Papers - Misawa, Hidemi

ALS/FTD-linked TBK1 deficiency in microglia induces an aged-like microglial signature and drives social recognition deficits in mice.

Lonoel, I., Ribon, M., Lorenc, F., Diebold, A., Philibert, C-E., Robaldo, D., Basdi, M., Perronnet, J., Lameth, J., Berriat, F., Misawa, H., Coutelier, M., Cassel, R., Sarrazin, N., Jost-Mousseau, C., Bohl, D., Millecamps, S., Mallet, M., Brenner, D., Weishaupt, J-H., Boillée, S. and Lobsiger, C-S.

Nat. Commun. 16   7951 2025.08

Research paper (scientific journal), Joint Work, Accepted

CCN1 derived from vascular endothelial cells impairs cognitive function in Alzheimer’s disease model mice.

Hirabayashi, H., Uyeda, A., Manaba, I., Yonezu, Y., Saito, T., Saido, T-C., Misawa, H., Ogasawara, Y., Kinoshita, K. and Muramatsu, R.

J. Pharmacol. Sci. 157   146 - 155 2025.01

Research paper (scientific journal), Joint Work, Accepted

X-ray fluoroscopy-based kinematic analysis of quadrupedal locomotion in slow and fast fatigue-resistant motor neuron-deleted mice.

Ono, A., Inomata, D., Ohgaki, L., Koyama, T., Maeno, A., Misawa, H. Ogihara, N.

Muscle & Nerve 71   257 - 264 2024.12

Research paper (scientific journal), Joint Work, Corresponding author, Accepted

NDRG1 upregulation by ubiquitin proteasome system dysfunction aggravates neurodegeneration.

Hoshino, T., Mukai, A., Yamashita, H., Misawa, H., Urushitani, M., Tashiro, Y., Matsuzawa, S. Takahashi, R.

Mol. Brain 17   77 2024.10

Research paper (scientific journal), Joint Work, Accepted

Type selective ablation of postnatal slow and fast fatigue-resistant motor neurons in mice induces late onset kinetic and postural tremor following fiber-type transition and myopathy.

Misawa, H., Kamishima, K., Koyama, T., Ohgaki, L., Morisaki, Y., Yamanaka, K., Itohara, S., Sawano, S., Mizunoya, W. Ogihara, N.

Exp. Neurol. 376   114772 2024.04

Research paper (scientific journal), Joint Work, Lead author, Corresponding author, Accepted

Suppression of neuroinflammation by coffee component pyrocatechol via inhibition of NF-κB in microglia.

Murata, T., Tago, K., Miyata, K., Moriwaki, Y., Misawa, H., Kobata, K., Nakazawa, Y., Tamura, H. Funakoshi-Tago, M.

Int. J. Mol. Sci. 25   316 2023.12

Research paper (scientific journal), Joint Work, Accepted

Depletion of perivascular macrophages delays ALS disease progression by ameliorating blood-spinal cord barrier impairment in SOD1G93A mice

Adachi K, Miyata K, Chida Y, Hirose M, Morisaki Y, Yamanaka K, Misawa H

Front Cell Neurosci 17   1291673 2023.11

Research paper (scientific journal), Joint Work, Last author, Corresponding author, Accepted

LAG-3 expression in microglia regulated by IFN-γ/STAT1 pathway and metalloproteases

Morisaki Y, Ohshima M, Suzuki H, Misawa H

Front Cell Neurosci 17   1308972 2023.11

Research paper (scientific journal), Joint Work, Last author, Corresponding author, Accepted

GTS-21 enhances regulatory T cell development from T cell receptor-activated human CD4+ T cells exhibiting varied levels of CHRNA7 and CHRFAM7A expression

Mashimo M, Fujii T, Ono S, Moriwaki Y, Misawa H, Azami T, Kasahara T, Kawashima K

Int J Mol Sci 24   12257 2023.07

Research paper (scientific journal), Joint Work, Accepted

Lysophosphatidic acid stimulates pericyte migration via LPA receptor 1

Yonezu Y, Tanabe S, Misawa H, Muramatsu R

Biochem Biophys Res Commun 618   61 - 66 2022.08

Research paper (scientific journal), Joint Work, Accepted

Involvement of neuronal and muscular Trk-fused gene (TFG) defects in the development of neurodegenerative diseases.

