Restoration of visual function in adult mice with an inherited retinal disease via adenine base editing

Suh, Suk-Hwan; Choi, Elliot H.; Leinonen, Henri; Foik, Andrzej T.; Newby, Gregory A.; Yeh, Wei-Hsi; Dong, Zhiqian; Kiser, Philip D.; Lyon, David C.; Liu, David R.; Palczewski, Krzysztof

doi:10.1038/s41551-020-00632-6

Cited by 122 publications

(114 citation statements)

References 53 publications

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“…The SpCas9 guide RNA (gRNA) tethers the ABE to the target site, and the adenine deaminase converts a nearby adenine to hypoxanthine and, ultimately, guanine. Unlike HDR, adenine base editing does not require cells to be proliferating to efficiently introduce mutations and has infrequent unwanted on- and off-target mutagenesis 26 , 27 , offering a potentially safer, more efficient correction of the MPS-IH G→A mutation.…”

Section: Introductionmentioning

confidence: 99%

In utero adenine base editing corrects multi-organ pathology in a lethal lysosomal storage disease

et al. 2021

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In utero base editing has the potential to correct disease-causing mutations before the onset of pathology. Mucopolysaccharidosis type I (MPS-IH, Hurler syndrome) is a lysosomal storage disease (LSD) affecting multiple organs, often leading to early postnatal cardiopulmonary demise. We assessed in utero adeno-associated virus serotype 9 (AAV9) delivery of an adenine base editor (ABE) targeting the Idua G→A (W392X) mutation in the MPS-IH mouse, corresponding to the common IDUA G→A (W402X) mutation in MPS-IH patients. Here we show efficient long-term W392X correction in hepatocytes and cardiomyocytes and low-level editing in the brain. In utero editing was associated with improved survival and amelioration of metabolic, musculoskeletal, and cardiac disease. This proof-of-concept study demonstrates the possibility of efficiently performing therapeutic base editing in multiple organs before birth via a clinically relevant delivery mechanism, highlighting the potential of this approach for MPS-IH and other genetic diseases.

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Section: Introductionmentioning

confidence: 99%

In utero adenine base editing corrects multi-organ pathology in a lethal lysosomal storage disease

et al. 2021

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“…A recent study showed efficient ex vivo editing with high-dose (40 μM) treatment with CBE ( 13 ); in this approach, it was estimated that about 160 μg of CBE protein was used to treat 50,000 hematopoietic stem and progenitor cells. To date, however, the use of RNPs for ABE delivery for biomedical applications has been limited, although delivery via mRNA ( 31 ), plasmid ( 43 ), and viral vector ( 9 , 10 , 44 ) has demonstrated the therapeutic potential of ABE, including a recent study showing successful gene correction of the rd12 mutation and restoration of retinal function in rd12 mice using ABE-encoding lentiviral vector ( 45 ). To the best of our knowledge, our use of RNP-mediated ABE delivery for in vivo gene correction in mice is the first biomedical application of ABE RNPs.…”

Section: Discussionmentioning

confidence: 99%

High-purity production and precise editing of DNA base editing ribonucleoproteins

Jang

Lee

et al. 2021

Sci. Adv.

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Ribonucleoprotein (RNP) complex–mediated base editing is expected to be greatly beneficial because of its reduced off-target effects compared to plasmid- or viral vector–mediated gene editing, especially in therapeutic applications. However, production of recombinant cytosine base editors (CBEs) or adenine base editors (ABEs) with ample yield and high purity in bacterial systems is challenging. Here, we obtained highly purified CBE/ABE proteins from a human cell expression system and showed that CBE/ABE RNPs exhibited different editing patterns (i.e., less conversion ratio of multiple bases to single base) compared to plasmid-encoded CBE/ABE, mainly because of the limited life span of RNPs in cells. Furthermore, we found that off-target effects in both DNA and RNA were greatly reduced for ABE RNPs compared to plasmid-encoded ABE. We ultimately applied NG PAM–targetable ABE RNPs to in vivo gene correction in retinal degeneration 12 (rd12) model mice.

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“…Two studies have already reported the use of base-editing strategies in the retina of mice (Levy et al, 2020;Suh et al, 2021). In the first study, Levy et al reported the use of a splitintein base editor-dual AAV strategy in mice expressing tdTomato only in rod photoreceptor cells (Levy et al, 2020).…”

Section: Precise Correction Of Blindness and Deafness Causal Mutationsmentioning

confidence: 99%

“…Robust base editing was reported in transduced rod photoreceptors suggesting that this delivery system could achieved therapeutic editing efficiencies in vivo. In the second study, a lentivirus expressing the ABE protein and its gRNA was delivered to the RPE of LCA mice model by subretinal injection, leading to ∼16% of correction of a de novo nonsense mutation in the RPE65 gene and the restoration of Rpe65 expression (Suh et al, 2021). To assess the rescue of visual function, retinal cell activity was measured using scotopic electroretinography (ERG), the recovery of 44 and 65% of a-and b-wave amplitudes were observed compared to WT control responses.…”

Section: Precise Correction Of Blindness and Deafness Causal Mutationsmentioning

confidence: 99%

Progress in Gene Editing Tools and Their Potential for Correcting Mutations Underlying Hearing and Vision Loss

Botto

Dalkara²,

El-Amraoui³

2021

Front. Genome Ed.

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Blindness and deafness are the most frequent sensory disorders in humans. Whatever their cause — genetic, environmental, or due to toxic agents, or aging — the deterioration of these senses is often linked to irreversible damage to the light-sensing photoreceptor cells (blindness) and/or the mechanosensitive hair cells (deafness). Efforts are increasingly focused on preventing disease progression by correcting or replacing the blindness and deafness-causal pathogenic alleles. In recent years, gene replacement therapies for rare monogenic disorders of the retina have given positive results, leading to the marketing of the first gene therapy product for a form of childhood hereditary blindness. Promising results, with a partial restoration of auditory function, have also been reported in preclinical models of human deafness. Silencing approaches, including antisense oligonucleotides, adeno-associated virus (AAV)–mediated microRNA delivery, and genome-editing approaches have also been applied to various genetic forms of blindness and deafness The discovery of new DNA- and RNA-based CRISPR/Cas nucleases, and the new generations of base, prime, and RNA editors offers new possibilities for directly repairing point mutations and therapeutically restoring gene function. Thanks to easy access and immune-privilege status of self-contained compartments, the eye and the ear continue to be at the forefront of developing therapies for genetic diseases. Here, we review the ongoing applications and achievements of this new class of emerging therapeutics in the sensory organs of vision and hearing, highlighting the challenges ahead and the solutions to be overcome for their successful therapeutic application in vivo.

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Restoration of visual function in adult mice with an inherited retinal disease via adenine base editing

Cited by 122 publications

References 53 publications

In utero adenine base editing corrects multi-organ pathology in a lethal lysosomal storage disease

In utero adenine base editing corrects multi-organ pathology in a lethal lysosomal storage disease

High-purity production and precise editing of DNA base editing ribonucleoproteins

Progress in Gene Editing Tools and Their Potential for Correcting Mutations Underlying Hearing and Vision Loss

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