In many ways, genetic diseases, such as cystic fibrosis (CF), are ideal candidates for molecular therapy. However, several obstacles, including inefficient delivery/uptake and the potential toxicity of presently available vectors, remain to prevent widespread clinical use. The Molecular Core in the context of the MTCC will serve the gene transfer community at UNC-CH in two ways. First, it will provide access to newly developed mouse model whose airways mimic the mucus filled human CF lung. These mice, which were developed by airway-specific overexpression of the beta subunit of the mouse epithelial sodium channel, exhibit many of the same characteristics of a human CF, including hyperabsorption of Na+, reduced periciliary liquid, decreased mucociliary clearance, and mucus accumulation and plugging. As such, they serve as an excellent model to evaluate delivery methods and vectors in the context of CF-like disease. The Molecular Core will breed and maintain these mice on a variety of backgrounds, will provide experimental animals to investigators, and will expand upon this model by modifying the promoter elements used in the transgenic lines. Secondly, the Molecular Core will provide toxicogenomic services. This will be accomplished by utilizing Affymetrix GeneChip arrays for RNA expression analysis. Microarrays will be used to identify gene expression changes within cells after treatment with various vectors. This data will be used to evaluate the potential toxicity of the vector in a clinical setting. Specifically, the Molecular Core will provide investigators with expertise on experimental design, RNA isolation services, the GeneChip arrays needed to conduct the experiment, data analysis/interpretation services, and access to validation via quantitative real-time PCR (Roche LightCycler instrumentation and expertise). In addition, the Molecular Core will maintain a database that will allow new experiments to be efficiently compared with data from past experiments. This mechanism should facilitate efficient evaluation of new vector modifications as they relate to the potential safety/toxicity issues. By providing a relevant model for CF airways disease and an efficient mechanism to evaluate the potential toxicity of new vectors, the efforts of the Molecular Core should reduce the time to achieve molecular therapy for diseases such as CF.
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