14 results on '"Inoue, Haruhisa"'
Search Results
2. Rac-GAP α-Chimerin Regulates Motor-Circuit Formation as a Key Mediator of EphrinB3/EphA4 Forward Signaling
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Iwasato, Takuji, Katoh, Hironori, Nishimaru, Hiroshi, Ishikawa, Yukio, Inoue, Haruhisa, Saito, Yoshikazu M., Ando, Reiko, Iwama, Mizuho, Takahashi, Ryosuke, Negishi, Manabu, and Itohara, Shigeyoshi
- Subjects
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NEURAL circuitry , *PROTEINS , *NEURONS , *TRANSGENES - Abstract
Summary: The ephrin/Eph system plays a central role in neuronal circuit formation; however, its downstream effectors are poorly understood. Here we show that α-chimerin Rac GTPase-activating protein mediates ephrinB3/EphA4 forward signaling. We discovered a spontaneous mouse mutation, miffy (mfy), which results in a rabbit-like hopping gait, impaired corticospinal axon guidance, and abnormal spinal central pattern generators. Using positional cloning, transgene rescue, and gene targeting, we demonstrated that loss of α-chimerin leads to mfy phenotypes similar to those of EphA4 −/− and ephrinB3 −/− mice. α-chimerin interacts with EphA4 and, in response to ephrinB3/EphA4 signaling, inactivates Rac, which is a positive regulator of process outgrowth. Moreover, downregulation of α-chimerin suppresses ephrinB3-induced growth cone collapse in cultured neurons. Our findings indicate that ephrinB3/EphA4 signaling prevents growth cone extension in motor circuit formation via α-chimerin-induced inactivation of Rac. They also highlight the role of a Rho family GTPase-activating protein as a key mediator of ephrin/Eph signaling. [Copyright &y& Elsevier]
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- 2007
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3. One-step induction of photoreceptor-like cells from human iPSCs by delivering transcription factors.
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Otsuka Y, Imamura K, Oishi A, Kondo T, Suga M, Yada Y, Shibukawa R, Okanishi Y, Sagara Y, Tsukita K, Tsujikawa A, and Inoue H
- Abstract
Retinal dystrophies (RDs) lead to irreversible vision impairment with no radical treatment. Although photoreceptor cells (PRCs) differentiated from human induced pluripotent stem cells (iPSCs) are essential for the study of RDs as a scalable source, current differentiation methods for PRCs require multiple steps. To address these issues, we developed a method to generate PRCs from human iPSCs by introducing the transcription factors, CRX and NEUROD1. This approach enabled us to generate induced photoreceptor-like cells (iPRCs) expressing PRC markers. Single-cell RNA sequencing revealed the transcriptome of iPRCs in which the genes associated with phototransduction were expressed. Generated iPRCs exhibited their functional properties in calcium imaging. Furthermore, light-induced damage on iPRCs was inhibited by an antioxidant compound. This simple approach would facilitate the availability of materials for PRC-related research and provide a useful application for disease modeling and drug discovery., Competing Interests: The authors declare no competing interests., (© 2022 The Authors.)
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- 2022
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4. Retraction Notice to: In Vitro Modeling of Blood-Brain Barrier with Human iPSC-Derived Endothelial Cells, Pericytes, Neurons, and Astrocytes via Notch Signaling.
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Yamamizu K, Iwasaki M, Takakubo H, Sakamoto T, Ikuno T, Miyoshi M, Kondo T, Nakao Y, Nakagawa M, Inoue H, and Yamashita JK
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- 2018
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5. iPSC-Based Compound Screening and In Vitro Trials Identify a Synergistic Anti-amyloid β Combination for Alzheimer's Disease.
