Research Areas 【 display / non-display 】

Research Areas 【 display / non-display 】

• Life sciences   Neuroscience - general  

Affiliation 【 display / non-display 】

Affiliation 【 display / non-display 】

• Yale大学   Associate Research Scientist  

Papers 【 display / non-display 】

Papers 【 display / non-display 】

• Human mesenchymal stem-derived extracellular vesicles improve body growth and motor function following severe spinal cord injury in rat.

  Masahito Nakazaki, Karen L Lankford, Hideaki Yamamoto, Yoshiyuki Mae, Jeffery D Kocsis

  Clinical and translational medicine   13 ( 6 ) e1284  2023.06  [International journal]

  　View Summary

  BACKGROUND: Spinal cord injury (SCI) in young adults leads to severe sensorimotor disabilities as well as slowing of growth. Systemic pro-inflammatory cytokines are associated with growth failure and muscle wasting. Here we investigated whether intravenous (IV) delivery of small extracellular vesicles (sEVs) derived from human mesenchymal stem/stromal cells (MSC) has therapeutic effects on body growth and motor recovery and can modulate inflammatory cytokines following severe SCI in young adult rats. METHODS: Contusional SCI rats were randomized into three different treatment groups (human and rat MSC-sEVs and a PBS group) on day 7 post-SCI. Functional motor recovery and body growth were assessed weekly until day 70 post-SCI. Trafficking of sEVs after IV infusions in vivo, the uptake of sEVs in vitro, macrophage phenotype at the lesion and cytokine levels at the lesion, liver and systemic circulation were also evaluated. RESULTS: An IV delivery of both human and rat MSC-sEVs improved functional motor recovery after SCI and restored normal body growth in young adult SCI rats, indicating a broad therapeutic benefit of MSC-sEVs and a lack of species specificity for these effects. Human MSC-sEVs were selectively taken up by M2 macrophages in vivo and in vitro, consistent with our previous observations of rat MSC-sEV uptake. Furthermore, the infusion of human or rat MSC-sEVs resulted in an increase in the proportion of M2 macrophages and a decrease in the production of the pro-inflammatory cytokines tumour necrosis factor-alpha (TNF-α) and interleukin (IL)-6 at the injury site, as well as a reduction in systemic serum levels of TNF-α and IL-6 and an increase in growth hormone receptors and IGF-1 levels in the liver. CONCLUSIONS: Both human and rat MSC-sEVs promote the recovery of body growth and motor function after SCI in young adult rats possibly via the cytokine modulation of growth-related hormonal pathways. Thus, MSC-sEVs affect both metabolic and neurological deficits in SCI.

  DOI PubMed

• Small extracellular vesicles released by infused mesenchymal stromal cells target M2 macrophages and promote TGF-β upregulation, microvascular stabilization and functional recovery in a rodent model of severe spinal cord injury.

  Masahito Nakazaki, Tomonori Morita, Karen L Lankford, Philip W Askenase, Jeffery D Kocsis

  Journal of extracellular vesicles   10 ( 11 ) e12137  2021.09  [International journal]

  　View Summary

  Intravenous (IV) infusion of bone marrow-derived mesenchymal stem/stromal cells (MSCs) stabilizes the blood-spinal cord barrier (BSCB) and improves functional recovery in experimental models of spinal cord injury (SCI). Although IV delivered MSCs do not traffic to the injury site, IV delivered small extracellular vesicles (sEVs) derived from MSCs (MSC-sEVs) do and are taken up by a subset of M2 macrophages. To test whether sEVs released by MSCs are responsible for the therapeutic effects of MSCs, we tracked sEVs produced by IV delivered DiR-labelled MSCs (DiR-MSCs) after transplantation into SCI rats. We found that sEVs were released by MSCs in vivo, trafficked to the injury site, associated specifically with M2 macrophages and co-localized with exosome markers. Furthermore, while a single MSC injection was sufficient to improve locomotor recovery, fractionated dosing of MSC-sEVs over 3 days (F-sEVs) was required to achieve similar therapeutic effects. Infusion of F-sEVs mimicked the effects of single dose MSC infusion on multiple parameters including: increased expression of M2 macrophage markers, upregulation of transforming growth factor-beta (TGF-β), TGF-β receptors and tight junction proteins, and reduction in BSCB permeability. These data suggest that release of sEVs by MSCs over time induces a cascade of cellular responses leading to improved functional recovery.

