Disease phenotype of a ferret CFTR-knockout model of cystic fibrosis.

Cystic fibrosis (CF) is a recessive disease that affects multiple organs. It is caused by mutations in CFTR. Animal modeling of this disease has been challenging, with species- and strain-specific differences in organ biology and CFTR function influencing the emergence of disease pathology. Here, we report the phenotype of a CFTR-knockout ferret model of CF. Neonatal CFTR-knockout ferrets demonstrated many of the characteristics of human CF disease, including defective airway chloride transport and submucosal gland fluid secretion; variably penetrant meconium ileus (MI); pancreatic, liver, and vas deferens disease; and a predisposition to lung infection in the early postnatal period. Severe malabsorption by the gastrointestinal (GI) tract was the primary cause of death in CFTR-knockout kits that escaped MI. Elevated liver function tests in CFTR-knockout kits were corrected by oral administration of ursodeoxycholic acid, and the addition of an oral proton-pump inhibitor improved weight gain and survival. To overcome the limitations imposed by the severe intestinal phenotype, we cloned 4 gut-corrected transgenic CFTR-knockout kits that expressed ferret CFTR specifically in the intestine. One clone passed feces normally and demonstrated no detectable ferret CFTR expression in the lung or liver. The animals described in this study are likely to be useful tools for dissecting CF disease pathogenesis and developing treatments.

[1]  J. Gustafson,et al.  Cystic Fibrosis , 2009, Journal of the Iowa Medical Society.

[2]  M. Egan How useful are cystic fibrosis mouse models , 2009 .

[3]  A. Hofmann Bile acids: Trying to understand their chemistry and biology with the hope of helping patients , 2009, Hepatology.

[4]  G. Lukács,et al.  N-glycans are direct determinants of CFTR folding and stability in secretory and endocytic membrane traffic , 2009, The Journal of cell biology.

[5]  D. Meyerholz,et al.  Disruption of the CFTR Gene Produces a Model of Cystic Fibrosis in Newborn Pigs , 2008, Science.

[6]  Xingshen Sun,et al.  Adeno-associated virus-targeted disruption of the CFTR gene in cloned ferrets. , 2008, The Journal of clinical investigation.

[7]  Jae Young Choi,et al.  Synergistic airway gland mucus secretion in response to vasoactive intestinal peptide and carbachol is lost in cystic fibrosis. , 2007, The Journal of clinical investigation.

[8]  M. Welsh,et al.  Processing and function of CFTR-ΔF508 are species-dependent , 2007, Proceedings of the National Academy of Sciences.

[9]  A. Neucker,et al.  Effects of a proton-pump inhibitor in cystic fibrosis , 2007 .

[10]  A. Chakravarti,et al.  Relative contribution of genetic and nongenetic modifiers to intestinal obstruction in cystic fibrosis. , 2006, Gastroenterology.

[11]  Qi Zhou,et al.  Cloned ferrets produced by somatic cell nuclear transfer. , 2006, Developmental biology.

[12]  Michael R Knowles,et al.  Genetic modifiers of lung disease in cystic fibrosis. , 2005, The New England journal of medicine.

[13]  H. Verkade,et al.  Fat absorption in cystic fibrosis mice is impeded by defective lipolysis and post-lipolytic events. , 2005, American journal of physiology. Gastrointestinal and liver physiology.

[14]  M. Welsh,et al.  Lysozyme secretion by submucosal glands protects the airway from bacterial infection. , 2005, American journal of respiratory cell and molecular biology.

[15]  A. Verkman,et al.  Submucosal gland dysfunction as a primary defect in cystic fibrosis , 2005, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[16]  J. Szamatowicz,et al.  Cystic fibrosis as a cause of infertility. , 2004, Reproductive biology.

[17]  M. Proesmans,et al.  Omeprazole, a proton pump inhibitor, improves residual steatorrhoea in cystic fibrosis patients treated with high dose pancreatic enzymes , 2003, European Journal of Pediatrics.

[18]  A. Manning,et al.  The normal upper gastrointestinal examination in the ferret. , 2003, Veterinary radiology & ultrasound : the official journal of the American College of Veterinary Radiology and the International Veterinary Radiology Association.

