Disease phenotype of a ferret CFTR-knockout model of cystic fibrosis
Xingshen Sun, Hongshu Sui, John T. Fisher, Ziying Yan, Xiaoming Liu, Hyung-Ju Cho, Nam Soo Joo, Yulong Zhang, Weihong Zhou, Yaling Yi, Joann M. Kinyon, Diana C. Lei-Butters, Michelle A. Griffin, Paul Naumann, Meihui Luo, Jill Ascher, Kai Wang, Timothy Frana, Jeffrey J. Wine, David K. Meyerholz, John F. Engelhardt
Xingshen Sun, Hongshu Sui, John T. Fisher, Ziying Yan, Xiaoming Liu, Hyung-Ju Cho, Nam Soo Joo, Yulong Zhang, Weihong Zhou, Yaling Yi, Joann M. Kinyon, Diana C. Lei-Butters, Michelle A. Griffin, Paul Naumann, Meihui Luo, Jill Ascher, Kai Wang, Timothy Frana, Jeffrey J. Wine, David K. Meyerholz, John F. Engelhardt
View: Text | PDF
Technical Advance

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

Abstract

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.

Authors

Xingshen Sun, Hongshu Sui, John T. Fisher, Ziying Yan, Xiaoming Liu, Hyung-Ju Cho, Nam Soo Joo, Yulong Zhang, Weihong Zhou, Yaling Yi, Joann M. Kinyon, Diana C. Lei-Butters, Michelle A. Griffin, Paul Naumann, Meihui Luo, Jill Ascher, Kai Wang, Timothy Frana, Jeffrey J. Wine, David K. Meyerholz, John F. Engelhardt

×

Figure 2

Primary organ pathologies observed in newborn CFTR–/– kits.

Options: View larger image (or click on image) Download as PowerPoint
Primary organ pathologies observed in newborn CFTR–/– kits. (A) H&am...
(A) H&E-stained sections of the pancreas. Eosinophilic zymogen material filled exocrine acini (asterisks) of the CFTR–/– pancreas. (B) H&E-stained sections of the liver and gall bladder (GB). Arrowheads mark the bile-filled canaliculi infrequently seen in all genotypes. (C) H&E-stained section of the lung demonstrating the type of lung lesions observed in CFTR–/– newborn kits. The CFTR–/– examples are from the same animal that passed stool in Figure 1C and died at 48 hours. Boxed regions are enlarged in the bottom row and demonstrate evidence of fibrin deposition, necrosis, bacteria, and/or inflammation. (D) Blood chemistries for ALT, total bilirubin (Tbil), and cholesterol (CHOL) in animals of the indicated genotypes (WT, CFTR+/+ and CFTR+/–; CF, CFTR–/–). CF kits were divided into 2 groups with and without MI. BL, below limits of detection. Blood was drawn at the time of euthanasia. Values depict the mean ± SEM (n = 5–9 animals in each group) (see Supplemental Figure 3 for additional blood chemistry data). Scale bar: 200 μm (A, top row, B, top row, and C, bottom left and right panels); 25 μm (A, bottom row); 50 μm (B, bottom row); 500 μm (C, top row); 100 μm (C, bottom center panel).