  DOI PubMed

• Intravenous infusion of auto serum-expanded autologous mesenchymal stem cells in spinal cord injury patients: 13 case series.

  Osamu Honmou, Toshihiko Yamashita, Tomonori Morita, Tsutomu Oshigiri, Ryosuke Hirota, Satoshi Iyama, Junji Kato, Yuichi Sasaki, Sumio Ishiai, Yoichi M Ito, Ai Namioka, Takahiro Namioka, Masahito Nakazaki, Yuko Kataoka-Sasaki, Rie Onodera, Shinichi Oka, Masanori Sasaki, Stephen G Waxman, Jeffery D Kocsis

  Clinical neurology and neurosurgery   203   106565 - 106565  2021.04  [International journal]

  　View Summary

  BACKGROUND: Although spinal cord injury (SCI) is a major cause of disability, current therapeutic options remain limited. Recent progress in cellular therapy with mesenchymal stem cells (MSCs) has provided improved function in animal models of SCI. We investigated the safety and feasibility of intravenous infusion of MSCs for SCI patients and assessed functional status after MSC infusion. METHODS: In this phase 2 study of intravenous infusion of autologous MSCs cultured in auto-serum, a single infusion of MSCs under Good Manufacturing Practice (GMP) production was delivered in 13 SCI patients. In addition to assessing feasibility and safety, neurological function was assessed using the American Spinal Injury Association Impairment Scale (ASIA), International Standards for Neurological and Functional Classification of Spinal Cord (ISCSCI-92). Ability of daily living was assessed using Spinal Cord Independence Measure (SCIM-III). The study protocol was based on advice provided by the Pharmaceuticals and Medical Devices Agency in Japan. The trial was registered with the Japan Medical Association (JMA-IIA00154). RESULTS: No serious adverse events were associated with MSC injection. There was neurologic improvement based on ASIA grade in 12 of the 13 patients at six months post-MSC infusion. Five of six patients classified as ASIA A prior to MSC infusion improved to ASIA B (3/6) or ASIA C (2/6), two ASIA B patients improved to ASIA C (1/2) or ASIA D (1/2), five ASIA C patients improved and reached a functional status of ASIA D (5/5). Notably, improvement from ASIA C to ASIA D was observed one day following MSC infusion for all five patients. Assessment of both ISCSCI-92, SCIM-III also demonstrated functional improvements at six months after MSC infusion, compared to the scores prior to MSC infusion in all patients. CONCLUSION: While we emphasize that this study was unblinded, and does not exclude placebo effects or a contribution of endogenous recovery or observer bias, our observations provide evidence supporting the feasibility, safety and functional improvements of infused MSCs into patients with SCI.

  DOI PubMed

• Intravenous infusion of mesenchymal stem cells inhibits intracranial hemorrhage after recombinant tissue plasminogen activator therapy for transient middle cerebral artery occlusion in rats

  Masahito Nakazaki, Masanori Sasaki, Yuko Kataoka-Sasaki, Shinichi Oka, Takahiro Namioka, Ai Namioka, Rie Onodera, Junpei Suzuki, Yuichi Sasaki, Hiroshi Nagahama, Takeshi Mikami, Masahiko Wanibuchi, Jeffery D. Kocsis, Osamu Honmou

  Journal of Neurosurgery ( Journal of Neurosurgery Publishing Group ({JNSPG}) )  127 ( 4 ) 1 - 10  2017.01  [Refereed]

