Journal article
Increased CFTR expression and function from an optimized lentiviral vector for cystic fibrosis gene therapy
Molecular therapy. Methods & clinical development, Vol.21, pp.94-106
06/11/2021
DOI: 10.1016/j.omtm.2021.02.020
PMCID: PMC7973238
PMID: 33768133
Abstract
Despite significant advances in cystic fibrosis (CF) treatments, a one-time treatment for this life-shortening disease remains elusive. Stable complementation of the disease-causing mutation with a normal copy of the CF transmembrane conductance regulator (CFTR) gene fulfills that goal. Integrating lentiviral vectors are well suited for this purpose, but widespread airway transduction in humans is limited by achievable titers and delivery barriers. Since airway epithelial cells are interconnected through gap junctions, small numbers of cells expressing supraphysiologic levels of CFTR could support sufficient channel function to rescue CF phenotypes. Here, we investigated promoter choice and CFTR codon optimization (coCFTR) as strategies to regulate CFTR expression. We evaluated two promoters-phosphoglycerate kinase (PGK) and elongation factor 1-alpha (EF1 alpha)-that have been safely used in clinical trials. We also compared the wild-type human CFTR sequence to three alternative coCFTR sequences generated by different algorithms. With the use of the CFTR-mediated anion current in primary human CF airway epithelia to quantify channel expression and function, we determined that EF1 alpha produced greater currents than PGK and identified a coCFTR sequence that conferred significantly increased functional CFTR expression. Optimized promoter and CFTR sequences advance lentiviral vectors toward CF gene therapy clinical trials.
Details
- Title: Subtitle
- Increased CFTR expression and function from an optimized lentiviral vector for cystic fibrosis gene therapy
- Creators
- Laura I Marquez Loza - University of IowaAshley L. Cooney - University of IowaQian Dong - University of IowaChristoph O. Randak - University of IowaStefano Rivella - Children's Hospital of PhiladelphiaPatrick L. Sinn - University of IowaPaul B. McCray - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Molecular therapy. Methods & clinical development, Vol.21, pp.94-106
- Publisher
- Elsevier
- DOI
- 10.1016/j.omtm.2021.02.020
- PMID
- 33768133
- PMCID
- PMC7973238
- ISSN
- 2329-0501
- eISSN
- 2329-0501
- Number of pages
- 13
- Grant note
- Cystic Fibrosis Foundation; Italian Cystic Fibrosis Research Foundation F31 HL152500; UG3 HL147366; P01 HL51670; P01 HL091842; P01 HL152960; R01DK107489; T32 GM007337 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA DK54759 / University of Iowa Center for Gene Therapy Roy J. Carver Chair in Pulmonary Research
- Language
- English
- Date published
- 06/11/2021
- Academic Unit
- Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Internal Medicine
- Record Identifier
- 9984297433102771
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