Journal article
Lentiviral vectors transduce lung stem cells without disrupting plasticity
Molecular therapy. Nucleic acids, Vol.25, pp.293-301
09/03/2021
DOI: 10.1016/j.omtn.2021.06.010
PMCID: PMC8379527
PMID: 34458011
Abstract
Life-long expression of a gene therapy agent likely requires targeting stem cells. Here we ask the question: does viral vector transduction or ectopic expression of a therapeutic transgene preclude airway stem cell function? We used a lentiviral vector containing a GFP or cystic fibrosis transmembrane conductance regulator (CFTR) transgene to transduce primary airway basal cells from human cystic fibrosis (CF) or non-CF lung donors and monitored expression and function after differentiation. Ussing chamber measurements confirmed CFTR-dependent chloride channel activity in CF donor cells. Immunostaining, quantitative real-time PCR, and single-cell sequencing analysis of cell-type markers indicated that vector transduction or CFTR expression does not alter the formation of pseudostratified, fully differentiated epithelial cell cultures or cell type distribution. These results have important implications for use of gene addition or gene editing strategies as life-long curative approaches for lung genetic diseases.
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Does lentivirus-mediated transgene expression alter airway stem cell function? Based on morphology, physiology, and single-cell RNA sequencing, the authors conclude that lentivirus-mediated phenotypic correction has remarkably little impact on global gene expression or stem cell plasticity. These results have critical implications for lentivirus as a life-long curative strategy.
Details
- Title: Subtitle
- Lentiviral vectors transduce lung stem cells without disrupting plasticity
- Creators
- Ashley L. Cooney - University of IowaAndrew L. Thurman - University of IowaPaul B. McCray - University of IowaAlejandro A. Pezzulo - University of IowaPatrick L. Sinn - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Molecular therapy. Nucleic acids, Vol.25, pp.293-301
- DOI
- 10.1016/j.omtn.2021.06.010
- PMID
- 34458011
- PMCID
- PMC8379527
- NLM abbreviation
- Mol Ther Nucleic Acids
- ISSN
- 2162-2531
- eISSN
- 2162-2531
- Publisher
- Elsevier Inc
- Grant note
- DOI: 10.13039/100000002, name: National Institutes of Health, award: P01 HL091842, P01 HL152960, P01 HL51670, R01 HL133089; DOI: 10.13039/100000897, name: Cystic Fibrosis Foundation, award: COONEY18F0, SINN19XX0; DOI: 10.13039/100011352, name: Center for Gene Therapy, University of Iowa, award: DK54759
- Language
- English
- Date published
- 09/03/2021
- Academic Unit
- Pulmonary, Critical Care, and Occupational Medicine; Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Internal Medicine
- Record Identifier
- 9984297425602771
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