Yamamotoya T, Hasei S, Akasaka Y, Ohata Y, Nakatsu Y, Kanna M, Fujishiro M, Sakoda H, Ono H, Kushiyama A, Misawa H, Asano T

Scientific reports 12 ( 1 ) 1966 2022.02

Research paper (scientific journal), Joint Work, Accepted

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Regulation of immune functions by non-neuronal acetylcholine (Ach) via muscarinic and nicotinic ach receptors

Mashimo M, Moriwaki Y, Misawa H, Kawashima K, Fujii T

International Journal of Molecular Sciences (International Journal of Molecular Sciences)  22 ( 13 )  2021.07

Research paper (scientific journal), Joint Work, Accepted,  ISSN  16616596

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Acetylcholine (ACh) is the classical neurotransmitter in the cholinergic nervous system. However, ACh is now known to regulate various immune cell functions. In fact, T cells, B cells, and macrophages all express components of the cholinergic system, including ACh, muscarinic, and nicotinic ACh receptors (mAChRs and nAChRs), choline acetyltransferase, acetylcholinesterase, and choline transporters. In this review, we will discuss the actions of ACh in the immune system. We will first briefly describe the mechanisms by which ACh is stored in and released from immune cells. We will then address Ca2+ signaling pathways activated via mAChRs and nAChRs on T cells and B cells, highlighting the importance of ACh for the function of T cells, B cells, and macrophages, as well as its impact on innate and acquired (cellular and humoral) immunity. Lastly, we will discuss the effects of two peptide ligands, secreted lymphocyte antigen-6/urokinase-type plasminogen activator receptor-related peptide-1 (SLURP-1) and hippocampal cholinergic neurostimulating peptide (HCNP), on cholinergic activity in T cells. Overall, we stress the fact that ACh does not function only as a neurotransmitter; it impacts immunity by exerting diverse effects on immune cells via mAChRs and nAChRs.

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Competitive inhibition of the high-affinity choline transporter by tetrahydropyrimidine anthelmintics

Okuda T, Nomura Y, Konishi A, Misawa H

European Journal of Pharmacology (European Journal of Pharmacology)  898   173986 2021.05

Research paper (scientific journal), Joint Work, Accepted,  ISSN  00142999

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The high-affinity choline transporter CHT1 mediates choline uptake, the rate-limiting and regulatory step in acetylcholine synthesis at cholinergic presynaptic terminals. CHT1-medated choline uptake is specifically inhibited by hemicholinium-3, which is a type of choline analog that acts as a competitive inhibitor. Although the substrate choline and the inhibitor hemicholinium-3 are well-established ligands of CHT1, few potent ligands other than choline analogs have been reported. Here we show that tetrahydropyrimidine anthelmintics, known as nicotinic acetylcholine receptor agonists, act as competitive inhibitors of CHT1. A ligand-dependent trafficking assay in cell lines expressing human CHT1 was designed to search for CHT1 ligands from a collection of biologically active compounds. We found that morantel as well as other tetrahydropyrimidines, pyrantel and oxantel, potently inhibits the high-affinity choline uptake activity of CHT1 in a competitive manner similar to the inhibitor hemicholinium-3. They also inhibit the high-affinity choline transporter from the nematode Caenorhabditis elegans. Finally, tetrahydropyrimidines potently inhibit the high-affinity choline uptake in rat brain synaptosomes at a low micromolar level, resulting in the inhibition of acetylcholine synthesis. The rank order of potency in synaptosomes is as follows: morantel > pyarantel > oxantel (Ki = 1.3, 5.7, and 8.3 μM, respectively). Our results reveal that tetrahydropyrimidine anthelmintics are novel CHT1 ligands that inhibit the high-affinity choline uptake for acetylcholine synthesis in cholinergic neurons.

