High-Efficiency, Selection-free Gene Repair in Airway Stem Cells from Cystic Fibrosis Patients Rescues CFTR Function in Differentiated Epithelia

Vaidyanathan, Sriram; Salahudeen, Ameen A.; Sellers, Zachary M.; Bravo, Dawn T.; Choi, Stacey S.; Batish, Arpit; Le, Wei; Baik, Ron; O, Sean de la; Kaushik, Milan P.; Galper, Noah; Lee, Ciaran M.; Teran, Christopher A.; Yoo, Jessica H.; Bao, Gang; Chang, Eugene H.; Patel, Zara M.; Hwang, Peter H.; Wine, Jeffrey J.; Milla, Carlos; Desai, Tushar J.; Nayak, Jayakar V.; Kuo, Calvin J.; Porteus, Matthew H.

doi:10.1016/j.stem.2019.11.002

Cited by 107 publications

(97 citation statements)

References 52 publications

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“…Without selection, from 30% to 40% of the mutated alleles were repaired in primary airway stem cells showing functional recovery of the CFTR channel. In this study the edited cells were embedded in FDA-approved porcine epithelial small intestinal submucosal membrane (pSIS) obtaining large number of CFTR expressing cells, potentially usable for transplantation in the upper-airway of CF patients [56]. These are encouraging data on the preserved nature of stem cells after genome editing in spite of p53 activation reported to occur in hematopoietic stem cells after DSB generated by programmable nucleases [57].…”

Section: Gene Correction By Homology Directed Repairmentioning

confidence: 98%

Gene Therapy for Cystic Fibrosis: Progress and Challenges of Genome Editing

Maule

Arosio

Cereseto

2020

IJMS

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Since the early days of its conceptualization and application, human gene transfer held the promise of a permanent solution to genetic diseases including cystic fibrosis (CF). This field went through alternated periods of enthusiasm and distrust. The development of refined technologies allowing site specific modification with programmable nucleases highly revived the gene therapy field. CRISPR nucleases and derived technologies tremendously facilitate genome manipulation offering diversified strategies to reverse mutations. Here we discuss the advancement of gene therapy, from therapeutic nucleic acids to genome editing techniques, designed to reverse genetic defects in CF. We provide a roadmap through technologies and strategies tailored to correct different types of mutations in the cystic fibrosis transmembrane regulator (CFTR) gene, and their applications for the development of experimental models valuable for the advancement of CF therapies.

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Section: Gene Correction By Homology Directed Repairmentioning

confidence: 98%

Gene Therapy for Cystic Fibrosis: Progress and Challenges of Genome Editing

Maule

Arosio

Cereseto

2020

IJMS

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“…From a more clinically oriented perspective, tissue replacement using gene-corrected organoids holds great promise for restoring vital tissue functions that are impaired by pathogenic genetic variants. Correction of defective CFTR in airway organoids derived from patients with cystic fibrosis 161 and their transplantation may alleviate respiratory complications. Patients with congenital metabolic diseases that result from an insufficient amount or function of vital enzymes may benefit from transplantation of gene-corrected liver organoids.…”

Section: Perspectives and Conclusionmentioning

confidence: 99%

Somatic cell-derived organoids as prototypes of human epithelial tissues and diseases

2020

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“…They achieved 30-50% editing efficiencies that restored CFTR function equivalent to 20-50% of wild-type controls. 62 Finally, scientists used helper-dependent adenoviruses to deliver Cas9, guide and template DNA to iPSCs, significantly improving gene editing efficiencies. While the majority of recombinants were able to integrate the donor DNA into one of the mutated alleles, the second allele had a high rate of indels at the Cas9 cleavage site, sometimes up to 8 kb deletions.…”

Section: Rna Therapies Can Be Used To Replace or Edit Cftrmentioning

confidence: 99%

Treating Cystic Fibrosis with mRNA and CRISPR

et al. 2020

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Less than 20% of the protein coding genome is thought to be targetable using small molecules. mRNA therapies are not limited in the same way since in theory, they can silence or edit any gene by encoding CRISPR nucleases, or alternatively, produce any missing protein. Yet not all mRNA therapies are equally likely to succeed. Over the past several years, an increasing number of clinical trials with siRNA-and antisense oligonucleotide-based drugs have revealed three key concepts that will likely extend to mRNA therapies delivered by nonviral systems. First, scientists have come to understand that some genes make better targets for RNA therapies than others. Second, scientists have learned that the type and position of chemical modifications made to an RNA drug can alter its therapeutic window, toxicity, and bioavailability. Third, scientists have found that safe and targeted drug delivery vehicles are required to ferry mRNA therapies into diseased cells. In this study, we apply these learnings to cystic fibrosis (CF). We also describe lessons learned from a subset of CF gene therapies that have already been tested in patients. Finally, we highlight the scientific advances that are still required for nonviral mRNA-or CRISPR-based drugs to treat CF successfully in patients.

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High-Efficiency, Selection-free Gene Repair in Airway Stem Cells from Cystic Fibrosis Patients Rescues CFTR Function in Differentiated Epithelia

Cited by 107 publications

References 52 publications

Gene Therapy for Cystic Fibrosis: Progress and Challenges of Genome Editing

Gene Therapy for Cystic Fibrosis: Progress and Challenges of Genome Editing

Somatic cell-derived organoids as prototypes of human epithelial tissues and diseases

Treating Cystic Fibrosis with mRNA and CRISPR

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