In vivo selection of gene modified hepatocytes

如今，许多其他有前途的肝脏定向基因和细胞新策略的临床应用 治疗因其固有的定量低效率而受到阻碍。在这里我们建议使用体内选择 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 体内对肝细胞进行基因改造，从而跨越了永久肝细胞所需的数量阈值 治疗益处。我们已确定发烧药物对乙酰氨基酚 (APAP) 是最佳候选药物 这种方法。 设计了三个具体目标来充分验证这些用于体内选择治疗药物的新方法 修饰的肝细胞。 在目标1中，我们将开发可被APAP选择并用于目标的整合rAAV载体（Generide载体） 2、产生条件选择的慢病毒载体。两者都将在代谢疾病中得到验证 模型。在目标 3 中，我们将目标 1 和 2 的结果应用于人类肝细胞。基因选择 修饰的人类肝细胞将在肝脏嵌合小鼠中进行。

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.