Pathomechanisms of developmental epileptic encephalopathy caused by CDKL5/CDKL3 mutations and development of the gene therapy
| Project/Area Number |
18K07784
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 52050:Embryonic medicine and pediatrics-related
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| Research Institution | The University of Tokyo |
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Principal Investigator |
Tanaka Teruyuki 東京大学, 大学院医学系研究科(医学部), 准教授 (10246647)
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| Project Period (FY) |
2018-04-01 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2020: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2019: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | CDKL5 / てんかん / 精神運動発達遅滞 / 遺伝子治療 / アデノ随伴ウイルス / ノックインマウス / ノックアウトマウス / アデノ随伴ウイルスベクター / 神経発達障害 / CDKL3 |
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| Outline of Final Research Achievements |
Several AAV vectors expressing hCDKL5 driven by the SynI promoter were generated to assess the efficiency of CDKL5 gene replacement by systemic AAV administration into the brains of Cdkl5 KO mice.
Cdkl5 kinase-dead knock-in mice were generated to model the CDKL5 missense mutations resulting in deletion of CDKL5 kinase activity. We identified abnormal phenotypes of the kinase-dead knock-in mice by comprehensive behavioral testing, impaired functional connectivity in the cerebral cortex by in vivo wide-range calcium imaging, and abnormalities in the gamma wave power spectrum by electroencephalography (EEG) analysis.
As a proof-of-concept, we generated Cdkl5 K42R(kinase-dead)-to-WT FLEx knock-in mice, a mouse model that allows CDKL5 gene replacement at any time by administration of tamoxifen.
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| Academic Significance and Societal Importance of the Research Achievements |
CDKL5欠損症に対するCDKL5遺伝子補充療法の前臨床試験をCdkl5 KOマウスを用いて行った。効果的な遺伝子補充に適したアデノ随伴ウイルス (AAV) ベクターを得るため数種類のAAVベクターを作製して効果を比較検討した。CDKL5遺伝子の病因変異のうちの、キナーゼ活性のみ欠失したCDKL5を発現するタイプのモデル動物として、CDKL5活性欠失マウスを作製し、その表現型を網羅的行動解析、in vivo大脳皮質広域カルシウムイメージング、脳波解析などにより多次元的に同定した。更に遺伝子補充療法の効果の概念実証のために、任意の時点で正常型マウスに変換可能なCdkl5機能喪失マウスを作製した。
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Report
(4 results)
Research Products
(16 results)
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[Presentation] CDKL5 controls postsynaptic localization of GluN2B-containing NMDA receptors in the hippocampus, and regulates seizure susceptibility, emotional behaviors, and memory2018
Author(s)
Teruyuki Tanaka, Kosuke Okuda, Shizuka Kobayashi, Takuto Murakami, Masahiro Fukaya, Keizo Takao, Aya Watanabe, Mai Hagiwara, Hiroyuki Sakagami, Masashi Mizuguchi, Tsuyoshi Miyakawa, Toshiya Manabe
Organizer
Cold Spring Harbor Asia - Latest Advances in Development & Function of Neuronal Circuits -
Related Report
Int'l Joint Research
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