Construction of nanocapsule targeting mutated mitochondria for gene therapy

• Project/Area Number: 17H02094
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biomedical engineering/Biomaterial science and engineering
• Research Institution: Hokkaido University
• Principal Investigator: Yamada Yuma 北海道大学, 薬学研究院, 准教授 (60451431)
• Co-Investigator(Kenkyū-buntansha): 田村 篤志 東京医科歯科大学, 生体材料工学研究所, 准教授 (80631150)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000); Fiscal Year 2019: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2018: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2017: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
• Keywords: 薬物送達システム / ミトコンドリア / 薬物送達

Outline of Final Research Achievements

In this study, we validated a mitochondrial gene therapeutic strategy using fibroblasts from a mitochondrial disease patient by the mitochondrial delivery of therapeutic nucleic acids. The treatment involved the use of a liposome-based carrier (a MITO-Porter) for delivering therapeutic nucleic acids to mitochondria via membrane fusion. The results confirmed that the mitochondrial transfection of therapeutic nucleic acids by the MITO-Porter system resulted in a decrease in the levels of mutated mitochondria of diseased cells. An evaluation of mitochondrial respiratory activity by respirometry also showed that transfection using the MITO-Porter resulted in an increase in maximal mitochondrial respiratory activity in the diseased cells. These results support the conclusion that the mitochondrial delivery of therapeutic nucleic acids represents a viable strategy for mitochondrial gene therapy.

Academic Significance and Societal Importance of the Research Achievements

難病指定されているミトコンドリア病などミトコンドリア機能異常が関連する疾患の多くはミトコンドリア遺伝子の変異・欠損が原因であることが報告さているが、現在の治療法はビタミン剤などを継続的に投与し、ミトコンドリア機能を一時的に補助する対症療法が主流である。本研究では、ミトコンドリア標的型ナノカプセル【MITO-Porter】を用いて、患者由来細胞に治療用核酸を送達する「ミトコンドリアを標的とした遺伝治療戦略」の実証に成功した。ミトコンドリアを標的とした遺伝子治療は、これまでの治療法に代わる難病の根本治療につながるため医療に大きなインパクトを与える。

Research Products

• [Journal Article] The optimization of cancer photodynamic therapy by utilization of a pi-extended porphyrin-type photosensitizer in combination with MITO-Porter (2020)
  • Author(s): Satrialdi, Munechika R, Biju V, Takano Y, Harashima H, Yamada Y.
  • Journal Title: ChemCommun. Volume: 56 Issue: 7 Pages: 1145-1148
  • DOI: 10.1039/c9cc08563g
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Development of a nanoparticle that releases nucleic acids in response to a mitochondrial environment. (2020)
  • Author(s): Yamada Y, Fukuda Y, Sasaki D, Maruyama M, Harashima H
  • Journal Title: Mitochondrion Volume: 52 Pages: 67-74
  • DOI: 10.1016/j.mito.2020.02.009
  • Peer Reviewed
• [Journal Article] Validation of gene therapy for mutant mitochondria by delivering mitochondrial RNA using a MITO-Porter, a liposome-based nano device (2020)
  • Author(s): Kawamura E, Maruyama M, Abe J, Sudo A, Takeda A, Takada S, Yokota T, Kinugawa S, Harashima H, Yamada Y
  • Journal Title: Mol. Ther. - Nucleic Acids Volume: - Pages: 687-698
  • DOI: 10.1016/j.omtn.2020.04.004
  • Peer Reviewed / Open Access
• [Journal Article] Validation of a mitochondrial RNA therapeutic strategy using fibroblasts from a Leigh syndrome patient with a mutation in the mitochondrial ND3 gene (2020)
  • Author(s): Yamada Y, Somiya K, Miyauchi A, Osaka H, Harashima H
  • Journal Title: Sci. Rep Volume: - Issue: 1 Pages: 7511-7511
  • DOI: 10.1038/s41598-020-64322-8
  • Peer Reviewed / Open Access
• [Journal Article] A Nanocarrier System for Mitochondrial Delivery Targeted to a Pancreatic Beta Cell (2019)
  • Author(s): Yamada Yuma
  • Journal Title: YAKUGAKU ZASSHI Volume: 139 Issue: 1 Pages: 41-45
  • DOI: 10.1248/yakushi.18-00163-1
  • NAID: 130007541687
  • ISSN: 0031-6903, 1347-5231
  • Year and Date: 2019-01-01
  • Peer Reviewed
• [Journal Article] Enhanced autophagy induction via the mitochondrial delivery of methylated β-cyclodextrin-threaded polyrotaxanes using a MITO-Porter (2019)
  • Author(s): Yamada Yuma、Daikuhara Shinnosuke、Tamura Atsushi、Nishida Kei、Yui Nobuhiko、Harashima Hideyoshi
  • Journal Title: Chemical Communications Volume: 55 Issue: 50 Pages: 7203-7206
  • DOI: 10.1039/c9cc03272j
  • Peer Reviewed
• [Journal Article] A mitochondrial delivery system using liposome-based nanocarriers that target myoblast cells (2019)
  • Author(s): Katayama Takashi、Kinugawa Shintaro、Takada Shingo、Furihata Takaaki、Fukushima Arata、Yokota Takashi、Anzai Toshihisa、Hibino Mitsue、Harashima Hideyoshi、Yamada Yuma
  • Journal Title: Mitochondrion Volume: 49 Pages: 66-72
  • DOI: 10.1016/j.mito.2019.07.005
  • Peer Reviewed / Open Access
• [Journal Article] Targeted mitochondrial delivery of antisense RNA-containing nanoparticles by a MITO-Porter for safe and efficient mitochondrial gene silencing. (2019)
  • Author(s): 396.Kawamura E, Hibino M, Harashima H, Yamada Y.
