| Project/Area Number |
16K14582
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Neurochemistry/Neuropharmacology
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| Research Institution | Teikyo University |
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Principal Investigator |
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
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| Keywords | 骨髄間葉系幹細胞 / ゲノム編集 / microRNA / グルタチオン / 神経保護 / 神経細胞分化 / miR-96-5p / microRNA-96-5ｐ / 幹細胞分化 |
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| Outline of Final Research Achievements |
Improvement of stem cell function is necessary to succeed in autologous stem cell transplantation in the neurodegenerative diseases, because the stem cells are weakened in the patient of Alzheimer's disease and Parkinson's disease. Glutathione（GSH）protects the cells from oxidative stress. The intracellular GSH level is regulated by cysteine transporter EAAC1 in neuron and negatively regulated by miR-96-5p. Therefore we tried to make the bone marrow stem cells which lack miR-96 by genome editing. Bone marrow stem cells which have ability to generate neurons were prepared. CRISPR/Cas9 plasmids targeting miR-96 gene were constructed. The method for estimating the GSH level and the cell viability was established by using the flow cytometry. The efficiency of gene transfection was improved by using the electroporation technique. At present, we are estimating the viability of the miR-96 deficient neurons derived from the bone marrow stem cells which lack miR-96 by genome editing.
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| Academic Significance and Societal Importance of the Research Achievements |
神経変性疾患への自己骨髄幹細胞移植治療を実現するには、患者の幹細胞の劣化を改善する必要がある。細胞内のGSHの上昇は細胞に酸化ストレス抵抗性をもたらし、生存を促進する。本研究は神経細胞のGSHを増やす働きをするアミノ酸トランスポーターの発現を抑制するmiR96-5pをゲノム編集により欠損させ、GSH高発現の神経細胞を産生する骨髄幹細胞の作成を目指すものであり、得られた技術は再生医療の適応を神経変性疾患治療に広げることに貢献する。
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