Ferret models of alpha-1 antitrypsin deficiency develop lung and liver disease
Nan He, Xiaoming Liu, Amber R. Vegter, T. Idil A. Evans, Jaimie S. Gray, Junfeng Guo, Shashanna R. Moll, Lydia J. Guo, Meihui Luo, Ningxia Ma, Xingshen Sun, Bo Liang, Ziying Yan, Zehua Feng, Lisi Qi, Arnav S. Joshi, Weam Shahin, Yaling Yi, Katherine N. Gibson-Corley, Eric A. Hoffman, Kai Wang, Christian Mueller, John F. Engelhardt, Bradley H. Rosen
Nan He, Xiaoming Liu, Amber R. Vegter, T. Idil A. Evans, Jaimie S. Gray, Junfeng Guo, Shashanna R. Moll, Lydia J. Guo, Meihui Luo, Ningxia Ma, Xingshen Sun, Bo Liang, Ziying Yan, Zehua Feng, Lisi Qi, Arnav S. Joshi, Weam Shahin, Yaling Yi, Katherine N. Gibson-Corley, Eric A. Hoffman, Kai Wang, Christian Mueller, John F. Engelhardt, Bradley H. Rosen
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Research Article Pulmonology

Ferret models of alpha-1 antitrypsin deficiency develop lung and liver disease

Abstract

Alpha-1 antitrypsin deficiency (AATD) is the most common genetic cause and risk factor for chronic obstructive pulmonary disease, but the field lacks a large-animal model that allows for longitudinal assessment of pulmonary function. We hypothesized that ferrets would model human AATD-related lung and hepatic disease. AAT-knockout (AAT-KO) and PiZZ (E342K, the most common mutation in humans) ferrets were generated and compared with matched controls using custom-designed flexiVent modules to perform pulmonary function tests, quantitative computed tomography (QCT), bronchoalveolar lavage (BAL) proteomics, and alveolar morphometry. Complete loss of AAT (AAT-KO) led to increased pulmonary compliance and expiratory airflow limitation, consistent with obstructive lung disease. QCT and morphometry confirmed emphysema and airspace enlargement, respectively. Pathway analysis of BAL proteomics data revealed inflammatory lung disease and impaired cellular migration. The PiZ mutation resulted in altered AAT protein folding in the liver, hepatic injury, and reduced plasma concentrations of AAT, and PiZZ ferrets developed obstructive lung disease. In summary, AAT-KO and PiZZ ferrets model the progressive obstructive pulmonary disease seen in AAT-deficient patients and may serve as a platform for preclinical testing of therapeutics including gene therapy.

Authors

Nan He, Xiaoming Liu, Amber R. Vegter, T. Idil A. Evans, Jaimie S. Gray, Junfeng Guo, Shashanna R. Moll, Lydia J. Guo, Meihui Luo, Ningxia Ma, Xingshen Sun, Bo Liang, Ziying Yan, Zehua Feng, Lisi Qi, Arnav S. Joshi, Weam Shahin, Yaling Yi, Katherine N. Gibson-Corley, Eric A. Hoffman, Kai Wang, Christian Mueller, John F. Engelhardt, Bradley H. Rosen

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Figure 5

BAL from AAT-KO ferrets has reduced NE inhibitory capacity, and secreted proteome suggests enhanced inflammation and lung damage pathways.

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BAL from AAT-KO ferrets has reduced NE inhibitory capacity, and secreted...
(A) Western blot of BAL from AAT-KO (dash) and matched control ferrets (plus); arrowhead indicates AAT band in plasma as positive and negative controls. Ponceau-stained blot as loading control. (B) NE inhibitory capacity of increasing volumes of BAL from AAT-KO and matched control ferrets (n = 5 pairs; P value by mixed effects model, P = 0.005). (C) Quantification of NE inhibitory capacity by calculating the AUC in B (n = 5 pairs; P value by Student’s t test, P = 0.0297). (D) Differential of cell types in BAL from control and AAT-KO ferrets (n = 6–7 ferrets; P value by Student’s t test, P > 0.3 for each cell type). Mac, macrophages; PMN, polymorphonuclear leukocytes. (E) Volcano plot of more than 200 BAL proteins from 7 age-matched AAT-KO and control ferrets (red circles, upregulated; blue circles, downregulated; gray circles, indeterminate or not statistically significant; n = 7 matched pairs; P value by Scaffold software t test). (F) List of selected significant disease pathways discovered using MeSH analysis of the BAL proteome. Yellow bars denote disease-associated hits that are relevant to lung diseases, and numbers in parentheses indicate number of proteins found in the pathway. (G and H) IPA performed on proteomics data where (G) canonical and (H) disease function pathways are plotted against z score and –log10(P value). In both panels, red indicates upregulated and blue downregulated pathways by z score; in H inflammation-related pathways are in orange and cancer-related pathways are in green. Graphs in B–D show mean ± SEM. *P < 0.05, **P < 0.01. FC, fold change.