  • Journal Title: Mitochondrion. Volume: 28 Pages: 178-188
  • DOI: 10.1016/j.mito.2019.08.004
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] ミトコンドリアを標的とする遺伝子治療用ナノカプセルの創製 (2019)
  • Author(s): 山田勇磨
  • Journal Title: 日本核酸医薬学会会誌 Volume: - Pages: 12-21
  • NAID: 40022467016
  • Peer Reviewed
• [Journal Article] Innovative nanotechnologies for enhancing nucleic acids/gene therapy: Controlling intracellular trafficking to targeted biodistribution (2019)
  • Author(s): Nakamura Takashi, Yamada Yuma, Sato Yusuke, Khalil A Ikramy, Harashima Hideyoshi
  • Journal Title: Biomaterials Volume: 218 Pages: 119329-119329
  • DOI: 10.1016/j.biomaterials.2019.119329
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The use of a microfluidic device to encapsulate a poorly water-soluble drug CoQ10 in lipid nanoparticles and an attempt to regulate intracellular trafficking to reach mitochondria (2019)
  • Author(s): Hibino Mitsue、Yamada Yuma、Fujishita Naoki、Sato Yusuke、Maeki Masatoshi、Tokeshi Manabu、Harashima Hideyoshi
  • Journal Title: Journal of Pharmaceutical Sciences Volume: 印刷中 Issue: 8 Pages: 2668-2676
  • DOI: 10.1016/j.xphs.2019.04.001
  • Peer Reviewed
• [Journal Article] ミトコンドリア標的型DDSを基盤とした革新的ナノ医薬品の創製 (2019)
  • Author(s): 山田勇磨
  • Journal Title: ファルマシア Volume: 55 Pages: 330-334
  • NAID: 130007621948
  • Peer Reviewed
• [Journal Article] Future of human mitochondrial DNA editing technologies (2019)
  • Author(s): Verechshagina N, Nikitchina N, Yamada Y, Harashima H, Tanaka M, Orishchenko K, Mazunin I
  • Journal Title: Mitochondrial DNA Part A Mapp Seq Anal Volume: 30 Issue: 2 Pages: 214-221
  • DOI: 10.1080/24701394.2018.1472773
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ミトコンドリアDDSが拓く新しいミトコンドリア病治療戦略 (2019)
  • Author(s): 山田勇磨、原島秀吉
  • Journal Title: 小児科診療 Volume: 823 Pages: 523-528
• [Journal Article] ミトコンドリアへの分子送達技術 (2018)
  • Author(s): 山田勇磨、原島秀吉
  • Journal Title: BIO Clinica Volume: 33 Pages: 637-640
• [Journal Article] Optimization of siRNA delivery to target sites: issues and future directions (2018)
  • Author(s): Khalil Ikramy A.、Yamada Yuma、Harashima Hideyoshi
  • Journal Title: Expert Opinion on Drug Delivery Volume: 15 Issue: 11 Pages: 1053-1065
  • DOI: 10.1080/17425247.2018.1520836
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] In Vivo Transgene Expression in the Pancreas by the Intraductal Injection of Naked Plasmid DNA. (2018)
  • Author(s): Yamada Y, Tabata M, Abe J, Nomura M, Harashima H.
