研究者業績

八幡 直樹

ヤハタ ナオキ  (Naoki Yahata)

基本情報

所属
藤田医科大学 医学部 解剖学I 講師
学位
博士(理学)(大阪大学)

研究者番号
60450607
J-GLOBAL ID
200901060668151915
researchmap会員ID
5000050395

受賞

 1

論文

 14
  • Naoki Yahata, Hiroko Boda, Ryuji Hata
    Molecular therapy. Methods & clinical development 20 54-68 2021年3月12日  
    Various mitochondrial diseases, including mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes (MELAS), are associated with heteroplasmic mutations in mitochondrial DNA (mtDNA). Herein, we refined a previously generated G13513A mtDNA-targeted platinum transcription activator-like effector nuclease (G13513A-mpTALEN) to more efficiently manipulate mtDNA heteroplasmy in MELAS-induced pluripotent stem cells (iPSCs). Introduction of a nonconventional TALE array at position 6 in the mpTALEN monomer, which recognizes the sequence around the m.13513G>A position, improved the mpTALEN effect on the heteroplasmic shift. Furthermore, the reduced expression of the new Lv-mpTALEN(PKLB)/R-mpTALEN(PKR6C) pair by modifying codons in their expression vectors could suppress the reduction in the mtDNA copy number, which contributed to the rapid recovery of mtDNA in mpTALEN-applied iPSCs during subsequent culturing. Moreover, MELAS-iPSCs with a high proportion of G13513A mutant mtDNA showed unusual properties of spontaneous, embryoid body-mediated differentiation in vitro, which was relieved by decreasing the heteroplasmy level with G13513A-mpTALEN. Additionally, drug-inducible, myogenic differentiation 1 (MYOD)-transfected MELAS-iPSCs (MyoD-iPSCs) efficiently differentiated into myosin heavy chain-positive myocytes, with or without mutant mtDNA. Hence, heteroplasmic MyoD-iPSCs controlled by fine-tuned mpTALENs may contribute to a detailed analysis of the relationship between mutation load and cellular phenotypes in disease modeling.
  • Tsuneyoshi Seki, Motoi Kanagawa, Kazuhiro Kobayashi, Hisatomo Kowa, Naoki Yahata, Kei Maruyama, Nobuhisa Iwata, Haruhisa Inoue, Tatsushi Toda
    The Journal of biological chemistry 295(11) 3678-3691 2020年3月13日  査読有り
    Alzheimer's disease (AD) is the most common type of dementia, and its pathogenesis is associated with accumulation of β-amyloid (Aβ) peptides. Aβ is produced from amyloid precursor protein (APP) that is sequentially cleaved by β- and γ-secretases. Therefore, APP processing has been a target in therapeutic strategies for managing AD; however, no effective treatment of AD patients is currently available. Here, to identify endogenous factors that modulate Aβ production, we performed a gene microarray-based transcriptome analysis of neuronal cells derived from human induced pluripotent stem cells, because Aβ production in these cells changes during neuronal differentiation. We found that expression of the glycophosphatidylinositol-specific phospholipase D1 (GPLD1) gene is associated with these changes in Aβ production. GPLD1 overexpression in HEK293 cells increased the secretion of galectin 3-binding protein (GAL3BP), which suppressed Aβ production in an AD model, neuroglioma H4 cells. Mechanistically, GAL3BP suppressed Aβ production by directly interacting with APP and thereby inhibiting APP processing by β-secretase. Furthermore, we show that cells take up extracellularly added GAL3BP via endocytosis and that GAL3BP is localized in close proximity to APP in endosomes where amyloidogenic APP processing takes place. Taken together, our results indicate that GAL3BP may be a suitable target of AD-modifying drugs in future therapeutic strategies for managing AD.
  • 関 恒慶, 金川 基, 小林 千浩, 古和 久朋, 八幡 直樹, 丸山 敬, 岩田 修永, 井上 治久, 戸田 達史
    Dementia Japan 33(4) 516-516 2019年10月  査読有り
  • 関 恒慶, 金川 基, 小林 千浩, 古和 久朋, 八幡 直樹, 丸山 敬, 岩田 修永, 井上 治久, 戸田 達史
    日本生化学会大会プログラム・講演要旨集 92回 [2T17m-02] 2019年9月  査読有り
  • Naoki Yahata, Yuji Matsumoto, Minoru Omi, Naoki Yamamoto, Ryuji Hata
    SCIENTIFIC REPORTS 7 15557 2017年11月  査読有り
    Induced pluripotent stem cells (iPSCs) are suitable for studying mitochondrial diseases caused by mitochondrial DNA (mtDNA) mutations. Here, we generated iPSCs from a patient with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) with the m.13513G>A mutation. The patient's dermal fibroblasts were reprogrammed, and we established two iPSC clones with and without mutant mtDNA. Furthermore, we tried to decrease mutant mtDNA level in iPSCs using transcription activator-like effector nucleases (TALENs). We originally engineered platinum TALENs, which were transported into mitochondria, recognized the mtDNA sequence including the m.13513 position, and preferentially cleaved G13513A mutant mtDNA (G13513A-mpTALEN). The m.13513G>A heteroplasmy level in MELAS-iPSCs was decreased in the short term by transduction of G13513A-mpTALEN. Our data demonstrate that this mtDNA-targeted nuclease would be a powerful tool for changing the heteroplasmy level in heteroplasmic iPSCs, which could contribute to elucidation of the pathological mechanisms of mitochondrial diseases caused by mtDNA mutations.

MISC

 21

書籍等出版物

 2

講演・口頭発表等

 34

担当経験のある科目(授業)

 4

共同研究・競争的資金等の研究課題

 9

産業財産権

 1

その他

 2
  • 特になし
  • 変異ミトコンドリアDNAを標的としたTALEN(変異ミトコンドリアDNAの割合を改変する部位特的変異ヌクレアーゼ。Yahata et al. Sci Rep. 7:15557, 2017; Mol Ther Methods Clin Dev. 20:54-68, 2021) *本研究シーズに関する産学共同研究の問い合わせは藤田医科大学産学連携推進センター(fuji-san@fujita-hu.ac.jp)まで