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Kondo T, Imamura K, Funayama M, Tsukita K, Miyake M, Ohta A, Woltjen K, Nakagawa M, Asada T, Arai T, Kawakatsu S, Izumi Y, Kaji R, Iwata N, and Inoue H
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- Alzheimer Disease drug therapy, Alzheimer Disease immunology, Amyloid beta-Protein Precursor immunology, Drug Evaluation, Preclinical methods, Humans, Alzheimer Disease pathology, Amyloid beta-Peptides immunology, Induced Pluripotent Stem Cells cytology, Neurons pathology
- Abstract
In the process of drug development, in vitro studies do not always adequately predict human-specific drug responsiveness in clinical trials. Here, we applied the advantage of human iPSC-derived neurons, which offer human-specific drug responsiveness, to screen and evaluate therapeutic candidates for Alzheimer's disease (AD). Using AD patient neurons with nearly 100% purity from iPSCs, we established a robust and reproducible assay for amyloid β peptide (Aβ), a pathogenic molecule in AD, and screened a pharmaceutical compound library. We acquired 27 Aβ-lowering screen hits, prioritized hits by chemical structure-based clustering, and selected 6 leading compounds. Next, to maximize the anti-Aβ effect, we selected a synergistic combination of bromocriptine, cromolyn, and topiramate as an anti-Aβ cocktail. Finally, using neurons from familial and sporadic AD patients, we found that the cocktail showed a significant and potent anti-Aβ effect on patient cells. This human iPSC-based platform promises to be useful for AD drug development., (Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.)
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- 2017
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6. Antisense Oligonucleotides Reduce RNA Foci in Spinocerebellar Ataxia 36 Patient iPSCs.
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Matsuzono K, Imamura K, Murakami N, Tsukita K, Yamamoto T, Izumi Y, Kaji R, Ohta Y, Yamashita T, Abe K, and Inoue H
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Spinocerebellar ataxia type 36 is a late-onset, slowly progressive cerebellar syndrome with motor neuron degeneration that is caused by expansions of a hexanucleotide repeat (GGCCTG) in the noncoding region of NOP56 gene, with a histopathological feature of RNA foci formation in postmortem tissues. Here, we report a cellular model using the spinocerebellar ataxia type 36 patient induced pluripotent stem cells (iPSCs). We generated iPSCs from spinocerebellar ataxia type 36 patients and differentiated them into neurons. The number of RNA-foci-positive cells was increased in patient iPSCs and iPSC-derived neurons. Treatment of the 2'-O, 4'-C-ethylene-bridged nucleic acid antisense oligonucleotides (ASOs) targeting NOP56 pre-mRNA reduced RNA-foci-positive cells to ∼50% in patient iPSCs and iPSC-derived neurons. NOP56 mRNA expression levels were lower in patient iPSCs and iPSC-derived neurons than in healthy control neurons. One of the ASOs reduced the number of RNA-foci-positive cells without altering NOP56 mRNA expression levels in patient iPSCs and iPSC-derived neurons. These data show that iPSCs from spinocerebellar ataxia type 36 patients can be useful for evaluating the effects of ASOs toward GGCCTG repeat expansion in spinocerebellar ataxia type 36., (Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.)
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- 2017
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7. In Vitro Modeling of Blood-Brain Barrier with Human iPSC-Derived Endothelial Cells, Pericytes, Neurons, and Astrocytes via Notch Signaling.
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Yamamizu K, Iwasaki M, Takakubo H, Sakamoto T, Ikuno T, Miyoshi M, Kondo T, Nakao Y, Nakagawa M, Inoue H, and Yamashita JK
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- Astrocytes cytology, Biomarkers, Capillary Permeability, Cell Differentiation, Cell Line, Endothelial Cells cytology, Humans, Neurons cytology, Pericytes cytology, Astrocytes metabolism, Blood-Brain Barrier cytology, Blood-Brain Barrier metabolism, Endothelial Cells metabolism, Induced Pluripotent Stem Cells cytology, Neurons metabolism, Pericytes metabolism, Receptors, Notch metabolism, Signal Transduction
- Abstract
The blood-brain barrier (BBB) is composed of four cell populations, brain endothelial cells (BECs), pericytes, neurons, and astrocytes. Its role is to precisely regulate the microenvironment of the brain through selective substance crossing. Here we generated an in vitro model of the BBB by differentiating human induced pluripotent stem cells (hiPSCs) into all four populations. When the four hiPSC-derived populations were co-cultured, endothelial cells (ECs) were endowed with features consistent with BECs, including a high expression of nutrient transporters (CAT3, MFSD2A) and efflux transporters (ABCA1, BCRP, PGP, MRP5), and strong barrier function based on tight junctions. Neuron-derived Dll1, which activates Notch signaling in ECs, was essential for the BEC specification. We performed in vitro BBB permeability tests and assessed ten clinical drugs by nanoLC-MS/MS, finding a good correlation with the BBB permeability reported in previous cases. This technology should be useful for research on human BBB physiology, pathology, and drug development., (Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.)