  • Journal Title: J Pharm Sci. Volume: 107 Issue: 2 Pages: 647-653
  • DOI: 10.1016/j.xphs.2017.09.021
  • NAID: 120006558999
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Mitochondrial transgene expression via an artificial mitochondrial DNA vector in cells from a patient with a mitochondrial disease (2018)
  • Author(s): Ishikawa Takuya、Somiya Kana、Munechika Reina、Harashima Hideyoshi、Yamada Yuma
  • Journal Title: J Control Res Volume: 274 Pages: 109-117
  • DOI: 10.1016/j.jconrel.2018.02.005
  • NAID: 120006595504
  • Peer Reviewed
• [Journal Article] Mitochondrial Delivery of Doxorubicin Using MITO-Porter Kills Drug-Resistant Renal Cancer Cells via Mitochondrial Toxicity (2017)
  • Author(s): Yamada Yuma、Munechika Reina、Kawamura Eriko、Sakurai Yu、Sato Yusuke、Harashima Hideyoshi
  • Journal Title: J Pharm Sci Volume: 106 Issue: 9 Pages: 2428-2437
  • DOI: 10.1016/j.xphs.2017.04.058
  • Peer Reviewed
• [Journal Article] Packaging of the Coenzyme Q₁₀ into a Liposome for Mitochondrial Delivery and the Intracellular Observation in Patient Derived Mitochondrial Disease Cells (2017)
  • Author(s): Yamada Yuma、Burger Laila、Kawamura Eriko、Harashima Hideyoshi
  • Journal Title: Biological and Pharmaceutical Bulletin Volume: 40 Issue: 12 Pages: 2183-2190
  • DOI: 10.1248/bpb.b17-00609
  • NAID: 130006235437
  • ISSN: 0918-6158, 1347-5215
  • Peer Reviewed
• [Journal Article] Optical control of mitochondrial reductive reactions in living cells using an electron donor?acceptor linked molecule (2017)
  • Author(s): Takano Yuta、Munechika Reina、Biju Vasudevanpillai、Harashima Hideyoshi、Imahori Hiroshi、Yamada Yuma
  • Journal Title: Nanoscale Volume: 9 Issue: 47 Pages: 18690-18698
  • DOI: 10.1039/c7nr06310e
  • NAID: 120006545088
  • Peer Reviewed / Open Access
• [Journal Article] Validation of the use of an artificial mitochondrial reporter DNA vector containing a Cytomegalovirus promoter for mitochondrial transgene expression. (2017)
  • Author(s): Yamada Y, Ishikawa T, Harashima H.
  • Journal Title: Biomaterials. Volume: 136 Pages: 56-66
  • DOI: 10.1016/j.biomaterials.2017.05.016
  • NAID: 120006716305
  • Peer Reviewed / Open Access
• [Journal Article] ミトコンドリアを標的とする核酸医薬研究 (2017)
  • Author(s): 山田勇磨
  • Journal Title: 医学のあゆみ Volume: 262 Pages: 142-146
• [Presentation] MITO-Porter, liposomal mitochondrial delivery system: Toward Mitochondrial Nanomedicine. (2019)
  • Author(s): Yamada Y, Harashima H
  • Organizer: Liposome Research Days 2019
  • Int'l Joint Research / Invited
• [Presentation] MITO-Porter therapy to open innovative therapies targeting mitochondria. (2019)
  • Author(s): Yamada Y
  • Organizer: 16th ASMRM & 19th J-mit.
  • Int'l Joint Research / Invited
• [Presentation] ミトコンドリア標的型ナノＤＤＳを基盤とした遺伝子・細胞治療の実装 (2019)
  • Author(s): 山田 勇磨
  • Organizer: 第12回 CHEMBIOハイブリッドレクチャー
  • Invited
• [Presentation] ミトコンドリアDDSが描くナノ医療の未来予想図～リポ化プロスタグランジン製剤の話題を含めて～ (2019)
  • Author(s): 山田 勇磨
  • Organizer: 第12回 北海道小児オータムセミナー
  • Invited
• [Presentation] Mitochondrial transgene expression by the mitochondrial delivery of an artificial mitochondrial DNA vector using a MITO-Porter system (2019)
  • Author(s): Yamada Y, Ishikawa T, Harashima H
  • Organizer: 2019 Annual Meeting & Exposition of the Controlled Release Society
• [Presentation] Validation of mitochondrial RNA therapy for mitochondrial diseases using Liposome-based nano device (2019)
  • Author(s): Somiya K, Yamada Y, Miyauchi A, Osaka H, Harashima H
  • Organizer: Liposome Research Days 2019
• [Presentation] Development of ribosomal RNA-loaded nano device for mitochondrial delivery to achieve mitochondrial deafness related gene therapy (2019)
  • Author(s): Maruyama M, Yamada Y, Kita T, Kitajiri S, Usami S, Harashima H
  • Organizer: Liposome Research Days 2019
• [Presentation] Mitochondrial Nano DDS Toward Innovative Medicine and Therapy. (2018)
  • Author(s): Yamada Y
  • Organizer: HIGO Program Cutting edge Seminar.