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Stagnation of glymphatic interstitial fluid flow and delay in waste clearance in the SOD1-G93A mouse model of ALS

Hirose M, Asano M, Watanabe-Matsumoto S, Yamanaka K, Abe Y, Yasui M, Tokuda E, Furukawa Y, Misawa H

Neuroscience Research (Neuroscience Research)  171   74 - 82 2021

Research paper (scientific journal), Joint Work, Accepted,  ISSN  01680102

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© 2020 The Author(s) Overexpression and mislocalization of aquaporin-4 (AQP4) in the SOD1G93A mouse model of amyotrophic lateral sclerosis (ALS) have previously been reported. However, how alterations of AQP4 affect interstitial bulk flow in the brain and spinal cord, the so-called glymphatic system, is unclear. Here, we report an enhanced accumulation of disease-associated SOD1 species including SOD1 oligomers in SOD1G93A;AQP4−/− mice compared with SOD1G93A mice during ALS disease progression, as analyzed by sandwich ELISA. By directly injecting SOD1 oligomers into the spinal cord parenchyma, we observed a significantly larger delay in clearance of biotinylated or fluorescent-labeled SOD1 oligomers in AQP4−/− mice than in wild-type mice. Furthermore, when we injected the fluorescent-labeled tracer protein ovalbumin into the cisterna magna and analyzed the tracer distribution in the cervical spinal cord, approximately 35 % processing ability was found to be reduced in SOD1G93A mice compared to wild-type mice. These results suggest that the glymphatic system is abnormal and that waste clearance is delayed in SOD1G93A mice.

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Endogenous neurotoxin-like protein Ly6H inhibits alpha7 nicotinic acetylcholine receptor currents at the plasma membrane

Moriwaki Y, Kubo N, Watanabe M, Asano S, Shinoda T, Sugino T, Ichikawa D, Tsuji S, Kato F, Misawa H

Scientific Reports 10 ( 1 ) 11996 - 11996 2020.12

Research paper (scientific journal), Joint Work, Accepted

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© 2020, The Author(s). α7 nicotinic acetylcholine receptors (nAChRs) are widely expressed in the central nervous system and regarded as potential therapeutic targets for neurodegenerative conditions, such as Alzheimer’s disease and schizophrenia. Yet, despite the assumed pathophysiological importance of the α7 nAChR, molecular physiological characterization remains poorly advanced because α7 nAChR cannot be properly folded and sorted to the plasma membranes in most mammalian cell lines, thus preventing the analyses in heterologous expression system. Recently, ER-resident membrane protein NACHO was discovered as a strong chaperone for the functional expression of α7 nAChR in non-permissive cells. Ly6H, a brain-enriched GPI-anchored neurotoxin-like protein, was reported as a novel modulator regulating intracellular trafficking of α7 nAChR. In this study, we established cell lines that stably and robustly express surface α7 nAChR by introducing α7 nAChR, Ric-3, and NACHO cDNA into HEK293 cells (Triple α7 nAChR/RIC-3/NACHO cells; TARO cells), and re-evaluated the function of Ly6H. We report here that Ly6H binds with α7 nAChRs on the cell membrane and modulates the channel activity without affecting intracellular trafficking of α7 nAChR.

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Lysophosphatidic acid stimulates pericyte migration via LPA receptor 1.

Yonezu, Y., Tanabe, S., Misawa, H., Muramatsu, R.

