Critical role of platelet-derived growth factor-α in angiogenesis after indirect bypass in a murine moyamoya disease model.

OBJECTIVE: This study aimed to clarify the underlying mechanism of pathognomonic angiogenesis between the temporal muscle and neocortex after indirect bypass for moyamoya disease by shedding light on the role of platelet-derived growth factor receptor-α (PDGFRα) in angiogenesis. METHODS: The gene for PDGFRα was systemically inactivated in adult mice (α-KO mice). The Pdgfra-preserving mice (Flox mice) and α-KO mice were exposed to bilateral common carotid artery stenosis (BCAS) by using microcoils. One week later the animals underwent encephalomyosynangiosis (EMS) on the right side. Cerebral blood flow (CBF) was serially measured using a laser Doppler flowmeter. Histological analysis was performed on the distribution of CD31-positive vessels and collagen deposit at 28 days after BCAS. Reverse transcription polymerase chain reaction (RT-PCR) was performed to assess the expression of collagen mRNA in the skin fibroblasts derived from Flox and α-KO mice. RESULTS: BCAS significantly reduced CBF up to approximately 70% of the control level at 28 days after the onset. There was no significant difference in CBF between Flox and α-KO mice. EMS significantly enhanced the improvement of CBF on the ipsilateral side of Flox mice, but not α-KO mice. EMS significantly induced the development of CD31-positive vessels in both the neocortex and temporal muscle on the ipsilateral side of Flox mice, but not α-KO mice. Deposition of collagen was distinctly observed between them in Flox mice, but not α-KO mice. Expression of mRNA of collagen type 1 alpha 1 (Col1a1) and collagen type 3 alpha 1 (Col3a1) was significantly downregulated in the skin fibroblasts from α-KO mice. CONCLUSIONS: This is the first study that denotes the role of a specific growth factor in angiogenesis after EMS for moyamoya disease by inactivating its gene in mice. The findings strongly suggest that PDGFRα signal may play an important role in developing spontaneous angiogenesis between the temporal muscle and neocortex after EMS in moyamoya disease.

Novel biomarker, phosphorylated T-LAK cell-originated protein kinase (p-TOPK) can predict outcome in primary central nervous system lymphoma.

This study aimed to assess whether T-lymphokine-activated killer cell-originated protein kinase (TOPK) can be a potent novel biomarker to predict the outcome in patients with primary central nervous system lymphoma (PCNSL). This study enrolled 20 patients who were histologically diagnosed as having diffuse large B-cell type PCNSL between 2005 and 2015. Using surgical specimens, the expression of TOPK and phosphorylated TOPK (p-TOPK) was analyzed on immunohistochemistry. Clinical features such as age, sex, Karnofsky performance status (KPS), ocular involvement, deep brain structure involvement, the number of lesions, chemotherapy and radiation therapy were also collected. Impacts of TOPK/p-TOPK expression on their progression-free survival (PFS) and overall survival (OS) were examined with multivariate analysis. Median PFS/OS were 24.2 and 39.0 months, respectively. On immunostaining, the mean percentage of TOPK-positive cells was 35.5 ± 20.8%, and the mean number of p-TOPK-positive cells was 13.7 ± 15.7 cells/mm2 . The higher expression of p-TOPK was significantly related to multiple lesions (P = 0.003). Multivariate analysis demonstrated that only the higher expression of p-TOPK was an independent predictor to shorten both PFS (P = 0.029; hazard ratio (HR), 5.5; 95% confidential interval (CI), 1.2-25.3) and OS (P = 0.014; HR, 7.7; 95% CI, 1.5-41.3). These findings strongly suggest that p-TOPK may be a potent biomarker to determine the outcome of patients with PCNSL and to develop novel drugs to treat PCNSL.

Targeting the T-Lak cell originated protein kinase by OTS964 shrinks the size of power-law coded heterogeneous glioma stem cell populations.

Glioblastoma resists chemoradiotherapy, then, recurs to be a fatal space-occupying lesion. The recurrence is caused by re-growing cell populations such as glioma stem cells (GSCs), suggesting that GSC populations should be targeted. This study addressed whether a novel anti-cancer drug, OTS964, an inhibitor for T-LAK cell originated protein kinase (TOPK), is effective in reducing the size of the heterogeneous GSC populations, a power-law coded heterogeneous GSC populations consisting of glioma sphere (GS) clones, by detailing quantitative growth properties. We found that OTS964 killed GS clones while suppressing the growth of surviving GS clones, thus identifying clone-eliminating and growth-disturbing efficacies of OTS964. The efficacies led to a significant size reduction in GS populations in a dose-dependent manner. The surviving GS clones reconstructed GS populations in the following generations; the recovery of GS populations fits a recurrence after the chemotherapy. The recovering GS clones resisted the clone-eliminating effect of OTS964 in sequential exposure during the growth recovery. However, surprisingly, the resistant properties of the recovered-GS clones had been plastically canceled during self-renewal, and then the GS clones had become re-sensitive to OTS964. Thus, OTS964 targets GSCs to eliminate them or suppress their growth, resulting in shrinkage of the power-law coded GSC populations. We propose a therapy focusing on long-term control in recurrence of glioblastoma via reducing the size of the GSC populations by OTS964.