  　View Summary

  OBJECTIVE Reperfusion therapy with intravenous recombinant tissue plasminogen activator (rtPA) is the standard of care for acute ischemic stroke. However, hemorrhagic complications can result. Intravenous infusion of mesenchymal stem cells (MSCs) reduces stroke volume and improves behavioral function in experimental stroke models. One suggested therapeutic mechanism is inhibition of vascular endothelial dysfunction. The objective of this study was to determine whether MSCs suppress hemorrhagic events after rtPA therapy in the acute phase of transient middle cerebral artery occlusion (tMCAO) in rats. METHODS After induction of tMCAO, 4 groups were studied: 1) normal saline [NSI+vehicle, 2) rtPA+vehicle, 3) NS+MSCs, and 4) rtPA+MSCs. The incidence rate of intracerebral hemorrhage, both hemorrhagic and ischemic volume, and behavioral performance were examined. Matrix metalloproteinase-9 (MMP-9) levels in the brain were assessed with zymography. Quantitative analysis of regional cerebral blood flow (rCBF) was performed to assess hemodynamic change in the ischemic lesion. RESULTS The MSC-treated groups (Groups 3 and 4) experienced a greater reduction in the incidence rate of intracerebral hemorrhage and hemorrhagic volume 1 day after tMCAO even if rtPA was received. The application of rtPA enhanced activation of MMP-9, but MSCs inhibited MMP-9 activation. Behavioral testing indicated that both MSC-infused groups had greater improvement than non-MSC groups had, but rtPA+MSCs provided greater improvement than MSCs alone. The rCBF ratio of rtPA groups (Groups 2 and 4) was similar at 2 hours after reperfusion of tMCAO, but both were greater than that in non-rtPA groups. CONCLUSIONS Infused MSCs may inhibit endothelial dysfunction to suppress hemorrhagic events and facilitate functional outcome. Combined therapy of infused MSCs after rtPA therapy facilitated early behavioral recovery.

  DOI

• Pharmacological Difference Between Platelet Aggregations in Cardioembolic Stroke Patients with Direct Oral Anticoagulants: A Pilot Study

  Masahito Nakazaki, Shinichi Oka, Hirotoshi Magota, Ryo Kiyose, Rie Onodera, Ryo Ukai, Yuko Kataoka-Sasaki, Masanori Sasaki, Osamu Honmou

  Journal of Stroke and Cerebrovascular Diseases ( Elsevier BV )  31 ( 7 ) 106520 - 106520  2022.07

  DOI

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Misc 【 display / non-display 】

Misc 【 display / non-display 】

• 脳梗塞に対する血栓回収療法後の骨髄間葉系幹細胞による細胞治療

  中崎公仁, 佐々木祐典, 佐々木祐典, 佐々木優子, 岡真一, 浪岡隆洋, 浪岡愛, KOCSIS Jeffery, 本望修, 本望修

  脳血管内治療(Web) ( (NPO)日本脳神経血管内治療学会 )  1 ( Supplement ) S221(J‐STAGE) - S221  2016.11

  J-GLOBAL

• Regenerative therapy using autologous mesenchymal stem cell for cerebral ischemia

  中﨑 公仁, 岡 真一, 浪岡 愛

  再生医療 : 日本再生医療学会雑誌 ( メディカルレビュー社 )  13 ( 3 ) 304 - 308  2014.08

  CiNii

• Preservation of interhemispheric cortical connections through corpus callosum following intravenous infusion of mesenchymal stem cells in a rat model of cerebral infarction.

  Nagahama H, Nakazaki M, Sasaki M, Kataoka-Sasaki Y, Namioka T, Namioka A, Oka S, Onodera R, Suzuki J, Sasaki Y, Kocsis JD, Honmou O

  Brain research   1695   37 - 44  2018.05  [Refereed]  [International journal]

