Atsushi Kambe

Proteasome inhibitor MG132 induces NAG-1/GDF15 expression through the p38 MAPK pathway in glioblastoma cells.

The expression of nonsteroidal anti-inflammatory drug-activated gene-1 (NAG-1) is regulated by the p53 and Egr-1 tumor suppressor pathways. Many anti-cancer drugs and chemicals induce NAG-1 expression, but the mechanisms are not fully understood. Transgenic mice expressing human NAG-1 are resistant to intestinal and prostate cancer, suggesting that NAG-1 is a tumor suppressor. Proteasome inhibitors exhibit anti-glioblastoma activities in preclinical studies. Here, we show that the proteasome inhibitors MG132 and bortezomib induced NAG-1 expression and secretion in glioblastoma cells. MG132 increased NAG-1 expression through transcriptional and post-transcriptional mechanisms. At the transcriptional level, the induction of NAG-1 required the -133 to +41 bp region of the promoter. At post-transcriptional levels, MG132 stabilized NAG-1 mRNA by increasing the half-life from 1.5 h to >8 h. Because of the dramatic increase in mRNA stability, this is likely the major contributor to MG132-mediated NAG-1 induction. Further probing into the mechanism revealed that MG132 increased phosphorylation of the p38 MAPK pathway. Consequently, inhibiting p38 phosphorylation blocked activation of the NAG-1 promoter and decreased mRNA stability, indicating that p38 MAPK activation mediates both MG132-dependent promoter activation and mRNA stabilization of NAG-1. We propose that the induction of NAG-1 by p38 MAPK is a potential contributor to the anti-glioblastoma activity of proteasome inhibitors.

Spatial resolution of calpain-catalyzed proteolysis in focal cerebral ischemia.

Transient forebrain ischemia induces calpain-mediated degradation of the neuronal cytoskeleton, alpha-fodrin, and this results in ischemic neuronal death. In this study, we investigated the spatial distribution and temporal changes of calpain-catalyzed alpha-fodrin proteolysis in focal cerebral ischemia and examined the effects of a calpain inhibitor. Ischemia was induced in gerbils by 3-h middle cerebral artery occlusion followed by reperfusion. Animals were divided into four groups: a sham-operated group, an ischemic group, a vehicle-treated group, and a calpain inhibitor-treated group. Intravenous injections of vehicle or calpain inhibitor I were administered 30 min before ischemia. Infarct volumes were measured 1 day after reperfusion and the spatial distribution of calpain-catalyzed alpha-fodrin proteolysis was investigated by immunohistochemistry 15 min, 1 h, 4 h, and 1 day after reperfusion. Infarct volume (mean +/- SD) in the ischemic group and the vehicle-treated group was 204.6 +/- 19.1 mm3 and 212.4 +/- 16.3 mm3, respectively, and the calpain inhibitor I reduced the infarct volume [149.4 +/- 25.2 mm3 (P < 0.05)]. Immunoblot analysis demonstrated that calpain inhibitor reduced proteolysis. Ischemia induced fodrin proteolysis in the ischemic core and the peri-infarct zone within 15 min after reperfusion, with proteolysis developing quickly in the ischemic core and more slowly in the peri-infarct zone. Proteolysis preceded neuronal death in the peri-infarct zone. Calpain inhibitor I ameliorated neuronal death in the peri-infarct zone but not in the ischemic core. Thus, calpain plays a pivotal role on focal ischemia as well as in global ischemia.

The Cyclooxygenase Inhibitor Sulindac Sulfide Inhibits EP4 Expression and Suppresses the Growth of Glioblastoma Cells

