Functional correction of<i>CFTR</i>mutations in human airway epithelial cells using adenine base editors

Krishnamurthy, Sateesh; Traore, Soumba; Cooney, Ashley L.; Brommel, Christian M; Kulhánková, Katarina; Sinn, Patrick L.; Newby, Gregory A.; Liu, David R.; McCray, Paul B.

doi:10.1093/nar/gkab788

Cited by 27 publications

(23 citation statements)

References 95 publications

(105 reference statements)

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“…Base or prime editing, both CRISPR-derived gene editing technologies (reviewed in [ 267 , 357 ]), allow to make precise, targeted edits in the genome, and are more likely able to correct these mutations. In that regard, successful adenine base editing has already been already reported for correcting nonsense mutations in CFTR [ 354 , 379 ]. In addition, frameshift mutations caused by small indels might be rescued by base or prime editing approaches in the future.…”

Section: Towards the Future: Personalizing Therapies For Pwcfmentioning

confidence: 99%

One Size Does Not Fit All: The Past, Present and Future of Cystic Fibrosis Causal Therapies

Ensinck

Carlon

2022

Cells

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Cystic fibrosis (CF) is the most common monogenic disorder, caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. Over the last 30 years, tremendous progress has been made in understanding the molecular basis of CF and the development of treatments that target the underlying defects in CF. Currently, a highly effective CFTR modulator treatment (Kalydeco™/Trikafta™) is available for 90% of people with CF. In this review, we will give an extensive overview of past and ongoing efforts in the development of therapies targeting the molecular defects in CF. We will discuss strategies targeting the CFTR protein (i.e., CFTR modulators such as correctors and potentiators), its cellular environment (i.e., proteostasis modulation, stabilization at the plasma membrane), the CFTR mRNA (i.e., amplifiers, nonsense mediated mRNA decay suppressors, translational readthrough inducing drugs) or the CFTR gene (gene therapies). Finally, we will focus on how these efforts can be applied to the 15% of people with CF for whom no causal therapy is available yet.

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Section: Towards the Future: Personalizing Therapies For Pwcfmentioning

confidence: 99%

One Size Does Not Fit All: The Past, Present and Future of Cystic Fibrosis Causal Therapies

Ensinck

Carlon

2022

Cells

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“…3D). Five days after the enrichment of transfected cells (EGFP + sorted cells), the BE efficiency was estimated by DNA-sequencing as previously described 6,32 . CRISPResso2 analyses indicated that A907 was converted into G907 (reference nucleotide) in ∼52% of reads from the maternal allele as expected with this BE approach (FIG.…”

Section: Resultsmentioning

confidence: 99%

“…In total, 978 candidate sites have been predicted, including only 63 exonic sites (Table S3). Since it remains difficult to evaluate all these potential sites, we ranked them according to the Cutting Frequency Determination (CFD) score as previously published 6 . We focused on the top-10 CFD scores (only one is located in an exon ( IDH1 gene, Table S3)) and off-target effects were investigated by deep-sequencing (amplicon-seq).…”

Section: Resultsmentioning

confidence: 99%

“…Nowadays, gene editing approaches such as base and prime editing (BE and PE) enable specific restoration of a functional gene by correcting these deleterious variants 3 . BE has been successfully used to cure different diseases in different models and species [4][5][6][7][8] . To date, the most advanced application in humans is to cure sickle cell anemia via ex vivo targeting of haematopoietic stem and progenitor cells (HSPCs) from patients 5 .…”

Section: Introductionmentioning

confidence: 99%

“…France. 6 : Laboratoire de Génétique Moléculaire et Génomique, Centre Hospitalier Universitaire de Rennes, Rennes. 7 : Univ Rennes, CNRS, IGDR (Institut de Génétique et Développement de Rennes), UMR 6290, ERL U1305, Rennes, France.…”

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Gene editing is suitable to treat GM1 Gangliosidosis: a proof-of-concept study

Leclerc

Goujon

Jaillard

et al. 2022

Preprint

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Ganglioside-monosialic acid (GM1) gangliosidosis, a rare autosomal recessive disorder, is frequently caused by deleterious single nucleotide variants (SNVs) in GLB1 gene. These variants result in reduced beta-galactosidase (β-gal) activity, leading to neurodegeneration associated with premature death. Currently, no effective therapy for GM1 gangliosidosis is available. Three ongoing clinical trials aim to deliver a functional copy of the GLB1 gene to stop disease progression. Here, we show that 41% of GLB1 pathogenic SNVs might be cured by adenine base editors (ABEs). Our results demonstrate that ABE efficiently corrects the pathogenic allele in patient-derived fibroblasts, restoring a therapeutic level of β-gal activity. Unbiased off-target DNA analysis did not detect off-target editing activity in treated patient cells except a bystander edit without consequences on β-gal activity. Altogether our results suggest that gene editing is an alternative strategy to cure GM1 gangliosidosis, by correcting the root cause of disease and avoiding repetitive adeno-associated virus injections.

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Optimization of ionizable lipids for aerosolizable mRNA lipid nanoparticles

Lewis,

Soto,

Maier

et al. 2023

Bioengineering & Transla Med

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Although mRNA lipid nanoparticles (LNPs) are highly effective as vaccines, their efficacy for pulmonary delivery has not yet fully been established. A major barrier to this therapeutic goal is their instability during aerosolization for local delivery. This imparts a shear force that degrades the mRNA cargo and therefore reduces cell transfection. In addition to remaining stable upon aerosolization, mRNA LNPs must also possess the aerodynamic properties to achieve deposition in clinically relevant areas of the lungs. We addressed these challenges by formulating mRNA LNPs with SM‐102, the clinically approved ionizable lipid in the Spikevax COVID‐19 vaccine. Our lead candidate, B‐1, had the highest mRNA expression in both a physiologically relevant air–liquid interface (ALI) human lung cell model and in healthy mice lungs upon aerosolization. Further, B‐1 showed selective transfection in vivo of lung epithelial cells compared to immune cells and endothelial cells. These results show that the formulation can target therapeutically relevant cells in pulmonary diseases such as cystic fibrosis. Morphological studies of B‐1 revealed differences in the surface structure compared to LNPs with lower transfection efficiency. Importantly, the formulation maintained critical aerodynamic properties in simulated human airways upon next generation impaction. Finally, structure–function analysis of SM‐102 revealed that small changes in the number of carbons can improve upon mRNA delivery in ALI human lung cells. Overall, our study expands the application of SM‐102 and its analogs to aerosolized pulmonary delivery and identifies a potent lead candidate for future therapeutically active mRNA therapies.

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Functional correction ofCFTRmutations in human airway epithelial cells using adenine base editors

Cited by 27 publications

References 95 publications

One Size Does Not Fit All: The Past, Present and Future of Cystic Fibrosis Causal Therapies

One Size Does Not Fit All: The Past, Present and Future of Cystic Fibrosis Causal Therapies

Gene editing is suitable to treat GM1 Gangliosidosis: a proof-of-concept study

Optimization of ionizable lipids for aerosolizable mRNA lipid nanoparticles

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