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Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia

The delivery of biologic cargoes to airway epithelial cells is challenging due to the formidable barriers imposed by its specialized and differentiated cells. Among cargoes, recombinant proteins offer therapeutic promise but the lack of effective delivery methods limits their development. Here, we a...

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Autores principales: Krishnamurthy, Sateesh, Wohlford-Lenane, Christine, Kandimalla, Suhas, Sartre, Gilles, Meyerholz, David K., Théberge, Vanessa, Hallée, Stéphanie, Duperré, Anne-Marie, Del’Guidice, Thomas, Lepetit-Stoffaes, Jean-Pascal, Barbeau, Xavier, Guay, David, McCray, Paul B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6817825/
https://www.ncbi.nlm.nih.gov/pubmed/31659165
http://dx.doi.org/10.1038/s41467-019-12922-y
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author Krishnamurthy, Sateesh
Wohlford-Lenane, Christine
Kandimalla, Suhas
Sartre, Gilles
Meyerholz, David K.
Théberge, Vanessa
Hallée, Stéphanie
Duperré, Anne-Marie
Del’Guidice, Thomas
Lepetit-Stoffaes, Jean-Pascal
Barbeau, Xavier
Guay, David
McCray, Paul B.
author_facet Krishnamurthy, Sateesh
Wohlford-Lenane, Christine
Kandimalla, Suhas
Sartre, Gilles
Meyerholz, David K.
Théberge, Vanessa
Hallée, Stéphanie
Duperré, Anne-Marie
Del’Guidice, Thomas
Lepetit-Stoffaes, Jean-Pascal
Barbeau, Xavier
Guay, David
McCray, Paul B.
author_sort Krishnamurthy, Sateesh
collection PubMed
description The delivery of biologic cargoes to airway epithelial cells is challenging due to the formidable barriers imposed by its specialized and differentiated cells. Among cargoes, recombinant proteins offer therapeutic promise but the lack of effective delivery methods limits their development. Here, we achieve protein and SpCas9 or AsCas12a ribonucleoprotein (RNP) delivery to cultured human well-differentiated airway epithelial cells and mouse lungs with engineered amphiphilic peptides. These shuttle peptides, non-covalently combined with GFP protein or CRISPR-associated nuclease (Cas) RNP, allow rapid entry into cultured human ciliated and non-ciliated epithelial cells and mouse airway epithelia. Instillation of shuttle peptides combined with SpCas9 or AsCas12a RNP achieves editing of loxP sites in airway epithelia of ROSA(mT/mG) mice. We observe no evidence of short-term toxicity with a widespread distribution restricted to the respiratory tract. This peptide-based technology advances potential therapeutic avenues for protein and Cas RNP delivery to refractory airway epithelial cells.
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spelling pubmed-68178252019-10-30 Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia Krishnamurthy, Sateesh Wohlford-Lenane, Christine Kandimalla, Suhas Sartre, Gilles Meyerholz, David K. Théberge, Vanessa Hallée, Stéphanie Duperré, Anne-Marie Del’Guidice, Thomas Lepetit-Stoffaes, Jean-Pascal Barbeau, Xavier Guay, David McCray, Paul B. Nat Commun Article The delivery of biologic cargoes to airway epithelial cells is challenging due to the formidable barriers imposed by its specialized and differentiated cells. Among cargoes, recombinant proteins offer therapeutic promise but the lack of effective delivery methods limits their development. Here, we achieve protein and SpCas9 or AsCas12a ribonucleoprotein (RNP) delivery to cultured human well-differentiated airway epithelial cells and mouse lungs with engineered amphiphilic peptides. These shuttle peptides, non-covalently combined with GFP protein or CRISPR-associated nuclease (Cas) RNP, allow rapid entry into cultured human ciliated and non-ciliated epithelial cells and mouse airway epithelia. Instillation of shuttle peptides combined with SpCas9 or AsCas12a RNP achieves editing of loxP sites in airway epithelia of ROSA(mT/mG) mice. We observe no evidence of short-term toxicity with a widespread distribution restricted to the respiratory tract. This peptide-based technology advances potential therapeutic avenues for protein and Cas RNP delivery to refractory airway epithelial cells. Nature Publishing Group UK 2019-10-28 /pmc/articles/PMC6817825/ /pubmed/31659165 http://dx.doi.org/10.1038/s41467-019-12922-y Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Krishnamurthy, Sateesh
Wohlford-Lenane, Christine
Kandimalla, Suhas
Sartre, Gilles
Meyerholz, David K.
Théberge, Vanessa
Hallée, Stéphanie
Duperré, Anne-Marie
Del’Guidice, Thomas
Lepetit-Stoffaes, Jean-Pascal
Barbeau, Xavier
Guay, David
McCray, Paul B.
Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia
title Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia
title_full Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia
title_fullStr Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia
title_full_unstemmed Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia
title_short Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia
title_sort engineered amphiphilic peptides enable delivery of proteins and crispr-associated nucleases to airway epithelia
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6817825/
https://www.ncbi.nlm.nih.gov/pubmed/31659165
http://dx.doi.org/10.1038/s41467-019-12922-y
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