  • Invited
• [Presentation] ミトコンドリアDDSの使い方 (2018)
  • Author(s): 山田勇磨
  • Organizer: 第9回 フリーラジカルスクール
  • Invited
• [Presentation] ミトコンドリア機能を制御するナノカプセルの構築および疾患治療に向けた試み (2018)
  • Author(s): 山田 勇磨、原島秀吉
  • Organizer: 第91回 日本生化学大会
  • Invited
• [Presentation] Mitochondrial DDS opens emerging therapies and novel strategy of new drug development for mitochondrial disorders. (2018)
  • Author(s): Yamada Y, Harashima H
  • Organizer: KSAP annual convention 2018
  • Int'l Joint Research / Invited
• [Presentation] ミトコンドリア標的型DDSを基盤としたナノ医療の創出を目指して (2018)
  • Author(s): 山田 勇磨、原島秀吉
  • Organizer: 第18回 日本ミトコンドリア学会年会
  • Invited
• [Presentation] The MITO-Porter integrates a mitochondrial drug delivery system with a variety of other current scientific concepts with the goal of developing new innovative technologies and medicines. (2018)
  • Author(s): Yamada Y, Harashima H
  • Organizer: The 19th RIES-HOKUDAI International Symposium.
  • Int'l Joint Research / Invited
• [Presentation] ミトコンドリアDDSが拓く新しいミトコンドリア病治療戦略 (2018)
  • Author(s): 山田 勇磨、原島秀吉
  • Organizer: 第60回日本小児神経学会学術集会
  • Invited
• [Presentation] 遺伝子・細胞治療に用いるミトコンドリアDDS (2018)
  • Author(s): 山田 勇磨、原島秀吉
  • Organizer: 第34回日本DDS学会
  • Invited
• [Presentation] ミトコンドリアを標的とする遺伝子治療用ナノカプセルの創製 (2018)
  • Author(s): 山田 勇磨
  • Organizer: 日本核酸医薬学会第4回年会
  • Invited
• [Presentation] Mitochondrial DDS for Gene & Cell Therapy. (2018)
  • Author(s): Yamada Y, Harashima H
  • Organizer: Cadiovascular and Metabolic Week 2018
  • Int'l Joint Research / Invited
• [Presentation] ND3変異細胞を対象とした治療用mRNA送達によるミトコンドリア遺伝子治療戦略の検証 (2018)
  • Author(s): 宗宮加奈、山田勇磨、河村恵理子、宮内彰彦、小坂仁、原島秀吉
  • Organizer: 日本薬学会北海道支部第145回例会
• [Presentation] ミトコンドリア標的型DDSを用いたミトコンドリア病に対する遺伝子治療戦略の検証 (2018)
  • Author(s): 宗宮加奈、山田勇磨、宮内彰彦、小坂仁、原島秀吉
  • Organizer: 第18回 日本ミトコンドリア学会年会
• [Presentation] ミトコンドリア機能を制御するナノカプセルの開発 (2018)
  • Author(s): 山田勇磨
  • Organizer: 第6回ミトコンドリア機能研究会
  • Invited
• [Presentation] 膵島ミトコンドリアを標的とした分子送達技術の開発および糖尿病治療への展開 (2018)
  • Author(s): 山田勇磨
  • Organizer: 日本薬学会 第138年会
  • Invited
• [Presentation] ミトコンドリア病の遺伝子改変治療 (2018)
  • Author(s): 山田 勇磨
  • Organizer: H29 年度 村山班ミトコンドリア病診断・診療マニュアル作成委員拡大合宿
  • Invited
• [Presentation] 治療用RNA搭載ミトコンドリア標的型リポソームによる遺伝子治療戦略の検証 (2018)
  • Author(s): 宗宮加奈、山田勇磨、河村恵理子、宮内彰彦、小坂仁、原島秀吉
  • Organizer: 日本薬学会 第138年会
• [Presentation] ミトコンドリアを診る・操る・治すDrug Delivery System (2017)
  • Author(s): 山田勇磨
  • Organizer: 第2回先端ケミカルバイオロジー研究会
  • Invited
• [Presentation] ミトコンドリアＤＤＳの開発と医療分野への展開 (2017)
  • Author(s): 山田勇磨
  • Organizer: 第11回 次世代を担う若手医療薬科学シンポジウム
  • Invited
• [Presentation] 癌細胞ミトコンドリアを標的とした光殺細胞効果の検証 (2017)
