Masayuki Kanamori

Analysis of IDH1 and IDH2 mutations in Japanese glioma patients

A recent study reported on mutations in the active site of the isocitrate dehydrogenase 1 (IDH1) gene in several types of gliomas. All mutations detected resulted in an amino acid exchange at position 132. We analyzed the genomic region spanning wild‐type R132 of IDH1 by direct sequencing in 125 glial tumors. A total of 39 IDH1 mutations were observed. Mutations of the IDH2 gene, homologous to IDH1, were often detected in gliomas without IDH1 mutations. In the present study, R172 mutation of the IDH2 gene was detected in one anaplastic astrocytoma. IDH1 or IDH2 mutations were frequently in oligodendrogliomas (67%), anaplastic astrocytomas (62%), anaplastic oligoastrocytomas (75%), anaplastic oligodendrogliomas (50%), secondary glioblastomas (67%), gangliogliomas (38%), and anaplastic gangliogliomas (60%). Primary glioblastomas were characterized by a low frequency of mutations (5%) at amino acid position 132 of IDH1. Mutations of the IDH1 or IDH2 genes were significantly associated with improved outcome in patients with anaplastic astrocytomas. Our data suggest that IDH1 or IDH2 mutation plays a role in early tumor progression of several types of glioma and might arise from a common glial precursor. The infrequency of IDH1 mutation in primary glioblastomas revealed that these subtypes are genetically distinct entities from other glial tumors. (Cancer Sci 2009; 100: 1996–1998)

Integrin β3 Overexpression Suppresses Tumor Growth in a Human Model of Gliomagenesis Implications for the Role of β3 Overexpression in Glioblastoma Multiforme

alphaVbeta3 integrin complexes are overexpressed in the growing, invading margins of human glioblastoma multiforme (GBM) and in the GBM vasculature, suggesting a key role for alphaVbeta3 in GBM growth and invasion. The function of alphaVbeta3 complexes in tumor formation, however, has been challenged by studies showing that loss of alphaVbeta3 expression (via loss of beta3) in the host vasculature enhances, rather than suppresses, the growth of s.c. implanted carcinomas. To directly address the role of tumor-specific alphaVbeta3 overexpression in glioma formation, we increased alphaVbeta3 expression (via overexpression of a wild-type or constitutively activated beta3) in human astrocytes genetically modified to form anaplastic astrocytoma-like tumors (Ras cells) on intracranial injection in rats. Overexpression of beta3 selectively increased levels of alphaVbeta3 integrin complexes, but had no effect on anchorage-dependent or -independent growth in vitro. After intracranial injection, however, the Ras + beta3 cells formed fewer and smaller tumors than did Ras cells. Similarly, Ras-transformed mouse astrocytes that were derived from control animals formed smaller intracranial tumors than those derived from beta3 knockout animals. Although tumors formed by human Ras and Ras + beta3 cells were similar in blood vessel density, Ras + beta3 tumors had smaller, pericyte-depleted vessels and were significantly more hypoxic, suggesting a beta3-mediated vascular defect. The growth-suppressive actions of beta3, however, could be overcome by stimulation of pathways (Akt or vascular endothelial growth factor) commonly activated in GBM. These results show that tumor-specific alphaVbeta3 overexpression has growth-suppressive effects in gliomas, but that these deleterious effects are mitigated by alterations common to alphaVbeta3-overexpressing GBM.

The PTEN/Akt Pathway Dictates the Direct αVβ3-Dependent Growth-Inhibitory Action of an Active Fragment of Tumstatin in Glioma Cells In vitro and In vivo

The collagen type IV cleavage fragment tumstatin and its active subfragments bind to integrin alpha(V)beta(3) and inhibit activation of focal adhesion kinase, phophoinositol-3 kinase, Akt, and mammalian target of rapamycin (mTOR) in what is thought to be an endothelial cell-specific manner. The resultant endothelial cell apoptosis accounts for the ability of tumstatin to function as an endogenous inhibitor of angiogenesis and an indirect suppressor of tumor growth. We hypothesized that the inability of tumstatin to directly suppress tumor cell growth might be the result of the constitutive activation of the Akt/mTOR pathway commonly seen in tumors. Consistent with this idea, several integrin alpha(V)beta(3)-expressing glioma cell lines with PTEN mutations and high levels of phospho-Akt (pAkt) were unaffected by exposure to an active fragment of tumstatin (T3), whereas alpha(V)beta(3)-expressing glioma cell lines with a functional PTEN/low levels of pAkt exhibited T3-induced growth suppression that could be bypassed by small interfering RNA-mediated suppression of PTEN, introduction of a constitutively expressed Akt, or introduction of the Akt and mTOR target eukaryotic translation initiation factor 4E. The direct tumor-suppressive actions of T3 were further shown in an alpha(V)beta(3)-deficient in vivo mouse model in which T3, while unable to alter the tumstatin-insensitive vasculature contributed by the alpha(V)beta(3)-deficient host, nonetheless suppressed the growth and proliferative index of i.c. implanted alpha(V)beta(3)-expressing PTEN-proficient glioma cells. These results show that tumstatin, previously considered to be only an endogenous inhibitor of angiogenesis, also directly inhibits the growth of tumors in a manner dependent on Akt/mTOR activation.