Impact of a novel biomarker, T-LAK cell-originating protein kinase (TOPK) expression on outcome in malignant glioma.

This study aimed to evaluate the biological features of T-lymphokine-activated killer cell-originating protein kinase (TOPK) in vitro and to assess clinical impact of TOPK on the outcome in patients with malignant glioma. TOPK protein level and TOPK mRNA and protein levels in six glioma cell lines were examined using Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. Immunohistochemistry was performed to examine their subcellular localization of TOPK. Using surgical specimens from 57 patients with gliomas, TOPK and Ki-67 expressions were examined by immunohistochemistry. Their co-localization was also examined with double immunofluorescence immunohistochemistry. Impacts of TOPK/Ki-67 expression on the overall survival (OS) and progression-free survival (PFS) in 32 patients with glioblastoma multiforme (GBM) were examined, using Kaplan-Meier and Cox proportion hazard models. Immunohistochemistry revealed that approximately 20-30% of glioma cells were positive for TOPK in vitro. TOPK mRNA was identified in all glioma cell lines on RT-PCR. The value of TOPK/GAPDH was 0.27 ± 0.11. TOPK and Ki-67 expressions were significantly higher in GBM patients than in non-GBM patients. A majority of TOPK-positive cells were also positive for Ki-67 and vice versa. Multivariate analysis revealed that a low TOPK expression (≤ 12.7%) was an independent predictor of longer OS (P = 0.0372), and that gross total removal and a low TOPK expression (≤ 12.7%) were independent predictors of longer PFS (P = 0.0470 and P = 0.0189, respectively). The findings strongly suggest biological and clinical importance of TOPK expression in gliomas, indicating a novel therapeutic potential of TOPK inhibitors to treat malignant gliomas.

Mobilization of Pluripotent Multilineage-Differentiating Stress-Enduring Cells in Ischemic Stroke.

GOAL: This prospective study was aimed to prove the hypothesis that multilineage-differentiating stress-enduring (Muse) cells are mobilized from bone marrow into peripheral blood in patients with ischemic stroke. MATERIALS AND METHODS: This study included 29 patients with ischemic stroke. To quantify the circulating Muse cells, peripheral blood was obtained from all patients on admission and at days 7 and 30. Using fluorescence-activated cell sorting, Muse cells were identified as stage-specific embryonic antigen-3-positive cells. The control values were obtained from 5 healthy volunteers. Separately, immunohistochemistry was performed to evaluate the distribution of Muse cells in the bone marrow of 8 autopsy cases. FINDINGS: The number of Muse cells robustly increased within 24 hours after the onset, compared with the controls, but their baseline number and temporal profile widely varied among patients. No clinical data predicted the baseline number of Muse cells at the onset. Multivariate analysis revealed that smoking and alcohol intake significantly affect the increase in circulating Muse cells. The odds ratio was .0027 (P = .0336) and 1688 (P = .0220) for smoking and alcohol intake, respectively. The percentage of Muse cells in the bone marrow was .20% ± .17%. CONCLUSION: This study shows that pluripotent Muse cells are mobilized from the bone marrow into peripheral blood in the acute stage of ischemic stroke. Smoking and alcohol intake significantly affect their temporal profile. Therapeutic interventions that increase endogenous Muse cells or exogenous administration of Muse cells may improve functional outcome after ischemic stroke.

Involvement of circulating endothelial progenitor cells in carotid plaque growth and vulnerability.

OBJECTIVE The roles of endothelial progenitor cells (EPCs) in the development of carotid plaque are still obscure. This study aimed to clarify this by assessing the histological findings of specimens obtained from carotid endarterectomy. METHODS This study included 34 patients who underwent carotid endarterectomy. MR imaging was performed to semiquantitatively analyze the components of the carotid plaques in all patients. The surgical specimens were subjected to immunohistochemistry. The distributions of the CD34-, CD133-, VEGF-2R-positive cells in the carotid plaques were precisely analyzed, and their number was quantified. Simultaneously, the CD34-positive microvessels were localized. RESULTS The plaque component was judged as lipid-rich plaque in 19 patients, intraplaque hemorrhage (IPH) in 11 patients, and fibrous plaque in 4 patients. The CD34-positive microvessels were densely distributed in the plaque shoulder and interface-to-media regions. The CD34-, CD133-, and VEGF-2R-positive cells were mainly localized around the CD34-positive microvessels. The number of CD34-positive microvessels significantly correlated with the number of CD34-, CD133-, and VEGF-2R-positive cells (R = 0.308, p = 0.009; R = 0.324, p = 0.006; and R = 0.296, p = 0.013, respectively). Vulnerable plaques (lipid-rich and IPH) had significantly higher numbers of the CD34-positive microvessels (p = 0.007) and CD34-, CD133-, and VEGF-2R-positive cells than fibrous plaques (p = 0.031, p = 0.013, and p = 0.002). CONCLUSIONS These findings strongly suggest that neovascularization in the plaque shoulder and interface-to-media regions may play a key role in delivering EPCs from the peripheral blood to the carotid plaque, promoting the growth of carotid plaque. Furthermore, the invaded EPCs, especially the CD133-positive immature EPCs, may be related to plaque vulnerability.