  　View Summary

  © 2018 Elsevier B.V. Systemic administration of mesenchymal stem cells (MSCs) following cerebral infarction exerts functional improvements. Previous research has suggested potential therapeutic mechanisms that promote neuroprotection and synaptogenesis. These include secretion of neurotrophic factors, remodeling of neural circuits, restoration of the blood brain barrier, reduction of inflammatory infiltration and demyelination, and elevation of trophic factors. In addition to these mechanisms, we hypothesized that restored interhemispheric bilateral motor cortex connectivity might be an additional mechanism of functional recovery. In the present study, we have shown, with both MRI diffusion tensor imaging (DTI) and neuroanatomical tracing techniques using an adeno-associated virus (AAV) expressing GFP, that there was anatomical restoration of cortical interhemispheric connections through the corpus callosum after intravenous infusion of MSCs in a rat middle cerebral artery occlusion (MCAO) stroke model. Moreover, the degree of connectivity was greater in the MSC-treated group than in the vehicle-infused group. In accordance, both the thickness of corpus callosum and synaptic puncta in the contralateral (non-infarcted) motor cortex connected to the corpus callosum were greater in the MSC-treated group than in the vehicle group. Together, these results suggest that distinct preservation of interhemispheric cortical connections through corpus callosum was promoted by intravenous infusion of MSCs. This anatomical preservation of the motor cortex in the contralateral hemisphere may contribute to functional improvements following MSC therapy for cerebral stroke.

  DOI PubMed

• 脳小血管病と緊急血栓回収療法の臨床転帰との関係

  中崎公仁, 野中雅, 野村達史, 恩田敏之, 野中雅, 野村達史, 恩田敏之, 米増保之, 高橋明, 橋本祐治, 本田修, 佐々木祐典, 大坊雅彦, 本望修

  脳循環代謝(Web) ( 日本脳循環代謝学会 )  29 ( 1 ) 159 - 159  2017.11

  J-GLOBAL

• T1 Black Blood imagingによる椎骨動脈解離の経時的変化(Temporal Change of Signal Intensity of Intramural Hematoma in Patients with Acute Vertebral Artery Dissection Using T1-Weighted Black Blood Imaging)

  本田 修, 中崎 公仁, 佐々木 祐典, 野中 雅, 高橋 明, 橋本 祐治, 米増 保之, 野村 達史, 恩田 敏之, 大坊 雅彦

  CI研究 ( 日本脳神経CI学会 )  39 ( 2 ) 71 - 77  2017.11

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Industrial Property Rights 【 display / non-display 】

Industrial Property Rights 【 display / non-display 】

• 脳梗塞の治療用医薬

  本望 修, 中崎 公仁, 佐々木 祐典, 佐々木 優子, 岡 真一

  Patent

  J-GLOBAL

Awards 【 display / non-display 】

Awards 【 display / non-display 】

• 上原財団海外留学助成 リサーチフェローシップ

  2018.03  

• 脳卒中学会 最優秀口演賞

  2018.03  

• ヨーロッパ脳卒中学会 Young Investigator Award

  2009.05  

• 日本脳卒中学会 優秀ポスター賞

  2013.03  

Research Projects 【 display / non-display 】

Research Projects 【 display / non-display 】

• Restoration of cognitive impairment following intravenous infusion of mesenchymal stem cells in a rat model of cerebral small vessel disease

  Grant-in-Aid for Scientific Research (C)

  Project Year :

  2015.04

  -

  2018.03

   

  Nakazaki Masahito, HONMOU Osamu

  　View Summary

  Recently, intravenous mesenchymal stem cells (MSC) transplantation has been reported that greater therapeutic efficacy can be achieved via multiple mechanisms for various disorders. In this study, we aimed to investigate the effect of MSC transplantation in a rat model of vascular dementia by histological, MRI, and behavioral analysis. We used the Spontaneously hypertensive rats stroke-prone (SHRSP) with cognitive impairment. At week 21of age, transplantation of MSC was performed by intravenous injection. Compared with the vehicle group, the MSC infused group showed a significant improvement in cognitive function. The MRI demonstrated that the MSC infused group showed a significant reduction of brain atrophy compared to the vehicle group. The histological analysis also showed the volume of cerebral cortex and hippocampus were higher in MSC group than vehicle group. Collectively, intravenous infusion of MSCs might provide therapeutic efficacy in a rat model of vascular dementia.