Biochem. Biophys. Res. Commun. 618   61 - 66 2020.08

Research paper (scientific journal), Joint Work, Accepted

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Behavioral and electrophysiological evidence for a neuroprotective role of aquaporin-4 in the 5xFAD transgenic mice model

Abe Y, Ikegawa N, Yoshida K, Muramatsu K, Hattori S, Kawai K, Murakami M, Tanaka T, Goda W, Goto M, Yamamoto T, Hashimoto T, Yamada K, Shibata T, Misawa H, Mimura M, Tanaka K.F, Miyakawa T, Iwatsubo T, Hata J.I, Niikura T, Yasui M

Acta Neuropathologica Communications 8 ( 1 ) 67 - 67 2020.05

Research paper (scientific journal), Joint Work, Accepted

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© 2020 The Author(s). Aquaporin-4 (AQP4) has been suggested to be involved in the pathogenesis of neurodegenerative diseases including Alzheimer's disease (AD), which may be due to the modulation of neuroinflammation or the impairment of interstitial fluid bulk flow system in the central nervous system. Here, we show an age-dependent impairment of several behavioral outcomes in 5xFAD AQP4 null mice. Twenty-four-hour video recordings and computational analyses of their movement revealed that the nighttime motion of AQP4-deficient 5xFAD mice was progressively reduced between 20 and 36 weeks of age, with a sharp deterioration occurring between 30 and 32 weeks. This reduction in nighttime motion was accompanied by motor dysfunction and epileptiform neuronal activities, demonstrated by increased abnormal spikes by electroencephalography. In addition, all AQP4-deficient 5xFAD mice exhibited convulsions at least once during the period of the analysis. Interestingly, despite such obvious phenotypes, parenchymal amyloid β (Aβ) deposition, reactive astrocytosis, and activated microgliosis surrounding amyloid plaques were unchanged in the AQP4-deficient 5xFAD mice relative to 5xFAD mice. Taken together, our data indicate that AQP4 deficiency greatly accelerates an age-dependent deterioration of neuronal function in 5xFAD mice associated with epileptiform neuronal activity without significantly altering Aβ deposition or neuroinflammation in this mouse model. We therefore propose that there exists another pathophysiological phase in AD which follows amyloid plaque deposition and neuroinflammation and is sensitive to AQP4 deficiency.

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Minireview: Divergent roles of α7 nicotinic acetylcholine receptors expressed on antigen-presenting cells and CD4⁺ T cells in the regulation of T cell differentiation

Mashimo M, Fujii T, Ono S, Moriwaki Y, Misawa H, Kawashima K

International Immunopharmacology 82   106306 - 106306 2020.05

Research paper (scientific journal), Joint Work, Accepted,  ISSN  15675769

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© 2020 Elsevier B.V. α7 nAChRs expressed on immune cells regulate antigen-specific antibody and proinflammatory cytokine production. Using spleen cells from ovalbumin (OVA)-specific T cell receptor transgenic DO11.10 mice and the α7 nAChR agonist GTS-21, investigation of (1) antigen processing-dependent and (2) -independent, antigen presenting cell (APC)-dependent, naïve CD4+ T cell differentiation, as well as (3) non-specific APC-independent, anti-CD3/CD28 mAbs-induced CD4+ T cell differentiation, revealed the differential roles of α7 nAChRs expressed on T cells and APCs in the regulation of CD4+ T cell differentiation. GTS-21 suppressed OVA-induced antigen processing- and APC-dependent differentiation into regulatory T cells (Tregs) and effector T cells (Th1, Th2 and Th17) without affecting OVA uptake or cell viability. By contrast, GTS-21 upregulated OVA peptide-induced antigen processing-independent T cell differentiation into all lineages. During anti-CD3/CD28 mAbs-induced T cell differentiation in the presence of polarizing cytokines, GTS-21 promoted wild-type T cell differentiation into all lineages, but did not affect α7 nAChR-deficient T cell differentiation. These results demonstrate (1) that α7 nAChRs on APCs downregulate T cell differentiation by inhibiting antigen processing and thereby interfering with antigen presentation; and (2) that α7 nAChRs on T cells upregulate differentiation into Tregs and effector T cells. Thus, the divergent roles of α7 nAChRs on APCs and T cells likely regulate the intensity of immune responses. These findings suggest the possibility of using α7 nAChR agonists to harvest greater numbers of Tregs and Th1 and Th2 cells for adoptive immune therapies for treatment of autoimmune diseases and cancers.