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- 2017
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8. Vulnerability of Purkinje Cells Generated from Spinocerebellar Ataxia Type 6 Patient-Derived iPSCs.
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Ishida Y, Kawakami H, Kitajima H, Nishiyama A, Sasai Y, Inoue H, and Muguruma K
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- 2017
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9. Simple Derivation of Spinal Motor Neurons from ESCs/iPSCs Using Sendai Virus Vectors.
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Goto K, Imamura K, Komatsu K, Mitani K, Aiba K, Nakatsuji N, Inoue M, Kawata A, Yamashita H, Takahashi R, and Inoue H
- Abstract
Amyotrophic lateral sclerosis (ALS) is a progressive and fatal degenerative disorder of motor neurons (MNs). Embryonic stem cells (ESCs)/induced pluripotent stem cells (iPSCs) now help us to understand the pathomechanisms of ALS via disease modeling. Various methods to differentiate ESCs/iPSCs into MNs by the addition of signaling molecules have been reported. However, classical methods require multiple steps, and newer simple methods using the transduction of transcription factors run the risk of genomic integration of the vector genes. Heterogeneity of the expression levels of the transcription factors also remains an issue. Here we describe a novel approach for differentiating human and mouse ESCs/iPSCs into MNs using a single Sendai virus vector encoding three transcription factors, LIM/homeobox protein 3, neurogenin 2, and islet-1, which are integration free. This single-vector method, generating HB9-positive cells on day 2 from human iPSCs, increases the ratio of MNs to neurons compared to the use of three separate Sendai virus vectors. In addition, the MNs derived via this method from iPSCs of ALS patients and model mice display disease phenotypes. This simple approach significantly reduces the efforts required to generate MNs, and it provides a useful tool for disease modeling.
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- 2017
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10. Vulnerability of Purkinje Cells Generated from Spinocerebellar Ataxia Type 6 Patient-Derived iPSCs.
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Ishida Y, Kawakami H, Kitajima H, Nishiyama A, Sasai Y, Inoue H, and Muguruma K
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- Calcium Channels, N-Type chemistry, Calcium Channels, N-Type metabolism, Cell Differentiation drug effects, Humans, Induced Pluripotent Stem Cells drug effects, Induced Pluripotent Stem Cells metabolism, Protein Domains, Purkinje Cells drug effects, Purkinje Cells metabolism, Riluzole pharmacology, Thyrotropin pharmacology, Up-Regulation drug effects, Induced Pluripotent Stem Cells pathology, Purkinje Cells pathology, Spinocerebellar Ataxias pathology
- Abstract
Spinocerebellar ataxia type 6 (SCA6) is a dominantly inherited neurodegenerative disease characterized by loss of Purkinje cells in the cerebellum. SCA6 is caused by CAG trinucleotide repeat expansion in CACNA1A, which encodes Cav2.1, α1A subunit of P/Q-type calcium channel. However, the pathogenic mechanism and effective therapeutic treatments are still unknown. Here, we have succeeded in generating differentiated Purkinje cells that carry patient genes by combining disease-specific iPSCs and self-organizing culture technologies. Patient-derived Purkinje cells exhibit increased levels of full-length Cav2.1 protein but decreased levels of its C-terminal fragment and downregulation of the transcriptional targets TAF1 and BTG1. We further demonstrate that SCA6 Purkinje cells exhibit thyroid hormone depletion-dependent degeneration, which can be suppressed by two compounds, thyroid releasing hormone and Riluzole. Thus, we have constructed an in vitro disease model recapitulating both ontogenesis and pathogenesis. This model may be useful for pathogenic investigation and drug screening., (Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.)
