Gene therapy for glaucoma by stimulating neuroprotection and regeneration

• Project/Area Number: 20K09820
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 56060:Ophthalmology-related
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• Principal Investigator: KIMURA Atsuko 公益財団法人東京都医学総合研究所, 疾患制御研究分野, 研究員 (60569143)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 緑内障 / TrkB / 神経保護 / AAV / RGC / 遺伝子治療 / 軸索再生 / 神経再生

Outline of Research at the Start

緑内障は視神経軸索が障害を受け、その細胞体であるRGCが死に至ることで視野障害が起きる神経変性疾患である。近年では軸索終末や樹状突起の退縮が起こり、神経回路が切断されることが発症原因とも考えられている。本研究では、軸索と樹状突起を再生させることで、神経回路を再構築し、一度低下した視機能が回復するか検討する。これには独自に作製したBDNFの受容体であるTrkBの常時活性型ウイルスベクター（AAV2-CA-TrkB）を活用し、視神経挫滅モデルにおいてTrkBシグナル増強による軸索および樹状突起の保護・再生のメカニズムを追求し、AAV2-CA-TrkBによる遺伝子治療の有効性を検証する。

Outline of Final Research Achievements

We have made a constitutive-active TrkB molecule, which is artificially modified receptor for BDNF. Gene therapy of constitutive-active TrkB using adeno associated virus induced neuroprotection and axon regeneration in mice after optic nerve crush. However, the molecular mechanism is unknown. In this study, we have investigated gene expression pattern in RGC using RNA-seq after overexpression of constitutive-active TrkB. We found that constitutive-active TrkB changed the expression levels in many genes comparted with control. In addition, neuroprotective effect was found in hypertension glaucoma model mice by gene therapy of constitutive-active TrkB.

Academic Significance and Societal Importance of the Research Achievements

F-iTrkBのウイルスベクターによる神経保護・再生の際に、変動する遺伝子群をRNA-seqによって網羅的に解析することで、細胞内におけるシグナル応答についてより深い知見を得ることができた。得られたデータを活用して、今後はより効果的な遺伝治療の開発につながることを期待したい。

