학술논문
Shuttle peptide delivers base editor RNPs to rhesus monkey airway epithelial cells in vivo
Document Type
article
Author
Katarina Kulhankova; Soumba Traore; Xue Cheng; Hadrien Benk-Fortin; Stéphanie Hallée; Mario Harvey; Joannie Roberge; Frédéric Couture; Sajeev Kohli; Thomas J. Gross; David K. Meyerholz; Garrett R. Rettig; Bernice Thommandru; Gavin Kurgan; Christine Wohlford-Lenane; Dennis J. Hartigan-O’Connor; Bradley P. Yates; Gregory A. Newby; David R. Liu; Alice F. Tarantal; David Guay; Paul B. McCray
Source
Nature Communications, Vol 14, Iss 1, Pp 1-16 (2023)
Subject
Language
English
ISSN
2041-1723
Abstract
Abstract Gene editing strategies for cystic fibrosis are challenged by the complex barrier properties of airway epithelia. We previously reported that the amphiphilic S10 shuttle peptide non-covalently combined with CRISPR-associated (Cas) ribonucleoprotein (RNP) enabled editing of human and mouse airway epithelial cells. Here, we derive the S315 peptide as an improvement over S10 in delivering base editor RNP. Following intratracheal aerosol delivery of Cy5-labeled peptide in rhesus macaques, we confirm delivery throughout the respiratory tract. Subsequently, we target CCR5 with co-administration of ABE8e-Cas9 RNP and S315. We achieve editing efficiencies of up-to 5.3% in rhesus airway epithelia. Moreover, we document persistence of edited epithelia for up to 12 months in mice. Finally, delivery of ABE8e-Cas9 targeting the CFTR R553X mutation restores anion channel function in cultured human airway epithelia. These results demonstrate the therapeutic potential of base editor delivery with S315 to functionally correct the CFTR R553X mutation in respiratory epithelia.