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CRISPR genome surgery in a novel humanized model for autosomal dominant retinitis pigmentosa.

ABSTRACT: Mutations in rhodopsin (RHO) are the most common causes of autosomal dominant retinitis pigmentosa (adRP), accounting for 20% to 30% of all cases worldwide. However, the high degree of genetic heterogeneity makes development of effective therapies cumbersome. To provide a universal solution to RHO-related adRP, we devised a CRISPR-based, mutation-independent gene ablation and replacement (AR) compound therapy carried by a dual AAV2/8 system. Moreover, we developed a novel hRHO^C110R/hRHO^WT humanized mouse model to assess the AR treatment in vivo. Results show that this humanized RHO mouse model exhibits progressive rod-cone degeneration that phenocopies hRHO^C110R/hRHO^WT patients. In vivo transduction of AR AAV8 dual vectors remarkably ablates endogenous RHO expression and overexpresses exogenous WT hRHO. Furthermore, the administration of AR during adulthood significantly hampers photoreceptor degeneration both histologically and functionally for at least 6 months compared with sole gene replacement or surgical trauma control. This study demonstrates the effectiveness of AR treatment of adRP in the human genomic context while revealing the feasibility of its application for other autosomal dominant disorders.

SUBMITTER: Wu WH

PROVIDER: S-EPMC9077379 | biostudies-literature | 2022 Apr

REPOSITORIES: biostudies-literature

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CRISPR genome surgery in a novel humanized model for autosomal dominant retinitis pigmentosa.

Wu Wen-Hsuan WH Tsai Yi-Ting YT Huang I-Wen IW Cheng Chia-Hua CH Hsu Chun-Wei CW Cui Xuan X Ryu Joseph J Quinn Peter M J PMJ Caruso Salvatore Marco SM Lin Chyuang-Sheng CS Tsang Stephen H SH

Molecular therapy : the journal of the American Society of Gene Therapy 20220210 4

Mutations in rhodopsin (RHO) are the most common causes of autosomal dominant retinitis pigmentosa (adRP), accounting for 20% to 30% of all cases worldwide. However, the high degree of genetic heterogeneity makes development of effective therapies cumbersome. To provide a universal solution to RHO-related adRP, we devised a CRISPR-based, mutation-independent gene ablation and replacement (AR) compound therapy carried by a dual AAV2/8 system. Moreover, we developed a novel hRHO<sup>C110R</sup>/hR ...[more]

PMID: 35150888

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Project description:PurposeTo develop a universal gene therapy to overcome the genetic heterogeneity in retinitis pigmentosa (RP) resulting from mutations in rhodopsin (RHO).DesignExperimental study for a combination gene therapy that uses both gene ablation and gene replacement.ParticipantsThis study included 2 kinds of human RHO mutation knock-in mouse models: RhoP23H and RhoD190N. In total, 23 RhoP23H/P23H, 43 RhoP23H/+, and 31 RhoD190N/+ mice were used for analysis.MethodsThis study involved gene therapy using dual adeno-associated viruses (AAVs) that (1) destroy expression of the endogenous Rho gene in a mutation-independent manner via an improved clustered regularly interspaced short palindromic repeats-based gene deletion and (2) enable expression of wild-type protein via exogenous cDNA.Main outcome measuresElectroretinographic and histologic analysis.ResultsThe thickness of the outer nuclear layer (ONL) after the subretinal injection of combination ablate-and-replace gene therapy was approximately 17% to 36% more than the ONL thickness resulting from gene replacement-only therapy at 3 months after AAV injection. Furthermore, electroretinography results demonstrated that the a and b waves of both RhoP23H and RhoD190N disease models were preserved more significantly using ablate-and-replace gene therapy (P < 0.001), but not by gene replacement monotherapy.ConclusionsAs a proof of concept, our results suggest that the ablate-and-replace strategy can ameliorate disease progression as measured by photoreceptor structure and function for both of the human mutation knock-in models. These results demonstrate the potency of the ablate-and-replace strategy to treat RP caused by different Rho mutations. Furthermore, because ablate-and-replace treatment is mutation independent, this strategy may be used to treat a wide array of dominant diseases in ophthalmology and other fields. Clinical trials using ablate-and-replace gene therapy would allow researchers to determine if this strategy provides any benefits for patients with diseases of interest.

Dataset Information

CRISPR genome surgery in a novel humanized model for autosomal dominant retinitis pigmentosa.

Publications

CRISPR genome surgery in a novel humanized model for autosomal dominant retinitis pigmentosa.

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