256–1261..
25. Ryusuke Hatae, Nobuhiro Hata, Koji Yoshimoto, Daisuke Kuga, Yojiro Akagi, Hideki Murata, Satoshi O. Suzuki, Masahiro Mizoguchi, Koji Iihara, Precise detection of IDH1/2 and BRAF hotspot mutations in clinical glioma tissues by a differential calculus analysis of high-resolution melting data, PloS one, 10.1371/journal.pone.0160489, 11, 8, 2016.08, High resolution melting (HRM) is a simple and rapid method for screening mutations. It offers various advantages for clinical diagnostic applications. Conventional HRM analysis often yields equivocal results, especially for surgically obtained tissues. We attempted to improve HRM analyses for more effective applications to clinical diagnostics. HRM analyses were performed for IDH1R132 and IDH2R172 mutations in 192 clinical glioma samples in duplicate and these results were compared with sequencing results. BRAFV600E mutations were analyzed in 52 additional brain tumor samples. The melting profiles were used for differential calculus analyses. Negative second derivative plots revealed additional peaks derived from heteroduplexes in PCR products that contained mutations; this enabled unequivocal visual discrimination of the mutations. We further developed a numerical expression, the HRM-mutation index (MI), to quantify the heteroduplex-derived peak of the mutational curves. Using this expression, all IDH1 mutation statuses matched those ascertained by sequencing, with the exception of three samples. These discordant results were all derived from the misinterpretation of sequencing data. The effectiveness of our approach was further validated by analyses of IDH2R172 and BRAFV600E mutations. The present analytical method enabled an unequivocal and objective HRM analysis and is suitable for reliable mutation scanning in surgically obtained glioma tissues. This approach could facilitate molecular diagnostics in clinical environments..
26. Koji Yoshimoto, Akiko Kada, Daisuke Kuga, Ryusuke Hatae, Hideki Murata, Yojiro Akagi, Kunihiro Nishimura, Ryota Kurogi, Ataru Nishimura, Nobuhiro Hata, Masahiro Mizoguchi, Tetsuro Sayama, Koji Iihara, Current trends and healthcare resource usage in the hospital treatment of primary malignant brain tumor in Japan
A national survey using the diagnostic procedure combination database (J-ASPECT study-brain tumor), Neurologia medico-chirurgica, 10.2176/nmc.oa.2016-0172, 56, 11, 664-673, 2016, We conducted this study to clarify the current trends and healthcare resource usage in the treatment of inpatients with primary malignant brain tumors. The Diagnostic Procedure Combination (DPC) data of all inpatients treated between 2013 and 2014 in the 370 core and branch hospitals enrolled in the Japanese Neurosurgical Society training program were collected. DPC is a discharge abstract and administrative claims database of inpatients. We assessed 6,142 primary, malignant brain tumor patients. Patient information, diagnostic information, treatment procedure, and healthcare resource usage were analyzed. Chemotherapy was the most frequent treatment (27% of cases), followed by surgery (13%) and surgery + chemo-radiotherapy (11%). Temozolomide (TMZ), the most frequently used chemotherapeutic drug, was administered to 1,236 patients. Concomitant TMZ and radiotherapy was administered to 816 patients, and was performed according to the Stupp regimen in many cases. The mean length of hospital stay (LOS) was 16 days, and the mean medical cost was 1,077,690 yen. The average medical cost of TMZ-only treatment was 1,138,620 yen whilst it was 4,424,300 yen in concomitant TMZ patients. The LOS was significantly shorter in high-volume than in low-volume hospitals, and the medical cost was higher in hospitals treating 21–50 patients compared to those treating 1–10 patients. However, the direct medical cost of TMZ treatment was the same across different volume hospitals. This is the first report of current trends and healthcare resource usage in the treatment of primary malignant brain tumor inpatients in the TMZ era in Japan..
