June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
A new mode of gene therapy to subdue the limitations of the allotopic method: an analytic approach
Author Affiliations & Notes
  • sindhu velmurugan
    ophthalmology, University of Miami School of Medicine, Miami, Florida, United States
  • Tsung-Han Chou
    ophthalmology, University of Miami School of Medicine, Miami, Florida, United States
  • Jeremy D. Eastwood
    ophthalmology, University of Miami School of Medicine, Miami, Florida, United States
  • Diego Alba
    ophthalmology, University of Miami School of Medicine, Miami, Florida, United States
  • Vittorio Porciatti
    ophthalmology, University of Miami School of Medicine, Miami, Florida, United States
  • John Guy
    ophthalmology, University of Miami School of Medicine, Miami, Florida, United States
  • Hong Yu
    ophthalmology, University of Miami School of Medicine, Miami, Florida, United States
  • Footnotes
    Commercial Relationships   sindhu velmurugan None; Tsung-Han Chou None; Jeremy D. Eastwood None; Diego Alba None; Vittorio Porciatti None; John Guy None; Hong Yu None
  • Footnotes
    Support  R01 EY 027414, R01 EY017141, R24 EY028785, P30 EY014801
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 2830 – A0346. doi:
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      sindhu velmurugan, Tsung-Han Chou, Jeremy D. Eastwood, Diego Alba, Vittorio Porciatti, John Guy, Hong Yu; A new mode of gene therapy to subdue the limitations of the allotopic method: an analytic approach. Invest. Ophthalmol. Vis. Sci. 2022;63(7):2830 – A0346.

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

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Abstract

Purpose : To compare the efficacy of gene therapy using mito-targeted and allotopic expression strategies to treat LHON in mice.

Methods : Adult DBA/1J mice (N=30) were randomly split into three groups and were injected with mitochondria-targeted AAV carrying mutant human ND4 gene (hND4G11778A) to induce LHON. We rescued them at a two-day interval using 1) mito-targeted strategy with MTSAAVs carrying- a) mCherry(n=10), b) wtND4(n=10), naïve (mCherry-mCherry) (n=10) and 2) allotopic strategy (n=20) with MTSAAV carrying a) mCherry (n=10), b) wtND4 (n=10) into both eyes. The rescue of the optic nerve and retina atrophy was evaluated using PERG and TEM.

Results : Our main hypothesis is that the mito-targeted strategy will likely result in a higher delivery efficacy than the allotopic approach making it a quicker and an efficient treatment option for LHON. PERG was performed and mice treated with mito-targeted wildtype hND4 showed a PERG amplitude increase by 34% (p=0.150), 46% (p=0.035), and 83% (p=0.093) respectively at 3, 6, and 12 months after injection, whereas mice treated with allotopic expressed wildtype hND4 increased PERG amplitude by 12% (p=0.594), 21% (p=0.357), 43% (p= 0.012), and 33% (p=0.087).The time course of age-related PERG changes was different between the 2 groups (Interaction age x treatment, P=0.003) (Fig.1A), indicating the efficiency of mito-targeted rescue over allotopic where no significant effect of age (P=0.55) but an overall effect of group on PERG amplitude between the rescued and unrescued mice(P=0.043) was observed (Fig.1B). The significance in amplitude difference (control vs rescue) for mice in mito-targeted strategy (OD, P=0.006; OS, P=0.023) indicates that the rescue efficacy with mito-targeted ND4 compared to allotopic.
TEM revealed that MTND4-rescued mice displayed a shift towards small axons showing that the mito-targeted hND4 efficiently prevented the demise of small axons that are lost in human LHON.

Conclusions : Our data showed that the severe visual loss induced by a mitochondrial disease may be reversed using both mito-targeted and allotopic expressed gene therapy,but mito-targeted therapy likely mediates a quicker and more efficient rescue than the allotopic strategy making it a promising option to treat LHON.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

 

Mean of PERG amplitudes of all mice tested in allotopic (A) group.

Mean of PERG amplitudes of all mice tested in allotopic (A) group.

 

Mean of PERG amplitudes of all mice tested in mito-targeted(B) group.

Mean of PERG amplitudes of all mice tested in mito-targeted(B) group.

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