Distinctive flow pattern of wall shear stress and oscillatory shear index: similarity and dissimilarity in ruptured and unruptured cerebral aneurysm blebs

OBJECT The difference in the hemodynamics of wall shear stress (WSS) and oscillatory shear index (OSI) between ruptured and unruptured aneurysms is not well understood. The authors investigated the hemodynamic similarities and dissimilarities in ruptured and thin-walled unruptured aneurysm blebs. METHODS Magnetic resonance imaging-based fluid dynamics analysis was used to calculate WSS and OSI, and hemodynamic and intraoperative findings were compared. The authors also compared ruptured and unruptured thin-walled blebs for the magnitude of WSS and OSI. RESULTS Intraoperatively, 13 ruptured and 139 thin-walled unruptured aneurysm blebs were identified. Twelve of the ruptured (92.3%) and 124 of the unruptured blebs (89.2%) manifested low WSS and high OSI. The degree of WSS was significantly lower in ruptured (0.49 ± 0.12 Pa) than in unruptured (0.64 ± 0.15 Pa; p < 0.01) blebs. CONCLUSIONS Ruptured and unruptured blebs shared a distinctive pattern of low WSS and high OSI. The degree of WSS at the rupture site was significantly lower than in the unruptured thin-walled blebs.

Microsatellite instability and the PTEN1 gene mutation in a subset of early onset gliomas carrying germline mutation or promoter methylation of the hMLH1 gene.

High-frequent microsatellite instability (MSI-H) was detected in two of the 80 gliomas examined, whlie the other 78 gliomas showed microsatellite stable (MSS) phenotype. Both of the two MSI-H tumors were glioblastomas which developed in teenage patients. One of the patient was diagnosed as having Turcots syndrome and had a germline mutation in the hMLH1 gene. Loss of expression due to promoter methylation was selectively observed in the wild type allele of the hMLH1 gene in the tumor of this patient. The other patient had neither a family history nor a past personal history of malignancy. Although no mutation in the mismatch repair genes was detected in the tumor of this patient, the level of expression of the hMLH1 gene was markedly decreased and the promoter sequence of the gene was highly methylated. In the tumor of this patient, the PTEN1 gene, one of the genes carrying microsatellite sequences in their coding regions, was altered by a slippage mutation within five adenine repeat sequences. These findings indicate that the genetic or epigenentic inactivation of the hMLH1 gene is involved in a subset of early-onset gliomas and the PTEN1 gene could be a downstream target for mutation as observed in glioblastoma without MSI.

Intracranial microenvironment reveals independent opposing functions of host αvβ3 expression on glioma growth and angiogenesis

αVβ3 integrins are overexpressed in the host-derived vasculature of glioblastoma multiform (GBM) and are believed to contribute to angiogenesis and tumor growth. To directly address the role of host αVβ3 expression in GBM growth and behavior, we intracranially implanted integrin β3-expressing GBM cells into β3 wild type (WT) or β3 knock out (KO) mice and monitored angiogenesis and growth. GBM in β3 WT animals had a vessel density greater than that in β3 KO animals, consistent with a pro-angiogenic, pro-tumorigenic view of host integrin function. GBM in β3 WT animals, however, were no larger than those in β3 KO animals, because GBM in β3WT animals were infiltrated with a higher number of tumor necrosis factor α-secreting, apoptosis-inducing macrophages than the tumors in the corresponding β3 KO animals. The tumor-suppressive effects of host β3 expression could be reversed by macrophage depletion or by transplantation of bone marrow from β3 KO animals into β3 WT animals, both of which significantly increased tumor growth independently of tumor vessel density. Taken together, these results show that host αVβ3 integrin expression has opposing actions in the intracranial setting, enhancing tumor vascularization and growth while independently enhancing macrophage-mediated tumor elimination. Appropriate management of these functions could lead to enhanced efficacy of anti-integrin based therapies for glioma.

Papillary tumor of the pineal region: a case report.

We report a case of papillary tumor of the pineal region (PTPR) and describe the morphological, immunohistochemical, and neuroimaging findings. A 43-year-old man presented with signs of increased intracranial pressure and upward gaze palsy. Magnetic resonance (MR) imaging demonstrated a heterogeneously enhanced mass in the pineal region and obstructive hydrocephalus. Proton MR spectroscopy revealed increased choline and decreased N-acetyl aspartate peaks with a slightly increased lactate peak. Minimum apparent diffusion coefficient value was 0.60 × 10−3 mm2/s. Positron emission tomography showed significantly increased [18F]fluorodeoxyglucose uptake at the site of the lesion. He underwent total resection of the pineal region mass, resulting in resolution of the symptoms. The tumor consisted of columnar and cuboidal cells, with papillary growth pattern. Immunohistochemical staining showed positive reaction for neuron-specific enolase, S-100 protein, and vimentin. Ki-67 labeling index (LI) was 13.1%. These features were consistent with PTPR. Postoperatively, the patient received radiochemotherapy, and maintenance chemotherapy at our outpatient clinic, and was doing well without tumor recurrence 1 year after the surgery. Although the morphological features agree with those in the original description of the PTPR, the Ki-67 LI and radiologic findings suggest the malignant nature and the necessity for adjuvant therapy.

