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，仍存在一些障碍，包括目前可用载体的低效递送/摄取和潜在毒性，以阻止广泛的临床使用。MTCC背景下的分子核心将以两种方式为UNC-CH的基因转移社区服务。首先，它将提供新开发的小鼠模型，其气道模拟充满粘液的人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产生的，表现出许多与人CF相同的特征，包括Na+吸收过度、纤毛周液减少、粘液纤毛清除减少以及粘液积聚和堵塞。因此，它们作为一个很好的模型来评估CF样疾病的背景下的交付方法和载体。的 分子核心将在各种背景下繁殖和维持这些小鼠，将为研究人员提供实验动物，并将通过修改转基因品系中使用的启动子元件来扩展该模型。其次，分子核心将提供毒理基因组学服务。这将通过利用Affyssin基因芯片阵列进行RNA表达分析来完成。微阵列将用于鉴定用各种载体处理后细胞内的基因表达变化。该数据将用于评价载体在临床环境中的潜在毒性。具体而言，分子核心将为研究人员提供实验设计，RNA分离服务，进行实验所需的基因芯片阵列，数据分析/解释服务以及通过 实时定量PCR（Roche LightCycler仪器和专业知识）。此外，分子核心将维护一个数据库，使新的实验能够有效地与过去的实验数据进行比较。该机制应有助于有效评价新的载体修饰，因为它们与潜在的安全性/毒性问题有关。通过提供CF气道疾病的相关模型和评估新载体潜在毒性的有效机制，分子核心的努力应该减少实现CF等疾病分子治疗的时间。

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.