[19]  N. Joo,et al.  Optical method for quantifying rates of mucus secretion from single submucosal glands. , 2001, American journal of physiology. Lung cellular and molecular physiology.

[20]  J. Widdicombe,et al.  A comparative study of mammalian tracheal mucous glands , 2000, Journal of anatomy.

[21]  J. Cassiman,et al.  Morphological changes in the vas deferens and expression of the cystic fibrosis transmembrane conductance regulator (CFTR) in control, ΔF508 and knock‐out CFTR mice during postnatal life , 2000, Molecular reproduction and development.

[22]  B. Strandvik,et al.  Natural history of liver disease in cystic fibrosis , 1999, Hepatology.

[23]  D. Gaillard,et al.  Normal vas deferens in fetuses with cystic fibrosis. , 1997, The Journal of urology.

[24]  P. McCray,et al.  Ursodeoxycholic Acid Improves Cholestasis in Infants with Cystic Fibrosis , 1997, The Annals of pharmacotherapy.

[25]  J. Yankaskas,et al.  In vivo analysis of fluid transport in cystic fibrosis airway epithelia of bronchial xenografts. , 1996, The American journal of physiology.

[26]  J. Whitsett,et al.  Correction of lethal intestinal defect in a mouse model of cystic fibrosis by human CFTR. , 1994, Science.

[27]  K. J. Mysels,et al.  Bile acid solubility and precipitation in vitro and in vivo: the role of conjugation, pH, and Ca2+ ions. , 1992, Journal of lipid research.

[28]  M. Tanner,et al.  Prevalence of liver disease in cystic fibrosis. , 1991, Archives of disease in childhood.

[29]  M. Lentze,et al.  Effects of ursodeoxycholic acid treatment on nutrition and liver function in patients with cystic fibrosis and longstanding cholestasis. , 1990, Gut.

[30]  L. Tsui,et al.  Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. , 1989, Science.

[31]  J. Widdicombe,et al.  Quantitation of the secretory cells of the ferret tracheobronchial tree. , 1986, Journal of anatomy.

[32]  C. Roy,et al.  Bile Acid Metabolism in Children with Cystic Fibrosis , 1985, Acta paediatrica Scandinavica. Supplement.

[33]  C. Plopper,et al.  Ultrastructure of the nonciliated bronchiolar epithelial (Clara) cell of mammalian lung. III. A study of man with comparison of 15 mammalian species. , 1980, Experimental lung research.

[34]  E. Roma,et al.  Intestinal bile salts in cystic fibrosis: studies in the patient and experimental animal. , 1979, Archives of disease in childhood.

[35]  C. Roy,et al.  Malabsorption of bile acids in children with cystic fibrosis. , 1973, The New England journal of medicine.

[36]  J. Esterly,et al.  Cystic fibrosis of the pancreas. Morphologic findings in infants with and without diagnostic pancreatic lesions. , 1973, Archives of pathology.

[37]  J. A. Fraser Roberts,et al.  Fibrocystic disease of the pancreas. A congenital disorder of mucus production—mucosis , 1953 .

[38]  G. Downey,et al.  Cystic fibrosis mouse models. , 2007, American journal of respiratory cell and molecular biology.

[39]  N. Joo,et al.  Submucosal glands and airway defense. , 2004, Proceedings of the American Thoracic Society.

[40]  J. Widdicombe,et al.  The distribution and structure of cells in the tracheal epithelium of the mouse , 2004, Cell and Tissue Research.

[41]  D. Porteous,et al.  CFTR and calcium-activated chloride currents in pancreatic duct cells of a transgenic CF mouse. , 1994, The American journal of physiology.

[42]  P. Quinn Clinical veterinary microbiology , 1994 .

[43]  James M. Wilson,et al.  Submucosal glands are the predominant site of CFTR expression in the human bronchus , 1992, Nature Genetics.

[44]  Esterly,et al.  Pathology of cystic fibrosis review of the literature and comparison with 146 autopsied cases. , 1975, Perspectives in pediatric pathology.

[45]  D. Andersen Cystic fibrosis of the pancreas. , 1958, Journal of chronic diseases.

[46]  Ollett Ws A method for staining both gram-positive and gram-negative bacteria in sections. , 1947 .