  • Author(s): 山田 勇磨、高野勇太、宗近玲那、福田豊、今堀博、原島秀吉
  • Organizer: 第70回日本酸化ストレス学会学術集会
• [Presentation] ミトコンドリアDDSを基盤とした遺伝子・細胞治療への展開 (2017)
  • Author(s): 山田 勇磨、原島秀吉
  • Organizer: 第33回日本DDS学会
  • Invited
• [Presentation] Validation of mitochondrial gene therapy using a MITO-Porter (2017)
  • Author(s): Yamada Y, Harashima H
  • Organizer: 8th SFRR-Asia & 14th ASMRM
  • Int'l Joint Research / Invited
• [Presentation] ミトコンドリア標的型ナノカプセルMITO-Porterを基盤とした遺伝子治療への挑戦 (2017)
  • Author(s): 山田 勇磨、原島秀吉
  • Organizer: 第59回日本先天代謝異常学会
  • Invited
• [Presentation] Nano-drug delivery system for mitochondrial gene therapy (2017)
  • Author(s): Yamada Y, Harashima H
  • Organizer: The 18th International Union of Materials Research Societies, International Conference in Asia (IUMRS-ICA) 2017
  • Int'l Joint Research / Invited
• [Presentation] PIポリアミドによる病因性ミトコンドリアDNA変異関連疾患の治療 (2017)
  • Author(s): 越川 信子、渡部 隆義、篠崎 喜脩、高取 敦志、安井 七海、リン ジェイソン、山田 勇磨、原島 秀吉、永瀬 浩喜
  • Organizer: 2017年度 生命科学系学会合同年次大会
  • Invited
• [Presentation] VALIDATION OF MITOCHONDRIAL GENE THERAPY BY MITOCHONDRIAL DELIVERY OF THERAPEUTIC NUCLEIC ACIDS (2017)
  • Author(s): Yamada Y, Kawamura E, Harashima H
  • Organizer: EUROMIT2017
  • Int'l Joint Research
• [Presentation] ミトコンドリア外来遺伝子発現を可能とするDDSの開発 (2017)
  • Author(s): 山田勇磨、石川卓哉、原島秀吉
  • Organizer: 日本薬剤学会第32年会
• [Presentation] ミトコンドリアマトリクス環境応答性を有する核酸ナノ粒子の設計 (2017)
  • Author(s): 山田 勇磨、福田豊、原島秀吉
  • Organizer: 日本核酸医薬学会第3回年会
• [Remarks] 北海道大学 大学院薬学研究院 薬剤分子設計学研究室HP
  • URL: https://www.pharm.hokudai.ac.jp/yakusetu/
• [Remarks] 北海道大学 大学院薬学研究院 HP
  • URL: https://www.pharm.hokudai.ac.jp/
• [Remarks] ルカ・サイエンス
  • URL: https://luca-science.com/
• [Remarks] 7SEAS PROJECT HP
  • URL: https://7sp.life/
• [Patent(Industrial Property Rights)] ミトコンドリアリボソームＲＮＡ変異検出用プライマーＤＮＡセットを含むキット、ミトコンドリアリボソームＲＮＡ発現用核酸、前記核酸を封入してなる脂質膜構造体及びそれらの利用 (2019)
  • Inventor(s): 山田勇磨, 原島秀吉, 丸山美菜子
  • Industrial Property Rights Holder: ルカ・サイエンス
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-089566
  • Filing Date: 2019
• [Patent(Industrial Property Rights)] ミトコンドリア内でタンパク質を発現させるための核酸、前記核酸を封入した脂質膜構造体及びそれらの利用 (2019)
  • Inventor(s): 原島秀吉, 山田勇磨, 宗宮加奈, 小坂仁
  • Industrial Property Rights Holder: 原島秀吉, 山田勇磨, 宗宮加奈, 小坂仁
  • Industrial Property Rights Type: 特許
  • Filing Date: 2019
  • Overseas
• [Patent(Industrial Property Rights)] ミトコンドリア内でタンパク質を発現させるための核酸、前記核酸を封入した脂質膜構造体及びそれらの利用 (2018)
  • Inventor(s): 原島秀吉、山田勇磨、宗宮加奈、小坂仁
  • Industrial Property Rights Holder: 原島秀吉、山田勇磨、宗宮加奈、小坂仁
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2018-031349
  • Filing Date: 2018