EP4 expression in human glioblastoma cells correlates with growth on soft agar. The cyclooxygenase inhibitor sulindac sulfide first altered specificity protein-1 (Sp-1) and early growth response gene-1 expression, then increased the expression of nonsteroidal anti-inflammatory drug-activated gene 1 and activating transcription factor 3, and then decreased EP4 expression. EP4 suppression was dependent on blocking the Sp-1 binding sites in the human EP4 promoter. Mutation in the Sp-1 sites in EP4 altered the promoter activity and abolished sulindac sulfide effects. The inhibitory effect of sulindac sulfide on EP4 expression was reversed by PD98059, a mitogen-activated protein/extracellular signal–regulated kinase kinase-1/extracellular signal–regulated kinase inhibitor. Sp-1 phosphorylation was dependent on sulindac sulfide–induced Erk activation. Chromatin immunoprecipitation assay confirmed that Sp-1 phosphorylation decreases Sp-1 binding to DNA and leads to the suppression of EP4. Inhibition of cell growth on soft agar assay was found to be a highly complex process and seems to require not only the inhibition of cyclooxygenase activity but also increased expression of nonsteroidal anti-inflammatory drug-activated gene 1 and activating transcription factor 3 and suppression of EP4 expression. Our data suggest that the suppression of EP4 expression by sulindac sulfide represents a new mechanism for understanding the tumor suppressor activity.

Intramedullary recurrence of germinoma in the spinal cord 15 years after complete remission of a pineal lesion

The authors present a case of germinoma that was initially found in the pineal region and recurred 15 years later in the intramedullary cervical spinal cord after intensive chemo- and radiotherapy and diagnosis of complete remission. This 28-year-old man initially presented with seizures. Hydrocephalus and a pineal tumor were found on radiological examination, and partial resection of the tumor was performed. Histological diagnosis showed a pure germinoma. Following surgery, the patient received a combination of chemo- and radiotherapy, and a complete remission was shown. However, after 15 years of follow-up, he presented with gait disturbances. Spinal MRI showed an intramedullary mass lesion in the cervical spinal cord. The cervical lesion was biopsied, and histological examination again revealed a pure germinoma. With germinomas, the possibility of a drop metastasis from an intracranial lesion to the spinal cord must be considered during follow-up. However, in the present case, analysis of a CSF sample showed no abnormalities as in previously published cases. In recent years, multidisciplinary treatments have demonstrated good event-free survival rates in cases of pure germinomas, but long-term outcomes over the decades are not fully known. Continual follow-up of such cases is recommended even after complete remission has been achieved.

Serial 3 T magnetic resonance imaging during cabergoline treatment of macroprolactinomas

Abstract Objective: Cabergoline is the treatment of choice for prolactin (PRL)-producing pituitary adenomas, because of its efficacy in normalizing PRL levels, and inducing tumor shrinkage. The clinical use of 3 T magnetic resonance imaging (MRI) for neuroimaging has rapidly expanded in recent years. In particular, T2-weighted imaging (T2WI) provides high anatomical and contrast resolution. Patients and methods: In this study, serial 3 T MRI with T2WI was utilized during cabergoline treatment of 10 patients with macroprolactinomas. Cabergoline was started at a standard weekly dosage and incrementally adjusted on individual posttreatment PRL values. Results: MRI confirmed tumor shrinkage in all patients during cabergoline treatment. Cabergoline normalized hyperprolactinemia in all but one patient. In six of 10 patients, distinct low-signal-intensity areas were evident throughout the adenomas on T2WI. In four of those six patients, massive low-signal-intensity areas appeared at 1–4 months, after which tumors decreased in size by over 80%. These findings in the early phase of prolactinoma treatment predicted pronounced regression or near-complete disappearance of the tumor. Reduction of T2 intensity possibly reflected dehydration due to diffuse hemorrhage in the adenomas. Conclusion: T2-weighted 3 T MR images are valuable for assessing and monitoring cabergoline treatment of macroprolactinomas.

