| Project/Area Number |
18K15466
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 52020:Neurology-related
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| Research Institution | Juntendo University |
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Principal Investigator |
Yokota Mutsumi 順天堂大学, 大学院医学研究科, 助教 (10647415)
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| Project Period (FY) |
2018-04-01 – 2023-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
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| Keywords | ミトコンドリア / iPS細胞 / ドーパミン神経細胞 / PRKN / PRKN変異 / TH-GFP iPS細胞 / ドーパミン神経細胞死 / RNAシーケンス / ミトコンドリアスフェロイド化 / RNAシーケンス解析 / CLEM / CRISPR/Cas9 / GFP標識 / PARK2 iPS細胞 / 超微形態学的解析 |
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| Outline of Final Research Achievements |
Parkinson’s disease with PRKN mutations is characterized by the preferential degeneration of dopaminergic neurons in the substantia nigra pars compacta. However, ultrastructural changes of mitochondria specifically in dopaminergic neurons derived from iPSC have rarely been analyzed. I established tyrosine hydroxylase reporter (TH-GFP) iPSC lines from a PD patient with a PRKN mutation to perform correlative light-electron microscopy analysis and live cell imaging in GFP-expressing dopaminergic neurons. In addition, a proximity ligation assay and mitochondrial calcium imaging using TH-GFP iPSCs showed that ER-mitochondrial contact sites were significantly reduced in PRKN-mutant patient dopaminergic neurons compared to the control. This study using TH-GFP iPSC lines would contribute to further understanding of the mechanisms of dopaminergic neuron degeneration in patients with PRKN mutations.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究により、ドーパミン神経細胞においてGFPを発現するパーキンソン病患者iPS細胞株が得られた。これまでTH遺伝子にレポーターをノックインしたiPS細胞株は健常者についてのみ報告されており、PRKN変異患者TH-GFP iPS細胞の樹立については未だ報告がなかった。TH-GFP iPS細胞株 を用いることによりドーパミン神経細胞特異的なオルガネラの形態学的解析やライブイメージングが可能となり、パーキンソン病に至るオルガネラの形態・機能的特徴を見出す可能性があるため本研究結果は重要な成果である。
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