June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Characterization of Optineurin E50K knock-in mouse model with normal tension glaucoma and evaluation of TBK1 inhibitor Amlexanox
Author Affiliations & Notes
  • Mao Nakayama
    Molecular & Cellular Biol, Nat'l Hosp Org Tokyo Med Ctr, Tokyo, Japan
  • Yuriko Minegishi
    Molecular & Cellular Biol, Nat'l Hosp Org Tokyo Med Ctr, Tokyo, Japan
  • Daisuke Iejima
    Molecular & Cellular Biol, Nat'l Hosp Org Tokyo Med Ctr, Tokyo, Japan
  • Takeshi Iwata
    Molecular & Cellular Biol, Nat'l Hosp Org Tokyo Med Ctr, Tokyo, Japan
  • Footnotes
    Commercial Relationships   Mao Nakayama, None; Yuriko Minegishi, None; Daisuke Iejima, None; Takeshi Iwata, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 5623. doi:
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      Mao Nakayama, Yuriko Minegishi, Daisuke Iejima, Takeshi Iwata; Characterization of Optineurin E50K knock-in mouse model with normal tension glaucoma and evaluation of TBK1 inhibitor Amlexanox. Invest. Ophthalmol. Vis. Sci. 2017;58(8):5623.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Optineurin (OPTN), a component of autophagosome, is responsible for normal tension glaucoma (NTG). In this study, OPTN E50K knock-in mouse was generated by CRISPR/Cas9 and analyzed for abnormality of the retina. Furthermore, we investigated neuroprotective effect of TBK1 inhibitor, Amlexanox, to these E50K knock-in mice.

Methods : Imaging of fundus and retina section of wild-type, homozygous and heterozygous E50K knock-in mice was analyzed. The mouse optic nerve was scanned by optical coherence tomography (OCT, Micron IV, Phoenix Research) and the average thickness of the optic retinal ganglion cells (RGC) layer was analyzed. Immunohistochemistry of E50K knock-in mouse retina was stained with antibodies against anti-βIII tubulin (Chemicon) and anti-SMI32 (Sternberger). Additionally, TBK1 inhibitor Amlexanox was administrated to E50K knock-in mice from 4-week old. After 12 months of administration, we analyzed the average thickness of optic RGC layer to evaluate Amlexanox neuroprotection in E50K knock-in mouse.

Results : The average thickness (57.8μm) of optic RGC layer in E50K knock-in homozygous mice at 6 months shows a significant reduction compared with wild-type mice (75.7μm). β-III tubulin-stained nerve fiber in E50K knock-in homozygous mouse at 12 months shows thinning of the optic nerve fiber compared with wild-type mouse. Furthermore, the optic cupping increased in E50K knock-in homozygous mouse, which mimic NTG patients. SMI32 immunostaining of whole-retina mount also shows RGC loss in E50K knock-in homozygous mouse. The homozygous E50K knock-in mice without Amlexanox treatment exhibited a 15% reduction of the optic RGC layer thickness whereas the treated group at 3%.

Conclusions : Homozygous E50K knock-in mice is the first normal tension glaucoma mouse model, which exhibits the glaucoma-like phenotype by exact same mutation in patient. Amlexanox, which has long been used to treat bronchial asthma and allergic rhinitis as FDA approved drug, rescued the RGC thinning of E50K mouse, indicating as potential drug to treat OPTN E50K glaucoma patients worldwide.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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