Research Products

• [Journal Article] Effects of constitutively active K-Ras on axon regeneration after optic nerve injury (2023)
  • Author(s): Kiyota Naoki、Namekata Kazuhiko、Nishijima Euido、Guo Xiaoli、Kimura Atsuko、Harada Chikako、Nakazawa Toru、Harada Takayuki
  • Journal Title: Neuroscience Letters Volume: 799 Pages: 137124-137124
  • DOI: 10.1016/j.neulet.2023.137124
  • Peer Reviewed / Open Access
• [Journal Article] Vision protection and robust axon regeneration in glaucoma models by membrane-associated Trk receptors (2023)
  • Author(s): Nishijima Euido、Honda Sari、Kitamura Yuta、Namekata Kazuhiko、Kimura Atsuko、Guo Xiaoli、Azuchi Yuriko、Harada Chikako、Murakami Akira、Matsuda Akira、Nakano Tadashi、Parada Luis F.、Harada Takayuki
  • Journal Title: Molecular Therapy Volume: 31 Issue: 3 Pages: 810-824
  • DOI: 10.1016/j.ymthe.2022.11.018
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ASK1 signaling regulates phase-specific glial interactions during neuroinflammation (2022)
  • Author(s): Guo Xiaoli、Kimura Atsuko、Namekata Kazuhiko、Harada Chikako、Arai Nobutaka、Takeda Kohsuke、Ichijo Hidenori、Harada Takayuki
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 119 Issue: 6
  • DOI: 10.1073/pnas.2103812119
  • Peer Reviewed / Open Access
• [Journal Article] Effects of lighting environment on the degeneration of retinal ganglion cells in glutamate/aspartate transporter deficient mice, a mouse model of normal tension glaucoma (2022)
  • Author(s): Ohashi Tsutomu、Namekata Kazuhiko、Guo Xiaoli、Kimura Atsuko、Harada Chikako、Harada Takayuki
  • Journal Title: Biochemistry and Biophysics Reports Volume: 29 Pages: 101197-101197
  • DOI: 10.1016/j.bbrep.2021.101197
  • Peer Reviewed / Open Access
• [Journal Article] リパスジル点眼は視神経損傷後の成体マウスにおいて神経保護と軸索再生を促進する (2021)
  • Author(s): 西島義道、行方和彦、木村敦子、郭 暁麗、原田知加子、野呂隆彦、中野 匡、原田高幸
  • Journal Title: 日本眼科学会雑誌 Volume: 125 Pages: 82-82
• [Journal Article] Roles of the DOCK-D family proteins in a mouse model of neuroinflammation (2020)
  • Author(s): Namekata Kazuhiko、Guo Xiaoli、Kimura Atsuko、Azuchi Yuriko、Kitamura Yuta、Harada Chikako、Harada Takayuki
  • Journal Title: Journal of Biological Chemistry Volume: in press Issue: 19 Pages: 6710-6720
  • DOI: 10.1074/jbc.ra119.010438
  • Peer Reviewed / Open Access
• [Journal Article] Topical ripasudil stimulates neuroprotection and axon regeneration in adult mice following optic nerve injury (2020)
  • Author(s): Nishijima Euido、Namekata Kazuhiko、Kimura Atsuko、Guo Xiaoli、Harada Chikako、Noro Takahiko、Nakano Tadashi、Harada Takayuki
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 15709-15709
  • DOI: 10.1038/s41598-020-72748-3
  • Peer Reviewed / Open Access
• [Presentation] Gene therapy with modified TrkB induces neuroprotection and axon regeneration (2021)
  • Author(s): Atsuko Kimura, Euido Nishijima, Yuta Kitamura, Sari Honda, Xiaoli Guo, Chikako Harada, Kazuhiko Namekata, Takayuki Harada
  • Organizer: 第44回日本神経科学大会
  • Int'l Joint Research
• [Presentation] Microglial DOCK8 and neurodegeneration (2021)
  • Author(s): Atsuko Kimura, Kazuhiko Namekata, Xiaoli Guo, Nobutaka Arai, Chikako Harada, Takayuki Harada
  • Organizer: The 4th International Conference on Applied Biochemistry and Biotechnology
  • Int'l Joint Research / Invited
• [Presentation] AAV-mediated delivery of constitutively active TrkB promotes CNS axon regeneration and neuroprotection (2021)
  • Author(s): Atsuko Kimura, Euido Nishijima, Yuta Kitamura, Sari Honda, Xiaoli Guo, Chikako Harada, Kazuhiko Namekata, Takayuki Harada
  • Organizer: Neuroscience 2021
  • Int'l Joint Research
• [Presentation] 活性型Rasの遺伝子治療による視神経保護と再生への影響 (2020)
  • Author(s): 金義道、本田紗里、北村裕太、郭暁麗、木村敦子、行方和彦、中野匡、原田高幸
  • Organizer: 第１２４回日本眼科学会総会
• [Presentation] 活性型TrkBを用いた遺伝子治療は上丘における網膜神経節細胞の軸索再生を促進する (2020)
  • Author(s): 本田紗里、金義道、北村裕太、郭暁麗、木村敦子、行方和彦、松田彰、村上晶、原田高幸
  • Organizer: 第１２４回日本眼科学会総会
• [Presentation] TrkB欠損マウスにおける網膜神経節細胞の変性過程の検討 (2020)
  • Author(s): 北村裕太、安土ゆり子、行方和彦、木村敦子、郭暁麗、山本修一、原田高幸
  • Organizer: 第１２４回日本眼科学会総会
• [Presentation] 失明からの視機能回復に向けた新規遺伝子治療ベクターの開発 (2020)
  • Author(s): 金義道、行方和彦、本田紗里、木村敦子、北村裕太、中野匡、原田高幸
  • Organizer: 第１９回日本再生医療学会総会
• [Presentation] GLAST欠損マウスによる網膜神経節細胞樹状突起の解析 (2020)
  • Author(s): 北村裕太、行方和彦、木村敦子、郭暁麗、山本修一、原田高幸
  • Organizer: 第３１回日本緑内障学会
• [Presentation] TrkB欠損マウスにおける網膜神経節細胞の変性過程の検討 (2020)
  • Author(s): 北村裕太、安土ゆり子、行方和彦、木村敦子、郭暁麗、山本修一、原田高幸
  • Organizer: 第７４回日本臨床眼科学会
• [Remarks] 研究室HP
  • URL: https://www.igakuken.or.jp/retina/