Molecular Core

在许多方面，遗传性疾病，如囊性纤维化(CF)，是分子治疗的理想对象。然而，一些障碍，包括低效的传递/摄取和目前可用的载体的潜在毒性，仍然阻碍了临床的广泛应用。在MTCC的背景下，分子核心将以两种方式服务于北卡罗来纳大学-CH的基因转移社区。首先，它将提供新开发的小鼠模型，该模型的呼吸道模仿充满粘液的人类CF肺。这些小鼠是通过对小鼠上皮钠通道的β亚基进行呼吸道特异性过表达而发展起来的，它们表现出许多与人类CFs相同的特征，包括对钠的高吸收，纤毛周围液体减少，粘液纤毛清除减少，以及粘液积累和堵塞。因此，在CF样疾病的背景下，它们是评估递送方法和媒介的极佳模型。这个 分子核心将在不同的背景下培育和维护这些小鼠，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这一模型。其次，分子核心将提供毒物基因组学服务。这将通过利用Affymetrix基因芯片阵列进行RNA表达分析来实现。微阵列将被用来识别不同载体处理后细胞内基因表达的变化。这些数据将用于评估载体在临床环境中的潜在毒性。具体地说，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 实时定量聚合酶链式反应(罗氏LightCycler仪器和专业技术)。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实验的数据进行比较。这一机制应有助于对新的载体修饰进行有效评价，因为它们与潜在的安全性/毒性问题有关。通过为CF呼吸道疾病提供相关的模型和评估新载体的潜在毒性的有效机制，分子核心的努力应该会减少实现对CF等疾病的分子治疗的时间。

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.