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Updated: Oct 20, 2025
Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells
Published on: December 11, 2020
Oxidative Stress Resistance 1 Gene Therapy Retards Neurodegeneration in the rd1 Mutant Mouse Model of Retinopathy
Bhubanananda Sahu1,2, Laura Moreno Leon1,2, Wei Zhang1,2
1Department of Ophthalmology and Visual Science, University of Massachusetts Medical School, Worcester, Massachusetts, United States.
Purpose:
Oxidative stress is a major factor underlying many neurodegenerative diseases. However, antioxidant therapy has had mixed results, possibly because of its indiscriminate activity. The purpose of our study was to determine if the human OXR1 (hOXR1) antioxidant regulatory gene could protect neurons from oxidative stress and delay photoreceptor cell death.
Methods:
The cone-like 661W cell line was transfected to stably express the hOXR1 gene. Oxidative stress was induced by the addition of hydrogen peroxide (H2O2). Intracellular levels of reactive oxygen species (ROS), caspase cleavage, and cellular resistance to oxidative stress were determined and compared between the control and hOXR1 cells. For in vivo analysis, AAV8-hOXR1 was injected subretinally into the rd1 mouse model of retinal degeneration. Functional and structural integrity of the photoreceptors were assessed using electroretinography (ERG), histology, and immunofluorescence analysis.
Results:
Expression of hOXR1 increased cellular resistance and reduced ROS levels and caspase cleavage in the 661W cell line after H2O2-induced oxidative stress. Subretinal injection of AAV8-hOXR1 in the rd1 mice improved their photoreceptor light response, expression and localization of photoreceptor-specific proteins, and delayed retinal degeneration.
Conclusions:
Our results suggest that OXR1 is a potential therapy candidate for retinal degeneration. Because OXR1 targets oxidative stress, a common feature of many retinal degenerative diseases, it should be of therapeutic value to multiple retinal degenerative diseases.
Insights
The human OXR1 gene protects neurons from oxidative stress, reducing cell death in models of neurodegenerative diseases. This antioxidant gene shows promise for treating various retinal degenerative conditions.
Area of Science:
- Neuroscience
- Genetics
- Ophthalmology
Background:
- Oxidative stress is a key factor in neurodegenerative diseases.
- Current antioxidant therapies show limited efficacy due to indiscriminate action.
- Targeted antioxidant gene therapy may offer a more effective approach.
Purpose of the Study:
- To investigate the neuroprotective potential of the human OXR1 (hOXR1) gene.
- To determine if hOXR1 can shield neurons from oxidative stress and photoreceptor cell death.
- To evaluate hOXR1 as a therapeutic agent for retinal degeneration.
Main Methods:
- Stable transfection of 661W cells with hOXR1 and induction of oxidative stress with H2O2.
- Measurement of intracellular reactive oxygen species (ROS) and caspase cleavage.
- Subretinal injection of AAV8-hOXR1 in the rd1 mouse model for in vivo assessment.
Main Results:
- hOXR1 expression enhanced cellular resistance to oxidative stress and reduced ROS and caspase activity.
- In rd1 mice, AAV8-hOXR1 improved photoreceptor function and structure.
- Retinal degeneration was delayed in mice treated with AAV8-hOXR1.
Conclusions:
- The OXR1 gene demonstrates significant potential as a therapeutic candidate for retinal degeneration.
- Targeting oxidative stress with OXR1 offers a promising strategy for multiple retinal degenerative diseases.
- hOXR1 represents a novel therapeutic avenue for combating neurodegeneration in the retina.
