Hidemoto Saiki

Dopaminergic neurons generated from monkey embryonic stem cells function in a Parkinson primate model

Parkinson disease (PD) is a neurodegenerative disorder characterized by loss of midbrain dopaminergic (DA) neurons. ES cells are currently the most promising donor cell source for cell-replacement therapy in PD. We previously described a strong neuralizing activity present on the surface of stromal cells, named stromal cell-derived inducing activity (SDIA). In this study, we generated neurospheres composed of neural progenitors from monkey ES cells, which are capable of producing large numbers of DA neurons. We demonstrated that FGF20, preferentially expressed in the substantia nigra, acts synergistically with FGF2 to increase the number of DA neurons in ES cell-derived neurospheres. We also analyzed the effect of transplantation of DA neurons generated from monkey ES cells into 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated (MPTP-treated) monkeys, a primate model for PD. Behavioral studies and functional imaging revealed that the transplanted cells functioned as DA neurons and attenuated MPTP-induced neurological symptoms.

Altered plasticity of the human motor cortex in Parkinson's disease

Interventional paired associative stimulation (IPAS) to the contralateral peripheral nerve and cerebral cortex can enhance the primary motor cortex (M1) excitability with two synchronously arriving inputs. This study investigated whether dopamine contributed to the associative long‐term potentiation–like effect in the M1 in Parkinsons disease (PD) patients. Eighteen right‐handed PD patients and 11 right‐handed age‐matched healthy volunteers were studied. All patients were studied after 12 hours off medication with levodopa replacement (PD‐off). Ten patients were also evaluated after medication (PD‐on). The IPAS comprised a single electric stimulus to the right median nerve at the wrist and subsequent transcranial magnetic stimulation of the left M1 with an interstimulus interval of 25 milliseconds (240 paired stimuli every 5 seconds for 20 minutes). The motor‐evoked potential amplitude in the right abductor pollicis brevis muscle was increased by IPAS in healthy volunteers, but not in PD patients. IPAS did not affect the motor‐evoked potential amplitude in the left abductor pollicis brevis. The ratio of the motor‐evoked potential amplitude before and after IPAS in PD‐off patients increased after dopamine replacement. Thus, dopamine might modulate cortical plasticity in the human M1, which could be related to higher order motor control, including motor learning. Ann Neurol 2006

Prolonged maturation culture favors a reduction in the tumorigenicity and the dopaminergic function of human ESC-derived neural cells in a primate model of Parkinson's disease

For the safe clinical application of embryonic stem cells (ESCs) for neurological diseases, it is critical to evaluate the tumorigenicity and function of human ESC (hESC)‐derived neural cells in primates. We have herein, for the first time, compared the growth and function of hESC‐derived cells with different stages of neural differentiation implanted in the brains of primate models of Parkinsons disease. We herein show that residual undifferentiated cells expressing ESC markers present in the cell preparation can induce tumor formation in the monkey brain. In contrast, a cell preparation matured by 42‐day culture with brain‐derived neurotrophic factor/glial cell line‐derived neurotrophic factor (BDNF/GDNF) treatment did not form tumors and survived as primarily dopaminergic (DA) neurons. In addition, the monkeys with such grafts showed behavioral improvement for at least 12 months. These results support the idea that hESCs, if appropriately matured, can serve as a source for DA neurons without forming any tumors in a primate brain. STEM CELLS 2012;30:935–945

Survival of human induced pluripotent stem cell-derived midbrain dopaminergic neurons in the brain of a primate model of Parkinson's disease.

Before induced pluripotent stem cells (iPSCs) can be used to treat neurologic diseases, human iPSC-derived neural cells must be analyzed in the primate brain. In fact, although mouse and human iPSCs have been used to generate dopaminergic (DA) neurons that are beneficial in rat models of Parkinsons disease (PD), human iPSC-derived neural progenitor cells (NPCs) have not been examined in primate brains. Here, we generated NPCs at different stages of predifferentiation using a feeder-free culture method, and grafted them into the brains of a monkey PD model and NOD-SCID mice. Magnetic resonance imaging (MRI), positron emission tomography (PET), immunocytochemistry, and behavioral analyses revealed that NPCs pretreated with Sonic hedgehog and fibroblast growth factor-8 followed by glial cell-derived neurotrophic factor, brain-derived neurotrophic factor, ascorbic acid, and dibutyryl cyclic AMP resulted in smaller grafts than those without these treatments, and survived as DA neurons in a monkey brain as long as six months. Thus, for the first time, we describe a feeder-free neural differentiation method from human iPSCs and an evaluation system that can be used to assess monkey PD models.

