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

Sateesh Krishnamurthy; Christine Wohlford-Lenane; Suhas Kandimalla; Gilles Sartre; David K Meyerholz; Vanessa Théberge; Stéphanie Hallée; Anne-Marie Duperré; Thomas Del'Guidice; Jean-Pascal Lepetit-Stoffaes; Xavier Barbeau; David Guay; Paul B McCray Jr

doi:10.1038/s41467-019-12922-y

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

Journal article

Open access

Peer reviewed

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

Sateesh Krishnamurthy, Christine Wohlford-Lenane, Suhas Kandimalla, Gilles Sartre, David K Meyerholz, Vanessa Théberge, Stéphanie Hallée, Anne-Marie Duperré, Thomas Del'Guidice, Jean-Pascal Lepetit-Stoffaes, …

Nature communications, Vol.10(1), 4906

10/28/2019

DOI: 10.1038/s41467-019-12922-y

PMCID: PMC6817825

PMID: 31659165

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https://doi.org/10.1038/s41467-019-12922-yView

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Abstract

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 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.

Animals

Bacterial Proteins - genetics

Bacterial Proteins - metabolism

Bronchi - cytology

Bronchi - metabolism

Drug Delivery Systems - methods

Endonucleases - genetics

Endonucleases - metabolism

Epithelial Cells - metabolism

Genetic Therapy

Humans

Lung - metabolism

Lung Diseases - genetics

Lung Diseases - metabolism

Lung Diseases - therapy

Mice

Peptides - administration & dosage

Peptides - genetics

Peptides - metabolism

Ribonucleoproteins - genetics

Ribonucleoproteins - metabolism

Swine

Details

Title: Subtitle: Engineered amphiphilic peptides enable delivery of proteins and CRISPR-associated nucleases to airway epithelia
Creators: Sateesh Krishnamurthy - University of Iowa
Christine Wohlford-Lenane - University of Iowa
Suhas Kandimalla - University of Iowa
Gilles Sartre - University of Iowa
David K Meyerholz - University of Iowa
Vanessa Théberge - Feldan Therapeutics, Québec, QC, Canada.
Stéphanie Hallée - Feldan Therapeutics, Québec, QC, Canada.
Anne-Marie Duperré - Feldan Therapeutics, Québec, QC, Canada
Thomas Del'Guidice - Feldan Therapeutics, Québec, QC, Canada
Jean-Pascal Lepetit-Stoffaes - Feldan Therapeutics, Québec, QC, Canada
Xavier Barbeau - Feldan Therapeutics, Québec, QC, Canada.
David Guay - Feldan Therapeutics, Québec, QC, Canada.
Paul B McCray Jr - University of Iowa
Resource Type: Journal article
Publication Details: Nature communications, Vol.10(1), 4906
DOI: 10.1038/s41467-019-12922-y
PMID: 31659165
PMCID: PMC6817825
NLM abbreviation: Nat Commun
ISSN: 2041-1723
eISSN: 2041-1723
Grant note: P30 ES005605 / NIEHS NIH HHS P01 HL091842 / NHLBI NIH HHS P01 HL051670 / NHLBI NIH HHS UG3 HL147366 / NHLBI NIH HHS P30 DK054759 / NIDDK NIH HHS
Language: English
Date published: 10/28/2019
Academic Unit: Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Pathology; Internal Medicine
Record Identifier: 9984186775002771

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