Molecular Core

在许多方面，遗传性疾病，如囊性纤维化（CF），是分子治疗的理想候选者。然而，一些障碍，包括低效率的递送/摄取和目前可用的媒介的潜在毒性，仍然阻碍临床广泛使用。MTCC背景下的Molecular Core将以两种方式为UNC-CH的基因转移社区服务。首先，它将为新开发的小鼠模型提供途径，该模型的气道模拟充满粘液的人类CF肺。这些小鼠是通过小鼠上皮钠通道β亚基的气道特异性过表达而发育的，表现出许多与人类CF相同的特征，包括Na+的高吸收，睫状体周围液体减少，粘膜纤毛清除减少，粘液积聚和堵塞。因此，它们可作为评估传染性囊状病毒样疾病的递送方法和媒介的极好模型。这个

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.