Germ cell tumors in the basal ganglia: problems of early diagnosis and treatment

OBJECT Intracranial germ cell tumors (GCTs) originating in the basal ganglia are rare. The authors investigated factors related to the diagnosis of these lesions as well as outcome in order to help decrease the time to diagnosis and improve treatment efficacy. METHODS The authors reviewed the clinical features of 142 cases of intracranial GCT in their institute. Fourteen cases of basal ganglia GCT were identified. The symptoms, neuroimaging findings, delay between symptom onset and diagnosis or treatment, initial and further treatment, and outcome were investigated. RESULTS Major symptoms were motor weakness and precocious puberty. Gadolinium-enhanced T1-weighted MR images showed enhancement in 8 of 11 patients examined, but only slight hyperintensity without enhancement in 2 patients. Ipsilateral peduncle and hemispheric atrophy were found in 3 and 4 patients, respectively. Cases of basal ganglia GCT were characterized by a longer delay from the initial neuroimaging examination to diagnosis compared with GCT in other regions. Five patients had aggravated hemiparesis in the extremities due to the delay in diagnosis. Despite good response to the initial therapy, 5 patients experienced recurrence; 2 of these 5 had malignant GCTs, and 3 had been treated only with chemotherapy or radiochemotherapy with insufficient radiation dose and field. Finally, the 2 patients with malignant GCTs died of the disease, and 1 died of aspiration pneumonia due to dissemination around the brainstem. CONCLUSIONS Early diagnosis requires MR imaging with administration of contrast medium in young patients presenting with motor weakness and/or precocious puberty. Serial neuroimaging studies should be performed if any tiny lesion is detected in the basal ganglia. Since insufficient treatment resulted in early recurrence, radiation therapy with adequate dose and field is essential.

The expression status of CD133 is associated with the pattern and timing of primary glioblastoma recurrence.

BACKGROUND Glioblastoma carries a poor prognosis primarily because of its high rate of recurrence. The ability to predict the recurrence pattern and timing would be highly useful for determining effective treatment strategies. We examined the correlation between prognostic factors and the pattern of recurrence in patients with primary glioblastoma. In particular, we examined whether there was a correlation between the expression of CD133 and glioblastoma recurrence. METHODS We retrospectively analyzed 112 patients with primary glioblastoma. The timing and pattern (local or distant) of the initial recurrence were obtained from medical records. To identify factors predictive of recurrence, we examined CD133 expression by Western blots and immunohistochemistry, clinical (age, sex, KPS, Ki67 labeling index, surgery, ventricular entry) and genetic (IDH1, 7p, 9p, 10q, MGMT) factors. RESULTS Of the 112 patients, 99 suffered recurrence. The first recurrence was local in 77 patients and distant in 22 patients. Among the factors to predict the pattern of recurrence, CD133 expression was significantly higher in distant than in local recurrence. Of the factors to predict the timing of recurrence, high CD133 expression was associated with shorter time to distant recurrence in both univariate and multivariate analyses (P = .0011 and P = .038, respectively). CONCLUSIONS The expression of CD133 may be a predictor of the pattern and timing of recurrence of primary glioblastoma.

201Tl-SPECT, 1H-MRS, and MIB-1 labeling index of central neurocytomas: Three case reports

Summary.Summary. Background: The proliferative activity and metabolic features of three central neurocytomas were investigated using the findings of thallium-201 single photon emission computed tomography (201Tl-SPECT) and proton magnetic resonance spectroscopy (1H-MRS), and the MIB-1 labeling index (MIB-1 LI). Method: The early and delayed 201Tl indixes were calculated as the ratio of tumour to normal brain tissue uptake by 201Tl-SPECT. In vivo single-voxel 1H-MRS was performed with echo time of 272 msec to evaluate the metabolites including choline (Cho), N-acetyl aspartate (NAA) and creatine/phosphocreatine (Cre). An external standard reference was used to semiquantitate each metabolite. MIB-1 LI was determined in the surgical specimens. Findings: The MIB-1 LI was 0.5%, 1.2%, and 7.5% in an atypical central neurocytoma without intraventricular extension. Significant 201Tl uptake was observed on delayed images in all three central neurocytomas. 1H-MRS showed the high Cho peaks relative to the NAA and Cre peak. The signal at 3.55 ppm, which may be due to inositol or glycine, was observed in one central neurocytoma. Interpretation. Both 201Tl-SPECT and 1H-MRS did not reflect the proliferative potential of central neurocytomas.