i***R
发帖数: 663 | 1 这不知道得多大的场地做实验。
Sci Transl Med 16 March 2011:
Vol. 3, Issue 74, p. 74ra24
DOI: 10.1126/scitranslmed.3001868
RESEARCH ARTICLE
CYSTIC FIBROSIS
The ΔF508 Mutation Causes CFTR Misprocessing and Cystic Fibrosis–Like
Disease in Pigs
Lynda S. Ostedgaard1,*, David K. Meyerholz2,*, Jeng-Haur Chen1,3,*,
Alejandro A. Pezzulo1, Philip H. Karp1,3, Tatiana Rokhlina1, Sarah E.
Ernst1, Robert A. Hanfland4, Leah R. Reznikov1, Paula S. Ludwig1, Mark
P. Rogan1, Greg J. Davis1, Cassie L. Dohrn2, Christine Wohlford-Lenane5,
Peter J. Taft1, Michael V. Rector1, Emma Hornick1, Boulos S. Nassar1,
Melissa Samuel6, Yuping Zhang1, Sandra S. Richter2, Aliye Uc5, Joel
Shilyansky4, Randall S. Prather6, Paul B. McCray Jr.5, Joseph Zabner1,
Michael J. Welsh1,3,† and David A. Stoltz1,†
+ Author Affiliations
1Department of Internal Medicine, Roy J. and Lucille A. Carver College
of Medicine, University of Iowa, Iowa City, IA 52242, USA.
2Department of Pathology, Roy J. and Lucille A. Carver College of
Medicine, University of Iowa, Iowa City, IA 52242, USA.
3Howard Hughes Medical Institute, Roy J. and Lucille A. Carver College
of Medicine, University of Iowa, Iowa City, IA 52242, USA.
4Department of Surgery, Roy J. and Lucille A. Carver College of
Medicine, University of Iowa, Iowa City, IA 52242, USA.
5Department of Pediatrics, Roy J. and Lucille A. Carver College of
Medicine, University of Iowa, Iowa City, IA 52242, USA.
6Division of Animal Sciences, University of Missouri, Columbia, MO
65211, USA.
†To whom correspondence should be addressed. E-mail: michael-
w***[email protected] (M.J.W.); d**********[email protected] (D.A.S.)
ABSTRACT
Cystic fibrosis (CF) is an autosomal recessive disease caused by
mutations in the gene encoding the cystic fibrosis transmembrane
conductance regulator (CFTR) anion channel. The most common CF-
associated mutation is ΔF508, which deletes a phenylalanine in position
508. In vitro studies indicate that the resultant protein, CFTR-ΔF508,
is misprocessed, although the in vivo consequences of this mutation
remain uncertain. To better understand the effects of the ΔF508 mutation
in vivo, we produced CFTRΔF508/ΔF508 pigs. Our biochemical,
immunocytochemical, and electrophysiological data on CFTR-ΔF508 in
newborn pigs paralleled in vitro predictions. They also indicated that
CFTRΔF508/ΔF508 airway epithelia retain a small residual CFTR
conductance, with maximal stimulation producing ~6% of wild-type
function. Cyclic adenosine 3′,5′-monophosphate (cAMP) agonists were less
potent at stimulating current in CFTRΔF508/ΔF508 epithelia, suggesting
that quantitative tests of maximal anion current may overestimate
transport under physiological conditions. Despite residual CFTR
function, four older CFTRΔF508/ΔF508 pigs developed lung disease similar
to human CF. These results suggest that this limited CFTR activity is
insufficient to prevent lung or gastrointestinal disease in CF pigs.
These data also suggest that studies of recombinant CFTR-ΔF508
misprocessing predict in vivo behavior, which validates its use in
biochemical and drug discovery experiments. These findings help
elucidate the molecular pathogenesis of the common CF mutation and will
guide strategies for developing new therapeutics.
↵* These authors contributed equally to this work. |
|
