June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Epigenetic reprogramming to rejuvenate retinal ganglion cells and sustainably reverse glaucoma-induced vision loss
Author Affiliations & Notes
  • Yuancheng Ryan Lu
    Whitehead Institute, Cambridge, Massachusetts, United States
  • Margarete Karg
    Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, Massachusetts, United States
    Harvard Medical School, Boston, Massachusetts, United States
  • Emma M. Hoffmann
    Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, Massachusetts, United States
  • James C. Cameron
    Whitehead Institute, Cambridge, Massachusetts, United States
  • May Moorefield
    Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, Massachusetts, United States
  • Maleeka Shrestha
    Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, Massachusetts, United States
  • Yinjie Guo
    Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, Massachusetts, United States
  • Joan Mannick
    Life Biosciences, Boston, Massachusetts, United States
  • Jennifer Cermak
    Life Biosciences, Boston, Massachusetts, United States
  • Michel Wathier
    Life Biosciences, Boston, Massachusetts, United States
  • Meredith S Gregory-Ksander
    Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, Massachusetts, United States
    Harvard Medical School, Boston, Massachusetts, United States
  • David Sinclair
    Harvard Medical School, Boston, Massachusetts, United States
  • Bruce Ksander
    Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, Massachusetts, United States
    Harvard Medical School, Boston, Massachusetts, United States
  • Footnotes
    Commercial Relationships   Yuancheng Lu Life Biosciences, Code I (Personal Financial Interest), Life Biosciences, Code P (Patent); Margarete Karg None; Emma Hoffmann None; James Cameron None; May Moorefield None; Maleeka Shrestha None; Yinjie Guo None; Joan Mannick Life Biosciences, Code E (Employment), Tornado Therapeutics , Code O (Owner); Jennifer Cermak Life Biosciences, Code E (Employment); Michel Wathier Life Biosciences, Code E (Employment); Meredith Gregory-Ksander None; David Sinclair Life Biosciences, Code I (Personal Financial Interest), Life Biosciences, Code P (Patent); Bruce Ksander Life Biosciences, Code F (Financial Support)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 2841. doi:
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      Yuancheng Ryan Lu, Margarete Karg, Emma M. Hoffmann, James C. Cameron, May Moorefield, Maleeka Shrestha, Yinjie Guo, Joan Mannick, Jennifer Cermak, Michel Wathier, Meredith S Gregory-Ksander, David Sinclair, Bruce Ksander; Epigenetic reprogramming to rejuvenate retinal ganglion cells and sustainably reverse glaucoma-induced vision loss. Invest. Ophthalmol. Vis. Sci. 2023;64(8):2841.

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

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Abstract

Purpose : We previously reported that in vivo epigenetic reprogramming by AAV2-tTA; TRE-OSK (Oct4, Sox2, and Klf4) reversed the age of retinal ganglion cells (RGCs) in old mice based on transcriptome and DNA methylome. This led to a significant improvement in RGC function and vision. In addition, OSK reprogramming restored the axon regeneration ability of RGCs and improved the vision in mice with microbead-induced glaucoma (Lu, et al, Nature, 2020). In this study, we performed the first long-term functional tracking study for in vivo reprogramming and addressed the following questions: in the context of glaucoma, what is the maximum duration of the reprogramming effect; can it be achieved using a Doxycycline (Dox)-inducible dual AAV system; and is long term in vivo reprogramming effective and safe?

Methods : Glaucoma was induced in C57BL/6 mice by an intracameral injection of magnetic microbeads. Four weeks post-microbead injection, doxycycline (Dox) inducible OSK vectors containing a 1:1 mixture of AAV2-TRE-OSK, together with either AAV2-tTA (Dox-OFF) or AAV2-rtTA (Dox-ON) were injected intravitreally (1μl). Dox (2mg/mL) was delivered via drinking water to induce AAV2-rtTA and TRE-OSK expression. The effect of long-term continuous expression of OSK was monitored in the AAV2-tTA; TRE-OSK group. Non-glaucomatous healthy mice were included to record the damage-free baseline during the one-year span. OMR was measured monthly for a year.

Results : In the AAV2-tTA; TRE-OSK mice (continuously on), vision lost from glaucoma was completely restored to healthy levels after 2 months of treatment and remained close to healthy levels for 11 months. In the AAV2-rtTA; TRE-OSK group, while the vision of DOX untreated mice continued to decline due to glaucoma, the mice treated with Dox and OSK also completely restored vision to healthy levels by 8 weeks, suggesting OSK beneficial effect can be achieved using a Dox-inducible dual AAV system. When Dox was withdrawn, the improved vision remained significantly better than untreated baselines. There was no adverse effect observed in mice expressing OSK over the entire course.

Conclusions : This first long-term functional tracking study for in vivo reprogramming indicates that epigenetic reprogramming of RGCs is safe, and induced sustained recovery of visual function that was lost due to elevated pressure-induced glaucoma.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

 

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