27. Hideki Murata, Koji Yoshimoto, Ryusuke Hatae, Yojiro Akagi, Masahiro Mizoguchi, Nobuhiro Hata, Daisuke Kuga, Akira Nakamizo, Toshiyuki Amano, Tetsuro Sayama, Koji Iihara, Detection of proneural/mesenchymal marker expression in glioblastoma
temporospatial dynamics and association with chromatin-modifying gene expression, Journal of Neuro-Oncology, 10.1007/s11060-015-1886-y, 125, 1, 33-41, 2015.10, Proneural and mesenchymal are two subtypes of glioblastoma identified by gene expression profiling. In this study, the primary aim was to detect markers to develop a clinically applicable method for distinguishing proneural and mesenchymal glioblastoma. The secondary aims were to investigate the temporospatial dynamics of these markers and to explore the association between these markers and the expression of chromatin-modifying genes. One hundred thirty-three glioma samples (grade II: 14 samples, grade III: 18, grade IV: 101) were analyzed. We quantified the expression of 6 signature genes associated with proneural and mesenchymal glioblastoma by quantitative reverse transcription-polymerase chain reaction. We assigned proneural (PN) and mesenchymal (MES) scores based on the average of the 6 markers and calculated a predominant metagene (P-M) score by subtracting the MES from the PN score. We used these scores to analyze correlations with malignant transformation, tumor recurrence, tumor heterogeneity, chromatin-modifying gene expression, and HDAC7 expression. The MES score positively correlated with tumor grade, whereas the PN score did not. The P-M score was able to distinguish the proneural and mesenchymal subtypes. It was decreased in cases of tumor recurrence and malignant transformation and showed variability within a tumor, suggesting intratumoral heterogeneity. The PN score correlated with the expression of multiple histone-modifying genes, whereas the MES score was associated only with HDAC7 expression. Thus, we demonstrated a simple and straightforward method of quantifying proneural/mesenchymal markers in glioblastoma. Of note, HDAC7 expression might be a novel therapeutic target in glioblastoma treatment..
28. Xinlong Ma, Koji Yoshimoto, Yaulei Guan, Nobuhiro Hata, Masahiro Mizoguchi, Noriaki Sagata, Hideki Murata, Daisuke Kuga, Toshiyuki Amano, Akira Nakamizo, Tomio Sasaki, Associations between microRNA expression and mesenchymal marker gene expression in glioblastoma, Neuro-Oncology, 10.1093/neuonc/nos145, 14, 9, 1153-1162, 2012.09, The subclassification of glioblastoma (GBM) into clinically relevant subtypes using microRNA (miRNA)- and messenger RNA (mRNA)-based integrated analysis has been attempted. Because miRNAs regulate multiple gene-signaling pathways, understanding miRNA-mRNA interactions is a prerequisite for understanding glioma biology. However, such associations have not been thoroughly examined using high-throughput integrated analysis. To identify significant miRNA-mRNA correlations, we selected and quantified signature miRNAs and mRNAs in 82 gliomas (grade II: 14, III: 16, IV: 52) using real-time reverse-transcriptase polymerase chain reaction. Quantitative expression data were integrated into a single analysis platform that evaluated the expression relationship between miRNAs and mRNAs. The 21 miRNAs include miR-15b, -21, -34a, -105, -124a, -128a, -135b, -184, -196a-b, -200a-c, -203, -302a-d, -363, -367, and -504. In addition, we examined 23 genes, including proneural markers (DLL3, BCAN, and OLIG2), mesenchymal markers (YKL-40, CD44, and Vimentin), cancer stem cell-related markers, and receptor tyrosine kinase genes. Primary GBM was characterized exclusively by upregulation of mesenchymal markers, whereas secondary GBM was characterized by significant downregulation of mesenchymal markers, miR-21, and -34a, and by upregulation of proneural markers and miR-504. Statistical analysis showed that expression of miR-128a, -504, -124a, and -184 each negatively correlated with the expression of mesenchymal markers in GBM. Our functional analysis of miR-128a and -504 as inhibitors demonstrated that suppression of miR-128a and -504 increased the expression of mesenchymal markers in glioblastoma cell lines. Mesenchymal signaling in GBM may be negatively regulated by miR-128a and -504..
29. Koji Yoshimoto, Xinlong Ma, Yaulei Guan, Masahiro Mizoguchi, Akira Nakamizo, Toshiyuki Amano, Nobuhiro Hata, Daisuke Kuga, Tomio Sasaki, Expression of stem cell marker and receptor kinase genes in glioblastoma tissue quantified by real-time RT-PCR, Brain tumor pathology, 10.1007/s10014-011-0046-0, 28, 4, 291-296, 2011.10, Glioblastoma is dependent on a specific signaling pathway to maintain its tumor phenotype. The receptor tyrosine kinase (RTK) family mediates the multiple oncogenic growth factor receptor signaling and contributes to the pathogenesis of glioblastoma. Recently, many studies have shown that the expression of stem cell marker in glioblastoma tissue has prognostic significance, which indicates that the quantification of stem cell markers and RTK genes yields biological information about glioblastoma. In this study, we quantified RNA expression levels of stem cell markers [CD133, Nestin, BMI-1, maternal embryonic leucine zipper kinase (MELK), and Notch1-4] as well as RTKs (EGFR, ErbB4, VEGFR1-3, FGFR1, -2, PDGFRA, and PDGFRB) in 42 clinical samples of glioblastoma by the real-time RT-PCR method. We demonstrated that the expression of MELK is exclusively upregulated in glioblastoma tissue. Notch receptor expression is moderately upregulated and is correlated with that of VEGFR2, VEGFR3, and PDGFR β. Unsupervised clustering identified one unique sample group that showed high expression of most of the genes analyzed. Our results suggest that quantification of these stem cell markers and RTK genes can stratify patients based on the expression profile, which might provide insight into the glioma biology in each cluster..
