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Satoshi Inagaki, Kazuhide Kawase, Michinori Funato, Kazuki Ohuchi, Shiori Ando, Arisu Sato, Wataru Morozumi, Masamitsu Shimazawa, Daisuke Iejima, Takeshi Iwata, Tetsuya Yamamoto, Hideo Kaneko, Hideaki Hara; The oligodendrocyte derived from familial glaucoma patients-iPSCs decreases neuroprotective action for retinal ganglion cells. Invest. Ophthalmol. Vis. Sci. 2018;59(9):3751. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Glaucoma is characterized by a progressive loss of retinal ganglion cells (RGCs) with axonal degeneration, suggesting that axonal degeneration precede RGCs death. Structurally, RGCs axons are surrounded by oligodendrocytes (OLs). Therefore, our purpose is to elucidate whether OLs would contribute to axonal degeneration and RGCs death using patient RGCs or OLs culture system using induced pluripotent stem cells (iPSCs) derived from patients who have E50K-optineurin mutation.
OLs and RGCs were generated from iPSCs derived two healthy patient indivisuals (WT-iPSCs; 201B6 and 201B7 Cell line) and three patients with OPTNE50K-NTG (E50K-iPSCs). The phenotype of WT-iPSCs OLs and E50K-iPSCs OLs was compared by using Western blotting for LC3B, p62, cleaved caspase-3, myelin basic protein (MBP) and nuclear factor-kappaB (NF-κB). In immunostaining analysis, O4 was used as oligodendrocyte precursor cells (OPCs) marker and MBP as mature OLs marker.
MBP-positive mature OLs were produced from iPSCs. Comparing with WT-iPSCs OLs, E50K-iPSCs OLs showed prompting autophagic flux, increased cleaved caspase-3, enhanced MBP insolubility and decreasing MBP-positive area. Moreover, conditioned medium (CM) collected from E50K-iPSCs OLs culture decreased ATH5/TUJ1-positive cells and neurite outgrowth comparing with those of CM collected from WT-iPSCs OLs
Our study demonstrated that E50K-iPSCs OLs showed promoting autophagic flux, abnormal features of MBP protein and increasing cleaved caspase-3, suggesting that E50K-iPSCs OLs itself might enhance vulnerability. Moreover, although secreted soluble factor from WT-iPSCs OLs showed neuroprotective action for WT and E50K-iPSCs RGCs, E50K-iPSCs OLs showed weaker neuroprotective action than WT-iPSCs OLs.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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