Hepatobiliary disease causes significant morbidity in people with cystic fibrosis (CF), yet this problem remains understudied. We previously found that newborn CF pigs have microgallbladders with significant luminal obstruction in the absence of infection and consistent inflammation. In this study, we sought to better understand the early pathogenesis of CF pig gallbladder disease. We hypothesized that loss of CFTR would impair gallbladder epithelium anion/liquid secretion and increase mucin production. CFTR was expressed apically in non-CF pig gallbladder epithelium but was absent in CF. CF pig gallbladders lacked cAMP-stimulated anion transport. Using a novel gallbladder epithelial organoid model, we found that Cl− or HCO3− was sufficient for non-CF organoid swelling. This response was absent for non-CF organoids in Cl−/HCO3−-free conditions and in CF. Single-cell RNA-sequencing revealed a single epithelial cell type in non-CF gallbladders that coexpressed CFTR, MUC5AC, and MUC5B. Despite CF gallbladders having increased luminal MUC5AC and MUC5B accumulation, there was no significant difference in the epithelial expression of gel-forming mucins between non-CF and CF pig gallbladders. In conclusion, these data suggest that loss of CFTR-mediated anion transport and fluid secretion contribute to microgallbladder development and luminal mucus accumulation in CF.
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We thank M. Abou Alaiwa, A. Comellas, H. de Jonge, M. Duffey, P. McCray, T. Moninger, F. Sampaziotis, M. Welsh, J. Zabner, the University of Iowa Office of Animal Resources, the University of Iowa Genomics Division, and the University of Iowa Comparative Pathology Lab for excellent assistance, advice, and discussions. This work was supported, in part, by NIH (HL091842, GM007337, and HL007638) and the Cystic Fibrosis Foundation (CFF Iowa Research Development Program).
Conflict of interest
The University of Iowa Research Foundation has licensed materials and technologies related to CF pigs to Exemplar Genetics. DAS is a coinventor of CF pigs.
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Zarei, K., Stroik, M.R., Gansemer, N.D. et al. Early pathogenesis of cystic fibrosis gallbladder disease in a porcine model. Lab Invest (2020). https://doi.org/10.1038/s41374-020-0474-8