ROLE OF CPG MOTIFS IN PLASMID DNA AS A BARRIER TO GENE TRANSFER

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。囊性纤维化是一种常染色体隐性遗传性疾病，其病因缺陷、功能障碍或丢失的c1通道囊性纤维化跨膜调节因子(CFTR)已被确定。这种疾病的基因治疗很有吸引力，因为最常导致死亡的受影响器官(肺)可以进行基因治疗。尽管在基因传递方面存在一些效率低下的问题，但脂质体载体介导的基因转移在纠正体内人类CFTR缺陷方面有很大的潜力。在基因治疗试验中，肺泡巨噬细胞可能是受试者的毒性来源。肺泡巨噬细胞是呼吸道中数量最多的炎性细胞。作为抗原提呈细胞和吞噬细胞，它们是肺部固有防御系统的核心参与者，并已进化为防止吸入细菌或其他病原体的感染。这项研究正在调查载体如何用于基因治疗，以及它们如何与这些灌洗液细胞或灌洗液细胞的产物相互作用，以削弱基因治疗。这将使这些研究人员能够在体外研究将肺泡巨噬细胞与其他细胞的作用联系起来的方法和基因治疗。他们假设，基因传递，即使用质粒载体的脂质体介导的基因转移，在体外具有纠正人类CFTR缺陷的巨大潜力。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Cystic fibrosis is an autosomal recessive genetic disorder in which the pathogenetic defect, dysfunction or loss of the c1-channel Cystic Fibrosis Transmembrane Regulator (CFTR) has been identified. Gene therapy for this disease is attractive because the affected organ most often responsible for death (the lung) is accessible for gene therapy. Despite some inefficiency in gene delivery, liposome-mediated gene transfer using plasmid vectors has significant potential for correction of the CFTR defect in humans in vivo. Alveolar macrophages can be a source of toxicity to the subject in gene therapy trials. Alveolar macrophages are the most numerous inflammatory cells in the airway. As antigen presenting as well as phagocytic cells, they are a central player in the innate defense system of the lung and have evolved to prevent infection from inhaled bacteria or other pathogens. This study is investigating how vectors are used in gene therapy and how they interact with these lavage cells or products of the lavage cells to impair gene therapy. This will allow these investigators to study in vitro the method of relating action of alveolar macrophages with other cells and gene therapy. They hypothesize that gene delivery, liposome-mediated gene transfer using plasmid vectors, has significant potential for correction of the CFTR defect in humans in vitro.