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- 2016
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11. Modeling the early phenotype at the neuromuscular junction of spinal muscular atrophy using patient-derived iPSCs.
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Yoshida M, Kitaoka S, Egawa N, Yamane M, Ikeda R, Tsukita K, Amano N, Watanabe A, Morimoto M, Takahashi J, Hosoi H, Nakahata T, Inoue H, and Saito MK
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- Animals, Cell Culture Techniques, Cell Differentiation, Cell Line, Coculture Techniques, Humans, Induced Pluripotent Stem Cells cytology, Induced Pluripotent Stem Cells drug effects, Mice, Morpholinos pharmacology, Motor Neurons cytology, Motor Neurons metabolism, Muscular Atrophy, Spinal genetics, Neuromuscular Junction drug effects, Neuromuscular Junction pathology, Spinal Muscular Atrophies of Childhood genetics, Spinal Muscular Atrophies of Childhood metabolism, Valproic Acid pharmacology, Induced Pluripotent Stem Cells metabolism, Muscular Atrophy, Spinal metabolism, Neuromuscular Junction metabolism, Phenotype
- Abstract
Spinal muscular atrophy (SMA) is a neuromuscular disorder caused by mutations of the survival of motor neuron 1 (SMN1) gene. In the pathogenesis of SMA, pathological changes of the neuromuscular junction (NMJ) precede the motor neuronal loss. Therefore, it is critical to evaluate the NMJ formed by SMA patients' motor neurons (MNs), and to identify drugs that can restore the normal condition. We generated NMJ-like structures using MNs derived from SMA patient-specific induced pluripotent stem cells (iPSCs), and found that the clustering of the acetylcholine receptor (AChR) is significantly impaired. Valproic acid and antisense oligonucleotide treatment ameliorated the AChR clustering defects, leading to an increase in the level of full-length SMN transcripts. Thus, the current in vitro model of AChR clustering using SMA patient-derived iPSCs is useful to dissect the pathophysiological mechanisms underlying the development of SMA, and to evaluate the efficacy of new therapeutic approaches., (Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2015
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12. Focal transplantation of human iPSC-derived glial-rich neural progenitors improves lifespan of ALS mice.
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Kondo T, Funayama M, Tsukita K, Hotta A, Yasuda A, Nori S, Kaneko S, Nakamura M, Takahashi R, Okano H, Yamanaka S, and Inoue H
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- Amyotrophic Lateral Sclerosis mortality, Amyotrophic Lateral Sclerosis pathology, Animals, Astrocytes cytology, Astrocytes metabolism, Cell Differentiation, Disease Models, Animal, Humans, Kaplan-Meier Estimate, Mice, Mice, Transgenic, Nerve Growth Factors metabolism, Neural Stem Cells cytology, Neural Stem Cells metabolism, Proto-Oncogene Proteins c-akt metabolism, Signal Transduction, Spinal Cord pathology, Superoxide Dismutase genetics, Superoxide Dismutase metabolism, Transplantation, Heterologous, Up-Regulation, Amyotrophic Lateral Sclerosis therapy, Induced Pluripotent Stem Cells cytology, Neural Stem Cells transplantation
- Abstract
Transplantation of glial-rich neural progenitors has been demonstrated to attenuate motor neuron degeneration and disease progression in rodent models of mutant superoxide dismutase 1 (SOD1)-mediated amyotrophic lateral sclerosis (ALS). However, translation of these results into a clinical setting requires a renewable human cell source. Here, we derived glial-rich neural progenitors from human iPSCs and transplanted them into the lumbar spinal cord of ALS mouse models. The transplanted cells differentiated into astrocytes, and the treated mouse group showed prolonged lifespan. Our data suggest a potential therapeutic mechanism via activation of AKT signal. The results demonstrated the efficacy of cell therapy for ALS by the use of human iPSCs as cell source., (Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2014
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13. A chemical probe that labels human pluripotent stem cells.
