Cystic fibrosis transmembrane conductance regulator with a shortened R domain rescues the intestinal phenotype of CFTR-/- mice

Lynda S Ostedgaard; David K Meyerholz; Daniel W Vermeer; Philip H Karp; Lindsey Schneider; Curt D Sigmund; Michael J Welsh

doi:10.1073/pnas.1019752108

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Cystic fibrosis transmembrane conductance regulator with a shortened R domain rescues the intestinal phenotype of CFTR-/- mice

Journal article

Open access

Peer reviewed

Cystic fibrosis transmembrane conductance regulator with a shortened R domain rescues the intestinal phenotype of CFTR-/- mice

Lynda S Ostedgaard, David K Meyerholz, Daniel W Vermeer, Philip H Karp, Lindsey Schneider, Curt D Sigmund and Michael J Welsh

Proceedings of the National Academy of Sciences - PNAS, Vol.108(7), pp.2921-2926

02/15/2011

DOI: 10.1073/pnas.1019752108

PMCID: PMC3041073

PMID: 21285372

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Abstract

Gene transfer could provide a novel therapeutic approach for cystic fibrosis (CF), and adeno-associated virus (AAV) is a promising vector. However, the packaging capacity of AAV limits inclusion of the full-length cystic fibrosis transmembrane conductance regulator (CFTR) cDNA together with other regulatory and structural elements. To overcome AAV size constraints, we recently developed a shortened CFTR missing the N-terminal portion of the R domain (residues 708-759, CFTRΔR) and found that it retained regulated anion channel activity in vitro. To test the hypothesis that CFTRΔR could correct in vivo defects, we generated CFTR(-/-) mice bearing a transgene with a fatty acid binding protein promoter driving expression of human CFTRΔR in the intestine (CFTR(-/-);TgΔR). We found that intestinal crypts of CFTR(-/-);TgΔR mice expressed CFTRΔR and the intestine appeared histologically similar to that of WT mice. Moreover, like full-length CFTR transgene, the CFTRΔR transgene produced CFTR Cl(-) currents and rescued the CFTR(-/-) intestinal phenotype. These results indicate that the N-terminal part of the CFTR R domain is dispensable for in vivo intestinal physiology. Thus, CFTRΔR may have utility for AAV-mediated gene transfer in CF.

Cystic Fibrosis - therapy

Immunohistochemistry

Electrophoresis, Polyacrylamide Gel

Humans

Electrophysiology

Intestines - metabolism

Protein Structure, Tertiary - genetics

Dependovirus

Genetic Vectors - genetics

Mice, Knockout

Intestines - anatomy & histology

Animals

Cystic Fibrosis - genetics

Cystic Fibrosis Transmembrane Conductance Regulator - genetics

Mice

Genetic Therapy - methods

Details

Title: Subtitle: Cystic fibrosis transmembrane conductance regulator with a shortened R domain rescues the intestinal phenotype of CFTR-/- mice
Creators: Lynda S Ostedgaard - Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
David K Meyerholz
Daniel W Vermeer
Philip H Karp
Lindsey Schneider
Curt D Sigmund
Michael J Welsh
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.108(7), pp.2921-2926
DOI: 10.1073/pnas.1019752108
PMID: 21285372
PMCID: PMC3041073
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences; United States
Grant note: HL091842 / NHLBI NIH HHS P01 HL091842 / NHLBI NIH HHS P01 HL051670 / NHLBI NIH HHS HL51670 / NHLBI NIH HHS Howard Hughes Medical Institute
Language: English
Date published: 02/15/2011
Academic Unit: Neurology; Molecular Physiology and Biophysics; Pulmonary, Critical Care, and Occupational Medicine; Pathology; Neuroscience and Pharmacology; Neurosurgery; Internal Medicine
Record Identifier: 9984020619202771

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