30. Masahiro Mizoguchi, Daisuke Kuga, Yanlei Guan, Nobuhiro Hata, Akira Nakamizo, Koji Yoshimoto, Tomio Sasaki, Erratum
Loss of heterozygosity analysis in malignant gliomas (Brain Tumor Pathol DOI: 10.1007/s10014-011-0038-0), Brain tumor pathology, 10.1007/s10014-011-0049-x, 28, 3, 197, 2011.07.
31. Masahiro Mizoguchi, Daisuke Kuga, Yanlei Guan, Nobuhiro Hata, Akira Nakamizo, Koji Yoshimoto, Tomio Sasaki, Loss of heterozygosity analysis in malignant gliomas, Brain tumor pathology, 10.1007/s10014-011-0038-0, 28, 3, 191-196, 2011.07, Despite recent advances in the diagnosis and treatment of glioblastomas, patient outcomes for these highly malignant tumors remain poor. Research into the molecular pathology of glioblastoma has uncovered various genetic changes that contribute to malignancy. Some of the identified molecular markers-such as loss of heterozygosity (LOH) on chromosome 1p/19q and chromosome 10, O6-methylguanine methyltransferase promoter hypermethylation, and mutation of isocitrate dehydrogenase-1-may help to predict patient outcomes. Indeed, LOH analysis is an effective approach to classify malignant gliomas. Genome-wide analyses have revealed that the extent and pattern of LOH regions may have important implications for the clinical course of the disease. As the genetic underpinnings of malignant gliomas are complex and varied, careful selection of the methods for genetic analysis in the clinic is important. The fundamental principles of each assay need to be understood to allow careful selection of practically useful methods. This review summarizes recent developments in the molecular analysis of malignant glioma..
32. Yanlei Guan, Masahiro Mizoguchi, Koji Yoshimoto, Nobuhiro Hata, Tadahisa Shono, Satoshi O. Suzuki, Yukie Araki, Daisuke Kuga, Akira Nakamizo, Toshiyuki Amano, Xinlong Ma, Kenshi Hayashi, Tomio Sasaki, MiRNA-196 is upregulated in glioblastoma but not in anaplastic astrocytoma and has prognostic significance, Clinical Cancer Research, 10.1158/1078-0432.CCR-10-0207, 16, 16, 4289-4297, 2010.08, Purpose: MicroRNAs (miRNA) are short noncoding RNAs that can play critical roles in diverse biological processes. They are implicated in tumorigenesis and function both as tumor suppressors and as oncogenes. The clinical significance of miRNA expression profiles in malignant gliomas remains unclear. Experimental Design: In this study, we examined the expression levels of 365 mature human miRNAs in 12 malignant gliomas, including 8 glioblastomas and 4 anaplastic astrocytomas, using TaqMan real-time quantitative PCR arrays. A validation study was done to corroborate a subset of the results, including expression levels of miR-196a, -196b, -21, and -15b, by analyzing 92 malignant gliomas by conventional real-time PCR. We modeled the relationship between the expression levels of these miRNAs and the survival rate of 39 glioblastoma patients by Kaplan-Meier method and multivariate analysis. Results: Expression profiles in glioblastomas and anaplastic astrocytomas suggested that 16 miRNAs were candidate markers associated with the malignant progression of gliomas. Among them, miR-196a showed the most significant difference (P = 0.0038), with miR-196b also having a high significance (P = 0.0371). Both miRNAs showed increased expression levels in glioblastomas relative to both anaplastic astrocytomas and normal brains in the validation study. Furthermore, patients with high miR-196 expression levels showed significantly poorer survival by the Kaplan-Meier method (P = 0.0073). Multivariate analysis showed that miR-196 expression levels were an independent predictor of overall survival in all 39 glioblastoma patients (P = 0.021; hazard ratio, 2.81). Conclusions: Our results suggest that miR-196 may play a role in the malignant progression of gliomas and may be a prognostic predictor in glioblastomas..