Creation of viral vectors for sensitive detection of retinal ganglion cell damage and drug screening

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K22991
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0906:Surgery related to the biological and sensory functions and related fields
• Research Institution: Tohoku University
• Principal Investigator: Yabana Takeshi 東北大学, 大学病院, 特任助手 (30725213)
• Project Period (FY): 2020-09-11 – 2022-03-31
• Keywords: In vivo imaging / Ecel1 / Retinal ganglion cell / Glaucoma

Outline of Final Research Achievements

Using primers designed from the gene database, the promoter sequence of the Ecel1 gene on human DNA was amplified by PCR and ligated with an E. coli plasmid that has introduced the fluorescent protein in our previous studies. The plasmid was amplified by introducing it into E. coli and culturing, and the target sequence was confirmed by sequencing. Furthermore, the produced plasmid was packaged into AAV2 virus as the target viral vector. On the other hand, in order to adjust the newly introduced SLO camera, we prepared rats in which AAV2-CAG-EGFP was introduced into retinal ganglion cells (RGCs) by vitreous injection and confirmed that imaging of RGCs was possible by the device.

Free Research Field

緑内障

Academic Significance and Societal Importance of the Research Achievements

緑内障は、眼圧下降治療以外にエビデンスによって確立された治療法は未だ存在しない。その理由として緑内障は多因子疾患であり、眼圧非依存的な病因因子が複数存在すると推測される。これまでの基礎および臨床研究から、酸化ストレス、血流障害、軸索障害などで緑内障病態の主体である網膜神経節細胞が障害を受けることが示されている。
そこで、我々は網膜神経節細胞が軸索障害を受けた際に蛍光性を示す動物モデルを作成し、それを利用することで網膜神経節細胞の細胞死のメカニズムを解明し、さらには新しい緑内障治療の可能性を追究する。