In vivo selection of gene modified hepatocytes

目前，许多有前途的肝定向基因和细胞治疗新策略的临床应用， 治疗受到其固有的定量低效的阻碍。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，使用小分子药物来选择 基因修饰的肝细胞，从而越过永久性免疫所需的定量阈值。 治疗益处。我们已经确定了发热药物对乙酰氨基酚（APAP）作为 这种方法。 设计了三个具体目的以充分验证这些用于体内选择治疗性药物的新方法。 修饰肝细胞。 在目标1中，我们将开发整合rAAV载体（Generide载体），其可以通过APAP和目标中选择 2，将产生条件选择性慢病毒载体。两者都将在代谢性疾病中得到验证 模型在目标3中，我们将目标1和2的结果应用于人肝细胞。遗传选择 修饰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.