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Published on: December 22, 2014
Cone-Specific Promoters for Gene Therapy of Achromatopsia and Other Retinal Diseases
Guo-Jie Ye1, Ewa Budzynski2, Peter Sonnentag2
11 Applied Genetic Technologies Corporation , Alachua, Florida.
Abstract:
Adeno-associated viral (AAV) vectors containing cone-specific promoters have rescued cone photoreceptor function in mouse and dog models of achromatopsia, but cone-specific promoters have not been optimized for use in primates. Using AAV vectors administered by subretinal injection, we evaluated a series of promoters based on the human L-opsin promoter, or a chimeric human cone transducin promoter, for their ability to drive gene expression of green fluorescent protein (GFP) in mice and nonhuman primates. Each of these promoters directed high-level GFP expression in mouse photoreceptors. In primates, subretinal injection of an AAV-GFP vector containing a 1.7-kb L-opsin promoter (PR1.7) achieved strong and specific GFP expression in all cone photoreceptors and was more efficient than a vector containing the 2.1-kb L-opsin promoter that was used in AAV vectors that rescued cone function in mouse and dog models of achromatopsia. A chimeric cone transducin promoter that directed strong GFP expression in mouse and dog cone photoreceptors was unable to drive GFP expression in primate cones. An AAV vector expressing a human CNGB3 gene driven by the PR1.7 promoter rescued cone function in the mouse model of achromatopsia. These results have informed the design of an AAV vector for treatment of patients with achromatopsia.
Insights
Adeno-associated viral vectors show promise for achromatopsia treatment. A novel L-opsin promoter effectively drives gene expression in primate cone photoreceptors, advancing therapy development.
Area of Science:
- Ophthalmology
- Gene Therapy
- Molecular Biology
Background:
- Achromatopsia is an inherited retinal disorder affecting cone photoreceptors.
- Adeno-associated viral (AAV) vectors are explored for gene therapy, but effective cone-specific promoters are needed, especially for primate models.
Purpose of the Study:
- To evaluate and optimize cone-specific promoters for AAV vectors in primate models of achromatopsia.
- To identify a promoter suitable for driving therapeutic gene expression in human cone photoreceptors.
Main Methods:
- Subretinal injection of AAV vectors carrying green fluorescent protein (GFP) reporter gene.
- Evaluation of L-opsin and chimeric cone transducin promoters in mouse and nonhuman primate models.
- Assessment of promoter efficiency and specificity for cone photoreceptor gene expression.
Main Results:
- The 1.7-kb human L-opsin promoter (PR1.7) demonstrated strong, specific GFP expression in primate cone photoreceptors.
- PR1.7 was more efficient than a previously used 2.1-kb L-opsin promoter in primates.
- A chimeric cone transducin promoter failed to drive gene expression in primate cones.
Conclusions:
- The PR1.7 promoter is a highly efficient and specific tool for driving gene expression in primate cone photoreceptors.
- This finding is crucial for developing AAV-based gene therapies for achromatopsia.
- Optimized AAV vectors using PR1.7 can advance treatment strategies for achromatopsia patients.
