Nature Biotechnology
20, 47 - 52 (2002)
doi:10.1038/nbt0102-47
Partial correction of endogenous  F508 CFTR in human cystic fibrosis airway epithelia by spliceosome-mediated RNA trans-splicingXiaoming Liu1, 6, Qinshi Jiang1, 6, S. Gary Mansfield4, M. Puttaraju4, Yulong Zhang1, 3, Weihong Zhou1, Jonathan A. Cohn5, Mariano A. Garcia-Blanco4, Lloyd G. Mitchell4
& John F. Engelhardt1, 2, 31
Department of Anatomy and Cell Biology, College of Medicine, The University of Iowa, Iowa City, IA 52242. 2
Department of Internal Medicine, College of Medicine, The University of Iowa, Iowa City, IA 52242. 3
The Center for Gene Therapy of Cystic Fibrosis and Other Genetic Diseases, College of Medicine, The University of Iowa, Iowa City, IA 52242. 4
Intronn Inc, Raleigh, NC 27606. 5
Department of Medicine, Duke University Medical Center, Durham, NC 27710. 6
These authors contributed equally to this work.
Correspondence should be addressed to John F. Engelhardt john-engelhardt@uiowa.eduSpliceosome-mediated RNA trans-splicing (SMaRT) was investigated as a means for functionally correcting endogenous  F508 cystic fibrosis transmembrane conductance regulator (CFTR) transcripts using in vitro human cystic fibrosis (CF) polarized airway epithelia and in vivo human CF bronchial xenografts. Recombinant adenovirus (Ad.CFTR-PTM) encoding a pre-therapeutic molecule (PTM) targeted to CFTR intron 9 corrected transepithelial cyclic AMP (cAMP)-sensitive short-circuit current (Isc) in F508 homozygous epithelia to a level 16% of that observed in normal human bronchial epithelia. Molecular analyses using RT-PCR and western blotting confirmed SMaRT-mediated partial correction of endogenous F508 messenger RNA (mRNA) transcripts and protein. In an in vivo model of F508 CF airway epithelia, human CF bronchial xenografts infected with Ad.CFTR-PTM also demonstrated partial correction of CFTR-mediated Cl- permeability at a level 22% of that seen in non-CF xenografts. These results provide functional evidence for SMaRT-mediated repair of mutant endogenous CFTR mRNA in intact polarized CF airway epithelial models.
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