1. ABSTRACT (PROJECT 1) Gene transfer is a potential strategy to treat CF airways disease, yet we still lack answers to important questions to any gene transfer approach. What percentage of CF airway epithelial cells must be targeted and at what expression level is needed to correct relevant ion transport defects? What are the key CF host defense defects? Can we rescue those defects? Will targeting airway surface epithelia in vivo rescue host defense? Clinical CF lung disease? Our program made advances in three areas that allow us to answer these questions. First, we developed a model of CF in pigs. Within months of birth, they develop features of CF lung disease. Second, we discovered that loss of CFTR impairs two key respiratory host defense mechanisms, mucociliary transport (MCT) and antibacterial activity in airway surface liquid (ASL). Impaired CFTR-mediated HCO3- secretion that results in abnormally acidic ASL causes both of these defects. Third, we developed viral vectors: AAV evolved to infect airway epithelia, and GP64-pseudotyped FIV that can deliver CFTR cDNA to airway epithelia. Our preliminary data indicate that these vectors generate airway epithelial HCO3- secretion and correct the acidic ASL pH. Now is the time to answer questions that have nagged the field since we first began thinking about gene therapy. Here we propose three specific aims that will inform strategies for CF gene transfer.
Aim 1. What factors determine whether gene transfer to pig and human CF airway epithelia in vitro rescues anion secretion and host defense properties? We will investigate the relationships between transduction and expression data (the % of cells expressing CFTR, the level of CFTR expression) and the physiological consequences.
Aim 2. Will CFTR gene transfer to CF airway surface epithelia correct ASL pH, bacterial killing, and MCT defects in vivo? Aim 3. Will gene transfer to airway surface epithelia prevent the airway inflammation, infection, and remodeling associated with early CF lung disease? These data may help us understand the relative importance of the CF host defense defects, will suggest whether targeting surface epithelia has clinical benefit, and will guide future gene transfer approaches.
(PROJECT 1) Our recent discoveries indicate that CF lung disease is caused by defects in two key host defenses of the lungs: mucociliary transport (MCT) and antibacterial activity in airway surface liquid (ASL). Both of these defects are related to impaired CFTR mediated HCO3- secretion, resulting in abnormally acidic ASL and submucosal gland secretions. The series of experiments proposed will help us understand the relative importance of these CF host defense defects, will suggest whether targeting surface epithelia has clinical benefit, and will guide future gene transfer approaches.
| Meyerholz, David K; Sieren, Jessica C; Beck, Amanda P et al. (2018) Approaches to Evaluate Lung Inflammation in Translational Research. Vet Pathol 55:42-52 |
| Rosen, Bradley H; Evans, T Idil Apak; Moll, Shashanna R et al. (2018) Infection Is Not Required for Mucoinflammatory Lung Disease in CFTR-Knockout Ferrets. Am J Respir Crit Care Med 197:1308-1318 |
| Mao, Suifang; Shah, Alok S; Moninger, Thomas O et al. (2018) Motile cilia of human airway epithelia contain hedgehog signaling components that mediate noncanonical hedgehog signaling. Proc Natl Acad Sci U S A 115:1370-1375 |
| Montoro, Daniel T; Haber, Adam L; Biton, Moshe et al. (2018) A revised airway epithelial hierarchy includes CFTR-expressing ionocytes. Nature 560:319-324 |
| Lynch, Thomas J; Anderson, Preston J; Rotti, Pavana G et al. (2018) Submucosal Gland Myoepithelial Cells Are Reserve Stem Cells That Can Regenerate Mouse Tracheal Epithelium. Cell Stem Cell 22:653-667.e5 |
| Meyerholz, David K; Stoltz, David A; Gansemer, Nick D et al. (2018) Lack of cystic fibrosis transmembrane conductance regulator disrupts fetal airway development in pigs. Lab Invest 98:825-838 |
| Gray, Robert D; Hardisty, Gareth; Regan, Kate H et al. (2018) Delayed neutrophil apoptosis enhances NET formation in cystic fibrosis. Thorax 73:134-144 |
| Thornell, Ian M; Li, Xiaopeng; Tang, Xiao Xiao et al. (2018) Nominal carbonic anhydrase activity minimizes airway-surface liquid pH changes during breathing. Physiol Rep 6: |
| Reznikov, Leah R; Meyerholz, David K; Abou Alaiwa, Mahmoud et al. (2018) The vagal ganglia transcriptome identifies candidate therapeutics for airway hyperreactivity. Am J Physiol Lung Cell Mol Physiol 315:L133-L148 |
| Meyerholz, David K; Beck, Amanda P; Goeken, J Adam et al. (2018) Glycogen depletion can increase the specificity of mucin detection in airway tissues. BMC Res Notes 11:763 |
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