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Hirata N, Nakagawa M, Fujibayashi Y, Yamauchi K, Murata A, Minami I, Tomioka M, Kondo T, Kuo TF, Endo H, Inoue H, Sato SI, Ando S, Kawazoe Y, Aiba K, Nagata K, Kawase E, Chang YT, Suemori H, Eto K, Nakauchi H, Yamanaka S, Nakatsuji N, Ueda K, and Uesugi M
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- Animals, HEK293 Cells, Humans, Mice, Microscopy, Fluorescence methods, Fluorescent Dyes chemistry, Induced Pluripotent Stem Cells chemistry, Induced Pluripotent Stem Cells cytology, Molecular Probes chemistry, Pluripotent Stem Cells chemistry, Pluripotent Stem Cells cytology
- Abstract
A small-molecule fluorescent probe specific for human pluripotent stem cells would serve as a useful tool for basic cell biology research and stem cell therapy. Screening of fluorescent chemical libraries with human induced pluripotent stem cells (iPSCs) and subsequent evaluation of hit molecules identified a fluorescent compound (Kyoto probe 1 [KP-1]) that selectively labels human pluripotent stem cells. Our analyses indicated that the selectivity results primarily from a distinct expression pattern of ABC transporters in human pluripotent stem cells and from the transporter selectivity of KP-1. Expression of ABCB1 (MDR1) and ABCG2 (BCRP), both of which cause the efflux of KP-1, is repressed in human pluripotent stem cells. Although KP-1, like other pluripotent markers, is not absolutely specific for pluripotent stem cells, the identified chemical probe may be used in conjunction with other reagents., (Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2014
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14. Modeling Alzheimer's disease with iPSCs reveals stress phenotypes associated with intracellular Aβ and differential drug responsiveness.
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Kondo T, Asai M, Tsukita K, Kutoku Y, Ohsawa Y, Sunada Y, Imamura K, Egawa N, Yahata N, Okita K, Takahashi K, Asaka I, Aoi T, Watanabe A, Watanabe K, Kadoya C, Nakano R, Watanabe D, Maruyama K, Hori O, Hibino S, Choshi T, Nakahata T, Hioki H, Kaneko T, Naitoh M, Yoshikawa K, Yamawaki S, Suzuki S, Hata R, Ueno S, Seki T, Kobayashi K, Toda T, Murakami K, Irie K, Klein WL, Mori H, Asada T, Takahashi R, Iwata N, Yamanaka S, and Inoue H
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- Amyloid beta-Peptides chemistry, Cell Differentiation, Cerebral Cortex pathology, Humans, Induced Pluripotent Stem Cells drug effects, Intracellular Space drug effects, Mutant Proteins, Neurons metabolism, Neurons pathology, Phenotype, Protein Structure, Quaternary, Alzheimer Disease pathology, Amyloid beta-Peptides metabolism, Docosahexaenoic Acids pharmacology, Induced Pluripotent Stem Cells metabolism, Intracellular Space metabolism, Models, Biological, Oxidative Stress drug effects
- Abstract
Oligomeric forms of amyloid-β peptide (Aβ) are thought to play a pivotal role in the pathogenesis of Alzheimer's disease (AD), but the mechanism involved is still unclear. Here, we generated induced pluripotent stem cells (iPSCs) from familial and sporadic AD patients and differentiated them into neural cells. Aβ oligomers accumulated in iPSC-derived neurons and astrocytes in cells from patients with a familial amyloid precursor protein (APP)-E693Δ mutation and sporadic AD, leading to endoplasmic reticulum (ER) and oxidative stress. The accumulated Aβ oligomers were not proteolytically resistant, and docosahexaenoic acid (DHA) treatment alleviated the stress responses in the AD neural cells. Differential manifestation of ER stress and DHA responsiveness may help explain variable clinical results obtained with the use of DHA treatment and suggests that DHA may in fact be effective for a subset of patients. It also illustrates how patient-specific iPSCs can be useful for analyzing AD pathogenesis and evaluating drugs., (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Published
- 2013
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