KIDNEY SPECIFIC, LIPOSOME/POLYCATION MEDIATED THERAPY

描述（改编自申请人的摘要）： 这项建议是发展基因疗法来治疗遗传性 肾脏疾病 碳酸酐酶（CA）II缺陷小鼠是理想的 肾特异性基因治疗的模型，因为它表现为肾小管 酸中毒和基因治疗的效果可以很容易地监测。 CA II是一个 存在于肾脏和其他器官（如大脑）细胞中的酶， 骨，是重要的酸碱平衡，在细胞和整体 身体水平。 研究人员开发了一种新的方法， CA II基因，由病毒启动子驱动，进入尿空间， 使用脂质体作为载体的CA II缺陷小鼠的肾脏，并显示出 肾小管性酸中毒的短暂（3周）纠正。 在此期间， 在此期间，将测试新策略以提高疗效和持续时间 of gene基因expression表达using运用this gene基因therapy治疗model模型. 具体而言，目标 将：1）添加聚阳离子，带正电的有机聚合物 化合物，如碱性氨基酸，到脂质体递送系统中， 以提高体内转染效率。 这些聚阳离子可以 作为信号将治疗基因递送到细胞核; 2） 用CA II替换治疗基因的病毒启动子， 启动子，以产生细胞特异性的，可调节的，和长期的 小鼠基因表达; 3）评价 脂质体/聚阳离子介导的小鼠基因治疗;以及4）确定 CA II缺陷小鼠的基因治疗是否会改变 与CA II缺乏相关的变化，即，CA IV的上调，以及 消耗的闰细胞，这是负责酸分泌， 肾脏 希望拟议研究的结果将 从而导致适合于未来临床试验的最佳基因疗法， 人类的多种肾脏疾病。

DESCRIPTION (Adapted from the Applicant's Abstract): The ultimate goal of this proposal is to develop gene therapy for the treatment of hereditary kidney diseases. The carbonic anhydrase (CA) II deficient mouse is an ideal model for kidney-specific gene therapy since it manifests renal tubular acidosis and effects of gene therapy can be easily monitored. CA II is an enzyme present in cells of the kidney and other organs such as the brain and bones, and is important for acid-base homeostasis at both cellular and whole body levels. The investigators have developed a novel approach by infusing the CA II gene, driven by a viral promoter, into the urinary space of the kidney of the CA II deficient mice using liposomes as carriers, and shown a transient (3 week) correction of renal tubular acidosis. During this grant period, new strategies will be tested to improve the efficacy and duration of gene expression using this gene therapy model. Specifically, the goals will be: 1) to add polycations, polymers of positive-charged organic compounds such as basic amino acids, to the liposome delivery system in order to enhance the transfection efficiency in vivo. These polycations may serve as signals for the therapeutic gene to be delivered to the nucleus; 2) to replace the viral promoter of the therapeutic gene with the CA II promoter in order to produce a cell-specific, regulatable, and long-term gene expression in mice; 3) to evaluate the safety of the liposome/polycation-mediated gene therapy in mice; and 4) to determine whether gene therapy in CA II deficient mice will convert the phenotypical changes associated with CA II deficiency, i.e., up-regulation of CA IV, and depletion of intercalated cells, which are responsible for acid secretion in the kidney. It is hoped that results obtained from the proposed study will lead to an optimum gene therapy suitable for future clinical trials on a broad range of renal diseases in humans.