Journal article
Detargeting Lentiviral-Mediated CFTR Expression in Airway Basal Cells Using miR-106b
Genes, Vol.11(10), pp.1-20
10/06/2020
DOI: 10.3390/genes11101169
PMCID: PMC7601932
PMID: 33036232
Abstract
Lentiviral-mediated integration of a
transgene cassette into airway basal cells is a strategy being considered for cystic fibrosis (CF) cell-based therapies. However,
expression is highly regulated in differentiated airway cell types and a subset of intermediate basal cells destined to differentiate. Since basal stem cells typically do not express CFTR, suppressing the
expression from the lentiviral vector in airway basal cells may be beneficial for maintaining their proliferative capacity and multipotency. We identified miR-106b as highly expressed in proliferating airway basal cells and extinguished in differentiated columnar cells. Herein, we developed lentiviral vectors with the miR-106b-target sequence (miRT) to both study miR-106b regulation during basal cell differentiation and detarget CFTR expression in basal cells. Given that miR-106b is expressed in the 293T cells used for viral production, obstacles of viral genome integrity and titers were overcome by creating a 293T-B2 cell line that inducibly expresses the RNAi suppressor B2 protein from flock house virus. While miR-106b vectors effectively detargeted reporter gene expression in proliferating basal cells and following differentiation in the air-liquid interface and organoid cultures, the CFTR-miRT vector produced significantly less CFTR-mediated current than the non-miR-targeted CFTR vector following transduction and differentiation of CF basal cells. These findings suggest that miR-106b is expressed in certain airway cell types that contribute to the majority of CFTR anion transport in airway epithelium.
Details
- Title: Subtitle
- Detargeting Lentiviral-Mediated CFTR Expression in Airway Basal Cells Using miR-106b
- Creators
- Soon H Choi - Roy J. and Lucille A. Carver College of MedicineRosie E Reeves - University of IowaGuillermo S Romano Ibarra - University of IowaThomas J Lynch - University of IowaWeam S Shahin - University of IowaZehua Feng - University of IowaGrace N Gasser - Roy J. and Lucille A. Carver College of MedicineMichael C Winter - University of IowaT Idil Apak Evans - University of IowaXiaoming Liu - University of IowaMeihui Luo - University of IowaYulong Zhang - University of IowaDavid A Stoltz - University of IowaEric J Devor - University of IowaZiying Yan - Roy J. and Lucille A. Carver College of MedicineJohn F Engelhardt - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Genes, Vol.11(10), pp.1-20
- DOI
- 10.3390/genes11101169
- PMID
- 33036232
- PMCID
- PMC7601932
- NLM abbreviation
- Genes (Basel)
- ISSN
- 2073-4425
- eISSN
- 2073-4425
- Grant note
- P30 ES005605 / NIEHS NIH HHS P30 DK054759 / NIH HHS T32 GM007337 / NIH HHS R01 DK047967 / NIH HHS T32 GM007337 / NIGMS NIH HHS T32 HL007638 / NIH HHS P01 HL152960 / NIH HHS P30 DK054759 / NIDDK NIH HHS
- Language
- English
- Date published
- 10/06/2020
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Molecular Physiology and Biophysics; Pulmonary, Critical Care, and Occupational Medicine; Anatomy and Cell Biology; Radiation Oncology; Obstetrics and Gynecology; Cardiothoracic Surgery; Internal Medicine
- Record Identifier
- 9984284338202771
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