Retinal optic nerve regeneration by human iPS cell-derived retinal ganglion cells and artificial sieve plate transplantation

2022 Fiscal Year Final Research Report

• Project/Area Number: 21K19548
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 56:Surgery related to the biological and sensory functions and related fields
• Research Institution: Tohoku University
• Principal Investigator: Nakazawa Toru 東北大学, 医学系研究科, 教授 (30361075)
• Co-Investigator(Kenkyū-buntansha): 小林 航 東北大学, 大学病院, 助教 (20646442) ; 佐藤 孝太 東北大学, 医学系研究科, 助教 (50732327) ; Sharma Parmanand 東北大学, 大学病院, 准教授 (80451623)
• Project Period (FY): 2021-07-09 – 2023-03-31
• Keywords: 網膜神経節細胞移植 / ヒトiPS細胞由来網膜神経節細胞

Outline of Final Research Achievements

The GFP fluorescent labeling to the Pou4F2 gene, a specific marker of retinal ganglion cells, was performed on human iPS cells by genome editing using Crispr/cas9 technology. The expression of GFP fluorescence was observed in the inner layer of the differentiated retinal organoids, and immunostaining of frozen sections showed co-staining of anti-GFP and anti-BRN3 antibodies. Human iPS-RGCs isolated from the three-dimensional retinal organoids were seeded into explant-cultured mouse retinas. The GFP-positive cells with neurites outgrowth were confirmed on the mouse retina by immunostaining on the fifth day after seeding. Human iPS-RGCs were injected into the vitreous of mice, but no GFP-positive cells were observed in the fundus of the eye after one week.

Free Research Field

眼科学

Academic Significance and Societal Importance of the Research Achievements

緑内障は、眼圧や環境要因に加えて遺伝要因が発症に関与する多因子疾患であり、網膜神経節細胞死により視野障害が生じる。中途失明原因の第一位であるが、眼圧下降治療以外に有効な治療がなく、眼圧下降治療に対して抵抗性を示す患者への新規治療法の開発は急務である。その一つとして失われた網膜神経節細胞を補充するヒトiPS細胞由来網膜神経節細胞(iPS-RGC)移植が挙げられる。本研究では、ヒトiPS-RGCをマウス網膜上へ播種した際に神経突起伸長を有して生着することが確認出来た。この研究結果からでは異種間の拒絶反応が生じない可能性があり、臨床応用する際にHLAを適合せずに移植出来る可能性が示唆された。