Project 3 Title: Gene Therapy for Lung Disease in a Ferret Model of Cystic Fibrosis Project Summary The development of gene therapies to the CF lung has met with many challenges over the past two decades. The greatest of these has been a lack of animal models that reproduce the human CF lung phenotype, which has been solved by the creation of CF models in ferret and pig. These models will provide insights into the basic cell biology of the disease and facilitate the development of effective treatments, including gene therapy. This proposed project will focus on evaluating two gene therapy approaches in the CF ferret model and on elucidating key aspects of airway cell biology that will direct choices of the gene therapy approaches used. CF ferrets have two lung infection phenotypes that will be useful in developing therapies for CF?rapid, lethal neonatal infections during the first 1-3 weeks of life; and slower, chronic bacterial infections thereafter, typically leading to death by 3-5 months of life. We will use these robust lung phenotypes to develop lentivirus- and recombinant adeno-associated virus (rAAV)-based gene therapies for CF lung disease. The preliminary data presented, and vectors chosen for study, in this project are the product of the first two years of an R01 award that will transition into this PPG. Important goals of this project include: 1) the development and analysis of miRNA detargeting approaches to regulate cellular expression patterns of a lentiviral CFTR transgene and limit the phenotypic consequences of ectopic CFTR expression in multi-potent airway progenitor cells, 2) the generation of rAAV-CFTR vectors that are capable of reversing lung disease in the CF ferret model and that are also compatible with both the packaging limitations of the rAAV genome and cellular expression patterns required for efficient complementation of CFTR function, and 3) identification of the cell types in the lung that must be targeted for effective complementation of CF lung disease. The third goal of the proposal draws on the unique ability of the Engelhardt laboratory to rapidly generate transgenic ferrets that express recombinant fCFTR in biologically relevant cell types within the lung. Thus, this proposal uses novel viral targeting methods to address cellular mechanisms relevant to improving gene therapies to the airway, while also tackling difficult cell biology questions about the pathogenesis and treatment of CF lung disease using gene therapy. This research is expected to significantly enhance the field's ability to develop effective therapies for CF lung disease.
(Project 3) Cystic Fibrosis (CF) is the most common life-threatening autosomal-recessive condition among Caucasians, with clinical care for patients in the U.S. alone totaling over $450 million annually. Despite the fact that the gene defect responsible for CF was discovered over 20 years ago, a cure for CF has not yet emerged?due in large part to a lack of appropriate animal models in which to study the disease process and to test therapies. The proposed research will utilize new animal models of CF to identify the cell types within the CF lung that must be corrected to prevent lethal lung infections and develop viral vectors for effective gene therapy to the CF lung.
|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|
|O'Malley, Yunxia; Rotti, Pavana G; Thornell, Ian M et al. (2018) Development of a polarized pancreatic ductular cell epithelium for physiological studies. J Appl Physiol (1985) 125:97-106|
|Rosen, Bradley H; Chanson, Marc; Gawenis, Lara R et al. (2018) Animal and model systems for studying cystic fibrosis. J Cyst Fibros 17:S28-S34|
|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|
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