Ischemic stroke in pediatric moyamoya disease associated with immune thrombocytopenia--a case report.

CASE REPORT: A 10-year-old boy developed refractory bleeding and was diagnosed with immune thrombocytopenia (ITP). He was treated with steroids and intravenous immunoglobulin (IVIG). Five months later, however, he developed right homonymous hemianopsia, sensory aphasia, agraphia, and agnosia. MR imaging demonstrated multiple cerebral infarction in the bilateral cerebral hemispheres, and MR angiography revealed severe stenosis of the bilateral internal carotid arteries. He was diagnosed with moyamoya disease and successfully underwent surgical revascularization on both sides under IVIG therapy. However, multiple cerebral infarcts developed in the bilateral cerebral hemispheres 10 days after the second surgical revascularization when platelet counts were within normal limits. Furthermore, chronic subdural hematoma gradually increased in size after each surgery, which required burr hole surgery to resolve increased intracranial pressure, when platelet counts decreased to less than 10 × 10(9)/L. CONCLUSION: This is the first report presenting a case with moyamoya disease coincident with ITP. Critical managements would be essential to reduce perioperative complications, because ITP is known to provoke both hemorrhagic and ischemic events through multiple mechanisms.

Surgical anatomy and preservation of the middle meningeal artery during bypass surgery for moyamoya disease.

BACKGROUND: The middle meningeal artery (MMA) is known to function as one of the important collateral routes in moyamoya disease. However, the anterior branch frequently courses within the lesser wing of the sphenoid bone and can easily be damaged during craniotomy for bypass surgery. This prospective study aimed to study the surgical anatomy of the MMA and to establish the technique to preserve it during bypass surgery for moyamoya disease. METHODS: Twenty-two patients with moyamoya disease underwent STA-MCA anastomosis combined with indirect bypass on 27 sides. The anatomical relationship between the anterior branch of the MMA and lesser wing was classified into three types: the bridge, monorail, and tunnel types. During surgery, the lesser wing was carefully resected with a rongeur or high-speed diamond drill to preserve the anterior branch of the MMA. RESULTS: The anterior branch of the MMA was classified into the bridge type in 5 sides (18.5 %), monorail type in 10 sides (37.0 %), and tunnel type in 12 sides (44.5 %). Patient age was closely related to the anatomical findings (χ (2) test, p = 0.0168). Careful resection of the lesser wing with a rongeur could preserve bridge- and monorail-type MMAs (100 and 71.4 %, respectively). However, drilling out of the lesser wing under a surgical microscope was essential to preserve the tunnel-type MMA. Intraoperative indocyanine green videoangiography was useful to confirm patency during surgery. CONCLUSIONS: It is essential to understand the surgical anatomy of the MMA around the pterion in order to preserve its anterior branch during bypass surgery for moyamoya disease.

[Recurrent hyperperfusion after revascularization surgery for adult moyamoya disease: a case report].

Direct bypass surgery for moyamoya disease is quite useful for rapidly improving cerebral hemodynamics and resolving ischemic attacks but may induce hyperperfusion syndrome. In this report, we present a rare case of recurrent hyperperfusion after surgery for moyamoya disease. A 47-year-old woman developed left homonymous hemianopsia and was admitted to our hospital. Magnetic resonance(MR)imaging/angiography revealed definitive moyamoya disease presenting with acute cerebral infarction in the right temporoparietal lobe. She was treated with anti-hypertensive agents because she had severe hypertension. She successfully underwent right superficial temporal artery-middle cerebral artery double anastomosis and indirect bypass. A cerebral blood flow study just after surgery demonstrated marked hyperperfusion in the operated hemisphere. A repeat study 2 days later revealed that the hyperperfusion was dramatically improved. Subsequently, however, she developed a severe ipsilateral headache and focal seizure regardless of the blood pressure control. She complained of the headache for >2 weeks. Follow-up MRI showed a small subcortical hemorrhage in the right frontal lobe. Single-photon emission computed tomography revealed that the hyperperfusion relapsed 9 days after surgery and then gradually disappeared. This study shows that preoperative severe hypertension may induce uncontrollable hyperperfusion after direct bypass surgery for moyamoya disease.

Cavernous sinus cavernous hemangioma largely extending into the sella turcica and mimicking pituitary adenoma: case report.

A 77-year-old female presented with a rare cavernous sinus cavernous hemangioma with extension to the sella turcica, neuroradiologically mimicking nonfunctioning pituitary macroadenoma. The lesion was partially removed via transsphenoidal surgery, and the histological diagnosis was cavernous hemangioma. After stereotactic radiosurgery using a cyber knife, the lesion decreased in size. Stereotactic radiosurgery may be a good option for cavernous sinus cavernous hemangioma with high risk of surgical bleeding.