Pediatric Case of Li-Fraumeni Syndrome Complicated with Supratentorial Anaplastic Ependymoma

BACKGROUND Li-Fraumeni syndrome is a genetic disease that is caused by mutation of the tumor suppressor gene TP53. Patients with this syndrome may develop multiple malignant neoplasms including brain tumors. We herein report the first case of Li-Fraumeni syndrome in which development of supratentorial anaplastic ependymoma led to difficulty in terms of selecting the optimal postoperative therapeutic protocol. CASE DESCRIPTION A 7-year-old boy experiencing a convulsive attack was brought to our institute. He underwent surgical tumor resection, and magnetic resonance imaging of the head revealed a tumor-like lesion in the right parietal lobe. Although adjuvant radiotherapy was performed after total tumor resection, a focal recurrent lesion appeared soon afterward. We initiated chemotherapy with bevacizumab after resection of the recurrent lesion, but bevacizumab was unable to control tumor progression. At this writing, he remains bedridden and requires tube feeding and artificial ventilation. CONCLUSION Since Li-Fraumeni syndrome is a genetic disease that is caused by mutation of the tumor suppression gene TP53, patients should generally not be treated with radiotherapy or alkylating agents that induce deoxyribonucleic acid damage. However, if the prognostic benefit of postoperative adjuvant therapies is thought to surpass the risk of long-term secondary cancer, it is appropriate to consider these therapies after consultation with the patient and family. Postoperative treatment protocols are controversial, and their role should be further explored in cases of Li-Fraumeni syndrome complicated with malignant gliomas.

Superficial Siderosis Associated with Pineal Cavernous Malformation

BACKGROUND Cavernous malformations in the pineal region are rare and difficult to anticipate from preoperative evaluation in patients with pineal apoplexy. We herein report the first case of a pineal cavernous malformation with superficial siderosis. Radiological findings were helpful in identifying the presence of the cavernous malformation. CASE DESCRIPTION A 47-year-old female presented with a 4-month history of progressive headache, nausea, and dizziness. She complained of double vision and exhibited upward gaze palsy and papilledema on fundoscopy. Radiological examination revealed subacute hemorrhage in the pineal region and enlarged lateral ventricles. Furthermore, T2-star-weighted gradient-echo magnetic resonance imaging demonstrated a linear hypointensity along the pial surface of the cerebral cortex, brainstem, and cerebellum, indicating hemosiderin deposition consistent with superficial siderosis. Suspecting the presence of a cavernous malformation based on the radiological findings of superficial siderosis, we performed total mass resection. The postoperative course was uneventful and her preoperative symptoms resolved completely. CONCLUSION Radiological findings of superficial siderosis on T2-star-weighted gradient-echo imaging are useful to making a diagnosis of cavernous malformation in cases of pineal apoplexy. They are also important for making the treatment decision to perform total mass resection, which is the best curative method for pineal cavernous malformations.

Utility of intravoxel incoherent motion magnetic resonance imaging and arterial spin labeling for recurrent glioma after bevacizumab treatment

Background Detecting recurrence of glioma on magnetic resonance imaging (MRI) is getting more and more important, especially after administration of new anti-tumor agent. However, it is still hard to identify. Purpose To examine the utility of intravoxel incoherent motion (IVIM) MRI and arterial spin labeling-cerebral blood flow (ASL-CBF) for recurrent glioma after initiation of bevacizumab (BEV) treatment. Material and Methods Thirteen patients (7 men, 6 women; age range = 41–82 years) with glioma (high grade, n = 11; low grade, n = 2) were enrolled in the study. IVIM parameters including apparent diffusion coefficient (ADC), true diffusion coefficient (D), and perfusion fraction (f) were obtained with 14 different b-values. We identified tumor progression during BEV therapy by MRI monitoring consisting of diffusion-weighted imaging (DWI), fluid-attenuated inversion recovery (FLAIR) imaging, and contrast-enhanced T1-weighted (CE-T1W) imaging by measuring tumor area. We also measured each parameter of IVIM and ASL-CBF, and calculated relative ADC (rADC), relative D (rD), relative f (rf), and relative CBF (rCBF) by obtaining the ratio between each area and the contralateral cerebral white matter. We calculated the rate of change (Δ) by subtracting values from those from the preceding MRI study, and obtained Spearman’s rank correlation coefficient (rs). Results Tumor progression was identified in nine patients (high grade, n = 7; low grade, n = 2). Negative correlations were identified between ΔrD and ΔDWI area (rs = –0.583), and between ΔrD and ΔCE-T1W imaging area (rs = –0.605). Conclusion Tumor progression after BEV treatment can be identified by decreasing rD.