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Oxidative misfolding of Cu/Zn-superoxide dismutase triggered by non-canonical intramolecular disulfide formation

Anzai I., Tokuda E., Handa S., Misawa H., Akiyama S., Furukawa Y.

Free Radical Biology and Medicine (Free Radical Biology and Medicine)  147   187 - 199 2020.02

Research paper (scientific journal), Joint Work, Accepted,  ISSN  08915849

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© 2019 Elsevier Inc. Misfolded Cu/Zn-superoxide dismutase (SOD1) is a pathological species in a subset of amyotrophic lateral sclerosis (ALS). Oxidative stress is known to increase in affected spinal cords of ALS and is thus considered to cause damages on SOD1 leading to the misfolding and aggregation. Despite this, it still remains elusive what triggers misfolding of SOD1 under oxidizing environment. Here, we show that a thiol group of Cys111 in SOD1 is oxidized to a sulfenic acid with hydrogen peroxide and reveal that further dissociation of the bound metal ions from the oxidized SOD1 allows another free Cys residue (Cys6) to nucleophilically attack the sulfenylated Cys111. As a result, an intra-molecular disulfide bond forms between Cys6 and Cys111. Such an abnormal SOD1 with the non-canonical disulfide bond was conformationally extended with significant cytotoxicity as well as high propensity to aggregate. Taken together, we propose a new model of SOD1 misfolding under oxidizing environment, in which formation of the non-canonical intramolecular disulfide bond plays a pivotal role.

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Distinct roles of α7 nAChRs in antigen-presenting cells and CD4⁺ T cells in the regulation of T cell differentiation

Mashimo M, Komori M, Matsui Y, Murase M, Fujii T, Takeshima S, Okuyama H, Ono S, Moriwaki Y, Misawa H, Kawashima K

Frontiers in Immunology (Frontiers in Immunology)  10 ( MAY ) 1102 - 1102 2019

Research paper (scientific journal), Joint Work, Accepted

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Copyright © 2019 Mashimo, Komori, Matsui, Murase, Fujii, Takeshima, Okuyama, Ono, Moriwaki, Misawa and Kawashima. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. It is now apparent that immune cells express a functional cholinergic system and that α7 nicotinic acetylcholine receptors (α7 nAChRs) are involved in regulating T cell differentiation and the synthesis of antigen-specific antibodies and proinflammatory cytokines. Here, we investigated the specific function α7 nAChRs on T cells and antigen presenting cells (APCs) by testing the effect of GTS-21, a selective α7 nAChR agonist, on differentiation of CD4+ T cells from ovalbumin (OVA)-specific TCR transgenic DO11.10 mice activated with OVA or OVA peptide323−339 (OVAp). GTS-21 suppressed OVA-induced antigen processing-dependent development of CD4+ regulatory T cells (Tregs) and effector T cells (Th1, Th2, and Th17). By contrast, GTS-21 up-regulated OVAp-induced antigen processing-independent development of CD4+ Tregs and effector T cells. GTS-21 also suppressed production of IL-2, IFN-γ, IL-4, IL-17, and IL-6 during OVA-induced activation but, with the exception IL-2, enhanced their production during OVAp-induced activation. In addition, during antigen-nonspecific, APC-independent anti-CD3/CD28 antibody-induced CD4+ polyclonal T cell activation in the presence of respective polarizing cytokines, GTS-21 promoted development of all lineages, which indicates that GTS-21 also acts via α7 nAChRs on T cells. These results suggest 1) that α7 nAChRs on APCs suppress CD4+ T cell activation by interfering with antigen presentation through inhibition of antigen processing; 2) that α7 nAChRs on CD4+ T cells up-regulate development of Tregs and effector T cells; and that α7 nAChR agonists and antagonists could be potentially useful agents for immune response modulation and enhancement.

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