In vivo selection of gene modified hepatocytes

今天，许多其他很有前途的肝脏导向基因和细胞新策略的临床应用 治疗因其固有的数量低效而受阻。在这里，我们建议使用体内选择来 克服这些问题。我们将开发临床适用的方法来使用小分子药物来选择 体内的转基因肝细胞，从而跨过永久 治疗效果。我们已经确定发烧药物对乙酰氨基酚(APAP)是治疗 这种方法。 设计了三个特定的目标来充分验证这些用于体内治疗选择的新方法 修饰的肝细胞。 在目标1中，我们将开发可由APAP和AIM选择的集成rAAV载体(通用载体 2，将产生有条件选择的慢病毒载体。两者都将在代谢性疾病中得到验证 模特们。在目标3中，我们将把目标1和目标2的结果应用于人肝细胞。遗传选育 改良的人肝细胞将在肝嵌合小鼠中进行。

Today the clinical application of many otherwise promising new strategies for liver directed gene and cell therapy is hampered by their inherent quantitative inefficiency. Here we propose to use in vivo selection to overcome these problems. We will develop clinically applicable ways to use small molecule drugs to select genetically modified hepatocytes in vivo and thereby cross the quantitative threshold required for permanent therapeutic benefit. We have identified the fever medication acetaminophen (APAP) as excellent candidate for this approach. Three specific aims are designed to fully validate these novel method for in vivo selection of therapeutically modified hepatocytes. In Aim 1, we will develop integrating rAAV vectors (Generide vectors) that can be selected by APAP and in aim 2,conditionally selectable lentiviral vectors will be generated. Both will be validated in metabolic disease models. In Aim 3, we will apply the results of Aims 1&2 to human hepatocytes. Selection of genetically modified human hepatocytes will be carried out in liver chimeric mice.