• 脊髄損傷に対する骨髄間葉系幹細胞移植による、上位中枢神経における可塑性亢進の解析

  Grant-in-Aid for Scientific Research (C)

  Project Year :

  2018.04

  -

  2021.03

   

  岡 真一, 本望 修, 佐々木 祐典, 佐々木 優子, 中崎 公仁, 鈴木 淳平, 長濱 宏史

  　View Summary

  ラット脊髄損傷モデルに対して骨髄間葉系幹細胞(MSC)の経静脈投与することによって、中枢神経系にどのような神経回路の再構築(plasticity)が生じるかを検証するため、ラット胸髄Th9レベルの脊髄損傷モデルを作成し、動物用MRIでの撮像条件の最適化を検討した。8－9週齢のSDラットを購入し、ラット用脊髄損傷作成装置(IH-0400 Impactor; PSI社製)を用いて、安定した損傷部位を作成できることを確認した。その後、動物実験用高磁場MRI装置にて、脳、脊髄おける各種プロトコール（T1、T2、DTI等）での撮像を行い、撮像条件の調整を行い最適な撮像条件の検討を行った。撮像データの解析、特にDTIデータの解析を目的としてパーソナルコンピューターを別途購入し、専用のアプリケーションにて解析を実施した。 臨床においては、これまで当院にMSC治療を目的として入院した全ての脊髄損傷症例の脳、脊髄のMRIデータを検証し、入院時、投与直前、投与1ヶ月後、投与3ヶ月後、投与6ヶ月後の時点でのデータ解析を実施した。特に脳DTIデータを用いたコネクトーム解析では、標準化の方法、使用する脳atlasの選択、ROI間の接続強度の閾値設定など、症例間でばらつきのない均一な解析を実施するためのパラメータの設定を検討している。さらに、T1、T2などを用いて大脳皮質厚や体積などの構造学的なデータ解析も同時に実施し、コネクトーム解析で得られた結果との比較も実施している。また、脊髄MRIでのDTIでは、定量的な解析を目的に各種解析パラメータの検討を実施している。

• 実験的脊髄損傷後に生じる脳における可塑性の骨髄間葉系幹細胞による亢進作用の解析

  Grant-in-Aid for Scientific Research (C)

  Project Year :

  2018.04

  -

  2021.03

   

  長濱 宏史, 鈴木 淳平, 佐々木 祐典, 中崎 公仁, 本望 修

  　View Summary

  我々は、これまでに慢性期脊髄損傷動物モデルに対し、骨髄間葉系幹細胞 (mesenchymal stem cell: MSC)を経静脈的に移植し、運動機能の回復が得られることを報告した (Morita et al., 2016)。移植されたMSCは損傷部位に集積し、損傷局所のsproutingなどを誘導し機能改善に貢献していた。機能回復のメカニズムとして、損傷局所におけるsprouting などのメカニズムのみならず、損傷上位に位置する脊髄および脳において、MSC移植によってplasticityが亢進し、新しい神経回路が構築されると考えている。 さらに我々は、神経損傷動物モデルに対するMSC移植によって運動機能の改善を認めた群を、magnetic resonance imaging (MRI)を用いたdiffusion tensor tractograpy (DTT)による神経軸索イメージングと神経トレーサーによる神経解剖学的解析によって、MSC移植群では描出される神経線維数が、非MSC移植群に比べて有意に多いことを明らかにした。また、MSC移植群において、同部位におけるシナプス数は有意に多かった。これらの結果から、MSC移植によって神経線維が保護・強化されることで神経の可塑性を亢進させ、機能改善に貢献することを明らかにした (Nagahama et al., 2018)。 本研究では、動物モデルを用いて、MSC治療による脊髄損傷後の損傷局所、ならびに損傷上位脊髄および脳におけるplasticityの亢進によってどのように新しい神経回路が再構築され、運動機能の改善に寄与するかについて検討を行っている。以上より、補助金は適切に使用されている。

• 脊損に対する骨髄幹細胞移植における介在ニューロンにより再構築される神経回路の解析

  Grant-in-Aid for Scientific Research (C)

  Project Year :

  2018.04

  -

  2021.03

   

