Functional correction of CFTR mutations in human airway epithelial cells using adenine base editors

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

doi:10.1093/nar/gkab788

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Functional correction of CFTR mutations in human airway epithelial cells using adenine base editors

Journal article

Open access

Peer reviewed

Functional correction of CFTR mutations in human airway epithelial cells using adenine base editors

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

Nucleic acids research, Vol.49(18), pp.10558-10572

10/11/2021

DOI: 10.1093/nar/gkab788

PMCID: PMC8501978

PMID: 34520545

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Abstract

Mutations in the CFTR gene that lead to premature stop codons or splicing defects cause cystic fibrosis (CF) and are not amenable to treatment by small-molecule modulators. Here, we investigate the use of adenine base editor (ABE) ribonucleoproteins (RNPs) that convert A•T to G•C base pairs as a therapeutic strategy for three CF-causing mutations. Using ABE RNPs, we corrected in human airway epithelial cells premature stop codon mutations (R553X and W1282X) and a splice-site mutation (3849 + 10 kb C > T). Following ABE delivery, DNA sequencing revealed correction of these pathogenic mutations at efficiencies that reached 38-82% with minimal bystander edits or indels. This range of editing was sufficient to attain functional correction of CFTR-dependent anion channel activity in primary epithelial cells from CF patients and in a CF patient-derived cell line. These results demonstrate the utility of base editor RNPs to repair CFTR mutations that are not currently treatable with approved therapeutics.

Mutation

Adenine

Cell Line

Cells, Cultured

Cystic Fibrosis - genetics

Cystic Fibrosis Transmembrane Conductance Regulator - genetics

Cystic Fibrosis Transmembrane Conductance Regulator - metabolism

Gene Editing

Humans

Respiratory Mucosa - metabolism

Ribonucleoproteins

Details

Title: Subtitle: Functional correction of CFTR mutations in human airway epithelial cells using adenine base editors
Creators: Sateesh Krishnamurthy - University of Iowa
Soumba Traore - University of Iowa
Ashley L Cooney - University of Iowa
Christian M Brommel - University of Iowa
Katarina Kulhankova - University of Iowa
Patrick L Sinn - University of Iowa
Gregory A Newby - Harvard University
David R Liu - Broad Institute
Paul B McCray - University of Iowa
Resource Type: Journal article
Publication Details: Nucleic acids research, Vol.49(18), pp.10558-10572
DOI: 10.1093/nar/gkab788
PMID: 34520545
PMCID: PMC8501978
NLM abbreviation: Nucleic Acids Res
ISSN: 0305-1048
eISSN: 1362-4962
Grant note: R35 GM118062 / NIGMS NIH HHS P01 HL152960 / NHLBI NIH HHS U01 AI142756 / NIAID NIH HHS UG3 AI150551 / NIAID NIH HHS U24 HG010423 / NHGRI NIH HHS Howard Hughes Medical Institute UG3 HL147366 / NIH HHS P30 DK054759 / NIDDK NIH HHS UG3 TR002636 / NCATS NIH HHS R01 HL133089 / NHLBI NIH HHS
Language: English
Date published: 10/11/2021
Academic Unit: Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Internal Medicine
Record Identifier: 9984297321402771

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