Optimization of hCFTR lung expression in mice using DNA nanoparticles. Mol Ther 2012 Jan;20(1):63-72
Date
09/29/2011Pubmed ID
21952168Pubmed Central ID
PMC3255587DOI
10.1038/mt.2011.196Scopus ID
2-s2.0-84856961942 (requires institutional sign-in at Scopus site) 31 CitationsAbstract
Efficient and prolonged human cystic fibrosis transmembrane conductance regulator (hCFTR) expression is a major goal for cystic fibrosis (CF) lung therapy. A hCFTR expression plasmid was optimized as a payload for compacted DNA nanoparticles formulated with polyethylene glycol (PEG)-substituted 30-mer lysine peptides. A codon-optimized and CpG-reduced hCFTR synthetic gene (CO-CFTR) was placed in a polyubiquitin C expression plasmid. Compared to hCFTR complementary DNA (cDNA), CO-CFTR produced a ninefold increased level of hCFTR protein in transfected HEK293 cells and, when compacted as DNA nanoparticles, produced a similar improvement in lung mRNA expression in Balb/c and fatty acid binding protein promoter (FABP) CF mice, although expression duration was transient. Various vector modifications were tested to extend duration of CO-CFTR expression. A novel prolonged expression (PE) element derived from the bovine growth hormone (BGH) gene 3' flanking sequence produced prolonged expression of CO-CFTR mRNA at biologically relevant levels. A time course study in the mouse lung revealed that CO-CFTR mRNA did not change significantly, with CO-CFTR/mCFTR geometric mean ratios of 94% on day 2, 71% on day 14, 53% on day 30, and 14% on day 59. Prolonged CO-CFTR expression is dependent on the orientation of the PE element and its transcription, is not specific to the UbC promoter, and is less dependent on other vector backbone elements.
Author List
Padegimas L, Kowalczyk TH, Adams S, Gedeon CR, Oette SM, Dines K, Hyatt SL, Sesenoglu-Laird O, Tyr O, Moen RC, Cooper MJAuthor
Samuel J. Adams MD Assistant Professor in the Neurology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
3' Flanking RegionAdministration, Intranasal
Animals
Cystic Fibrosis
Cystic Fibrosis Transmembrane Conductance Regulator
DNA
Disease Models, Animal
Fatty Acid-Binding Proteins
Gene Expression
Gene Expression Regulation
Gene Order
Genetic Therapy
Genetic Vectors
HEK293 Cells
Humans
Lung
Mice
Mice, Inbred BALB C
Mice, Transgenic
Nanoconjugates
Promoter Regions, Genetic
RNA, Messenger
Transcription, Genetic