Shuttle peptide delivers base editor RNPs to rhesus monkey airway epithelial cells in vivo

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

doi:10.1038/s41467-023-43904-w

Shuttle peptide delivers base editor RNPs to rhesus monkey airway epithelial cells in vivo

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. TarantalDavid Guay, Paul B. McCray

School of Medicine

Research output: Contribution to journal › Article › peer-review

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.

Original language	English (US)
Article number	8051
Journal	Nature communications
Volume	14
Issue number	1
DOIs	https://doi.org/10.1038/s41467-023-43904-w
State	Published - Dec 2023

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

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Kulhankova, K., Traore, S., Cheng, X., Benk-Fortin, H., Hallée, S., Harvey, M., Roberge, J., Couture, F., Kohli, S., Gross, T. J., Meyerholz, D. K., Rettig, G. R., Thommandru, B., Kurgan, G., Wohlford-Lenane, C., Hartigan-O’Connor, D. J., Yates, B. P., Newby, G. A., Liu, D. R., ... McCray, P. B. (2023). Shuttle peptide delivers base editor RNPs to rhesus monkey airway epithelial cells in vivo. Nature communications, 14(1), Article 8051. https://doi.org/10.1038/s41467-023-43904-w

Kulhankova, K, Traore, S, Cheng, X, Benk-Fortin, H, Hallée, S, Harvey, M, Roberge, J, Couture, F, Kohli, S, Gross, TJ, Meyerholz, DK, Rettig, GR, Thommandru, B, Kurgan, G, Wohlford-Lenane, C, Hartigan-O’Connor, DJ, Yates, BP , Newby, GA, Liu, DR, Tarantal, AF, Guay, D & McCray, PB 2023, 'Shuttle peptide delivers base editor RNPs to rhesus monkey airway epithelial cells in vivo', Nature communications, vol. 14, no. 1, 8051. https://doi.org/10.1038/s41467-023-43904-w

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title = "Shuttle peptide delivers base editor RNPs to rhesus monkey airway epithelial cells in vivo",

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.",

author = "Katarina Kulhankova and Soumba Traore and Xue Cheng and Hadrien Benk-Fortin and St{\'e}phanie Hall{\'e}e and Mario Harvey and Joannie Roberge and Fr{\'e}d{\'e}ric Couture and Sajeev Kohli and Gross, {Thomas J.} and Meyerholz, {David K.} and Rettig, {Garrett R.} and Bernice Thommandru and Gavin Kurgan and Christine Wohlford-Lenane and Hartigan-O{\textquoteright}Connor, {Dennis J.} and Yates, {Bradley P.} and Newby, {Gregory A.} and Liu, {David R.} and Tarantal, {Alice F.} and David Guay and McCray, {Paul B.}",

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doi = "10.1038/s41467-023-43904-w",

language = "English (US)",

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AU - Kulhankova, Katarina

AU - Traore, Soumba

AU - Cheng, Xue

AU - Benk-Fortin, Hadrien

AU - Hallée, Stéphanie

AU - Harvey, Mario

AU - Roberge, Joannie

AU - Couture, Frédéric

AU - Kohli, Sajeev

AU - Gross, Thomas J.

AU - Meyerholz, David K.

AU - Rettig, Garrett R.

AU - Thommandru, Bernice

AU - Kurgan, Gavin

AU - Wohlford-Lenane, Christine

AU - Hartigan-O’Connor, Dennis J.

AU - Yates, Bradley P.

AU - Newby, Gregory A.

AU - Liu, David R.

AU - Tarantal, Alice F.

AU - Guay, David

AU - McCray, Paul B.

PY - 2023/12

Y1 - 2023/12

N2 - 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.

AB - 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.

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Shuttle peptide delivers base editor RNPs to rhesus monkey airway epithelial cells in vivo

Abstract

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