Cystic fibrosis (CF), the most common lethal genetic disease in the Caucasian population, results from mutations in the CFTR gene and affects the epithelia of multiple organs including the lung and gastrointestinal tract. Multiple research strategies for treatment of CF are currently being explored. Translating CF therapeutic strategies from basic research to clinical studies requires the assessment of drug candidates in physiologically relevant assays that require specific expertise. To support urgently required translational CF research, the Molecular/Functional Measurement Core (Core B) of the University of North Carolina CF Research and Translation Core Center will pursue two Specific Aims:
Specific Aim 1 will focus on in vitro evaluation of multi-organ CF disease pathophysiology and pre-clinical therapeutics candidates. We will provide in vitro analyses of CF primary cultured, stem-cell derived, and freshly biopsied epithelia (GI and airway) by Ussing chamber ion transport, biochemical measurements, and molecular studies to quantitate CFTR expression/function/maturation, organoid swelling assays to measure CFTR function in higher throughput assays, and novel models to study inflammation, infection, hypoxia, and pharmacokinetics/pharmacodynamics analyses of designated CF therapeutic agents and endpoints.
Specific Aim 2 will provide in vivo evaluation of CF disease pathophysiology and therapeutic candidates utilizing animal models. Animal models include CF mice and CF rabbits, ?-ENaC overexpressing mice, secreted and tethered mucin-deficient mice, and gnotobiotic mice. We will provide mouse model development, colony maintenance, and genotyping of mutant mice relevant to studies of CF GI and lung disease, phenotyping of nave and challenged animal models (mouse and rabbit) by validated GI and pulmonary phenotyping panels, sample collection and preparation for microbiome analyses, histopathology, immunostaining, RNAscope in situ hybridization, and q-PCR. Outcomes will include mucus burden and clearance, infection/microbiome, inflammation, pharmacokinetics, gene expression, and ion channel-mediated function (e.g., salivary secretion, nasal potential difference). Pharmacokinetics services are available for both Aims by mass spectrometry. While certain assays and models provided by the Core focus on restoration of CFTR function in CF, most of our services have broad applications to multiple research programs seeking therapeutic benefit for CF. These services include approaches that utilize small-molecule pharmacological interventions, gene therapy, correction of mucus defects, normalization of ion transport, and inflammation/infection control. The availability of these models and assays will provide a translational bridge that will support the rapid transfer of emerging drug candidates to effective therapies for CF patients.
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