  廣田 亮介, 佐々木 祐典, 押切 勉, 森田 智慶, 寺島 嘉紀, 中崎 公仁, 岡 真一, 佐々木 優子, 山下 敏彦, 本望 修

  　View Summary

  「脊髄介在ニューロン」は、脊髄に存在する全ニューロンの約90%を占め、脊髄損傷後に神経回路ネットワークの再構築に関与することがわかっている。骨髄間葉系幹細胞（MSC）移植後では、皮質脊髄路の再生よりも早期に運動機能の回復が観察されることから、「脊髄介在ニューロン」が関わる神経回路ネットワークが機能的に再構築されることで、運動機能の回復に関与している可能性が高い。本研究では、この「脊髄介在ニューロン」が関与する神経回路ネットワークに対するMSCの治療効果を詳細に解明し、新たな治療法の開発へ展開することを目的としている。ラット(SD、オス)を全身麻酔下に、第9胸椎高位脊髄を圧挫することで脊髄損傷モデルラットを作成する。作製後1日目にラットから採集・培養したMSC（コントロール群にはMSCなしの培養液のみ）を経静脈的に投与し、コントロール群との間で行動解析を行っている。さらに、大脳に神経トレーサーを局所注入し、順行性に神経回路をトレースし、共焦点顕微鏡による観察およびneurolucida ソフトウェア(MBF)により、神経回路の高精度比較解析を行っている。

• 脳梗塞に対する骨髄幹細胞移植による脳のplasticity亢進による神経機能回復

  Grant-in-Aid for Scientific Research (C)

  Project Year :

  2017.04

  -

  2020.03

   

  鈴木 淳平, 佐々木 祐典, 長濱 宏史, 中崎 公仁, 本望 修

  　View Summary

  我々はこれまで，ラット脳梗塞モデル（middle cerebral artery occlusion: MCAO）へ骨髄間葉系幹細胞 (mesenchymal stem cell: MSC)を静脈内投与すると，治療効果が得られることを報告してきた．これまでに明らかにしてきた基礎研究の結果より，MSCによる治療効果のメカニズムは，①サイトカインによる神経栄養作用，②血液脳関門の安定化，③血管新生，④脱髄軸索の再有髄化，⑤脳のplasticityの賦活化など，と協奏的に発揮することによることが判明している． MSCの経静脈的移植により運動・感覚機能が回復する過程における脳のplasticityの変化を，動物用MRI（7Tesla）によるfunctional MRI（fMRI）を用いて解析した結果，脳のplasticityが変化することによって，脳内の神経回路のrewiringが賦活化され，神経機能の回復に寄与した可能性があると考えられた．MSCの経静脈的移植後に生ずる脳梗塞の運動機能の回復過程で，脳のplasticityの変化が重要な役割を果たしていることが示唆されたが，これまでのところ詳細な解明はなされていない． 本申請では，ラット脳梗塞モデル（MCAO）にMSCを経静脈的に投与した後に生じるこれらの脳のplasticityの変化を，Ex vivo MRI Diffusion Tensor Imagingおよび神経解剖学的および分子病理学的に解明し，新しい治療法の開発の基礎的なデータとすることを目的としている．現在までに，本研究費によってラット脳梗塞モデル（MCAO）を作成し，MSCの経静脈的投与を行った後に，動物実験用MRI装置（7Tesla）を用いてEx vivo Diffusion Tensor Imagingを撮像し，神経回路の解析および，神経トレーサーを用いた神経解剖学的解析や，免疫組織学的手法を用いて，大脳皮質のplasticityの変化を解析している．これらのデータはMSCの経静脈的投与を行った後の機能回復のメカニズムの解明に寄与すると思われる． 以上のように，補助金は補助条件に従って有効に使用されている．

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Presentations 【 display / non-display 】

Presentations 【 display / non-display 】

• 米国神経外傷学会 (Neurotrauma) Plenary session 招待講演

   [Invited]

• Systemic administration of mesenchymal stem cells inhibits reperfusion injury following intravenous rt-PA therapy for transient middle cerebral artery occlusion in rats

  日本脳卒中学会総会 2016