Capillary beds are decreased in Alzheimer's disease, but not in Binswanger's disease

Morphological abnormalities of the cortical microvessels have been reported in Alzheimers disease (AD), but not in Binswangers disease (BD), a form of vascular dementia. Therefore, we compared the capillary beds in AD and BD brains, using a modified Gallyas silver impregnation method and immunohistochemistry for beta amyloid. Eight autopsied brains with AD and seven with BD were compared with six control brains. The cortical microvessels in AD were frequently narrowed, and torn off, especially in close proximity to the senile plaques. The capillary densities in AD were significantly decreased as compared with the control brains. In contrast, there were no significant changes in the capillary densities and their morphologies in BD brains. Immunohistochemistry for beta amyloid revealed numerous deposits in the vascular wall and perivascular neuropil exclusively in AD brains. Cortical microvascular changes in AD and their absence in BD may indicate a role of beta amyloid for the microvessel pathology in AD.

Objective and quantitative evaluation of motor function in a monkey model of Parkinson's disease

Monkeys treated with 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine (MPTP) are currently the best animal model for Parkinsons disease (PD) and have been widely used for physiological and pharmacological investigations. However, objective and quantitative assessments have not been established for grading their motor behaviors. In order to develop a method for an unbiased evaluation, we performed a video-based assessment, used qualitative rating scales, and carried out an in vivo investigation of dopamine (DA) transporter binding in systemically MPTP-treated monkeys. The video-based analysis of spontaneous movement clearly demonstrated a significant correlation with the qualitative rating score. The assessment of DA transporter (DAT) function by [(11)C]-CFT-PET showed that, when compared with normal animals, the MPTP-treated animals exhibited decreased CFT binding in the bilateral striatum, particularly in the dorsal part in the putamen and caudate. Among the MPTP-treated monkeys, an unbiased PET analysis revealed a significant correlation between CFT binding in the midbrain and qualitative rating scores or the amount of spontaneous movements. These results indicate that a video-based analysis can be a reliable tool for an objective and quantitative evaluation of motor dysfunction of MPTP-treated monkeys, and furthermore, that DAT function in the midbrain may also be important for the evaluation.

A novel composite targeting method using high-field magnetic resonance imaging for subthalamic nucleus deep brain stimulation.

OBJECT Accurate localization of the subthalamic nucleus (STN) is important for proper placement of the electrodes in deep brain stimulation (DBS) surgery for patients with advanced Parkinson disease. The authors evaluated the accuracy of our modified composite targeting method and the value of using high-field MR imaging for targeting the STN. METHODS Thirteen patients with advanced Parkinson disease underwent bilateral STN DBS based on 3-T MR imaging, and 13 patients underwent surgery based on 1.5-T MR imaging. By sequentially referring to the postmammillary commissure, the red nucleus, the mammillothalamic tract, and the STN, the modified composite targeting method determined the stereotactic coordinates for targeting the STN. The accuracy and efficacy of the composite targeting method and 3-T MR imaging were evaluated by using the intraoperative microelectrode recording, the postoperative imaging study, and the postoperative clinical improvement. RESULTS The landmark structures for targeting the STN were visualized clearly with 3-T MR imaging. The mean (+/- SD) path length through the STN of the central track was 4.9 +/- 1.1 mm in the 3-T group and 3.1 +/- 2.0 mm in the 1.5-T group (p < 0.001). Twenty-one (81%) of 26 electrodes were placed in the central track in the 3-T group, whereas 8 (31%) of 26 electrodes were placed in the central track in the 1.5-T group (p = 0.006). The rest of the electrodes were placed in the noncentral optimum track for alleviating parkinsonian motor symptoms. The mean Unified Parkinsons Disease Rating Scale motor part score during off period was reduced by 53% in the 3-T group and by 41% in the 1.5-T group (p = 0.14). The mean reductions of levodopa equivalent daily doses were 48.6% in the 3-T group and 43.7% in the 1.5-T group (p = 0.61). CONCLUSIONS The use of the modified composite targeting method referring to the multiple landmarks with 3-T MR imaging offers reliable and clinically effective target for STN DBS surgery.

Subthalamic nucleus deep brain stimulation restores normal rapid eye movement sleep in Parkinson's disease.

In Parkinsons disease, sleep disturbance is a common occurrence.

Transplantation of embryonic stem cell-derived dopaminergic neurons in MPTP-treated monkeys.

One of the target diseases of cell-replacement therapy is Parkinsons disease. Clinical experiences with fetal dopaminergic cell graft have shown that the therapy is effective, but limited and accompanied by side effects, such as dyskinesia. So, the therapy needs to be further improved and sophisticated. Embryonic stem (ES) cells are expected to be another donor cell for the treatment, because of its proliferative and differentiation capacities. For clinical application, experiments using non-human primates are important, because size, anatomy, and biological characteristics of the brain are different between rodents and primates. Here, we would like to discuss induction of dopaminergic neurons from monkey ES cells and cell transplantation into the brain of monkey Parkinsons disease model.

Neuroleptic malignant syndrome with prolonged catatonia in a dopa-responsive dystonia patient

Abstract—The authors describe a patient with dopa-responsive dystonia who developed neuroleptic malignant syndrome with prolonged catatonia following treatment with neuroleptic agents. Use of these agents probably expanded the patient’s neuronal dysfunction beyond the nigrostriatal system to involve multiple dopaminergic systems. Electroconvulsive treatment alleviated the prolonged catatonia.