Novel Gene Editing Approaches for Hemoglobinopathies

血红蛋白生成障碍是最常见的造血遗传性障碍，并且是一种遗传性疾病。 造成地球仪巨大的人类发病率和经济损失。基因编辑的最新进展 技术已经证明了直接操纵人内源性珠蛋白基因座的潜力 造血干细胞和祖细胞。然而，基因编辑的翻译仍然存在重大障碍， 一种有效治疗珠蛋白产生障碍的方法，包括1）基因编辑珠蛋白的定义 支持高水平的非镰状球蛋白产生的基因座结构，2）维持多谱系 基因编辑的HSPC的重建能力，以及3）开发用于有效植入的方法， 选择基因编辑的HPSC。在本申请中，我们提出了两个协调的具体目标， 解决这些障碍。Specific Aim 1将利用最近发表的方法进行有效的HSPC基因编辑 以定义支持高水平红系特异性珠蛋白产生的珠蛋白基因座结构， 使基因编辑的细胞的移植后选择成为可能。具体目标2将评价植入和选择 在临床相关猕猴自体移植模型中进行基因编辑的HSPC的方案。统称 拟议的研究将定义一种优化的珠蛋白基因编辑程序，沿着IND使能的临床前 数据集，将作为β-珠蛋白疾病的治疗性基因编辑翻译的基础 在人类中进行I期临床试验。

Disorders of hemoglobin production are the most common inherited disorders of hematopoiesis, and are a cause of immense human morbidity and economic loss across the globe. Recent advances in gene-editing technology have demonstrated the potential to directly manipulate the endogenous globin locus in human hematopoietic stem and progenitor cells. However, significant barriers remain for translation of gene editing as an effective therapeutic approach to disorders of globin production, including 1) definition of gene-edited globin locus architectures that support high levels of non-sickling globin production, 2) maintenance of multi-lineage reconstitution capability of gene-edited HSPC, and 3) development of methods for efficient engraftment and selection of gene-edited HPSC. In this application, we propose two coordinated specific aims that collectively address these barriers. Specific Aim 1 will utilize recently published methods for efficient HSPC gene editing to define globin locus architecture(s) that support high levels of erythroid specific globin production and that enable post-engraftment selection of gene-edited cells. Specific Aim 2 will evaluate engraftment and selection protocols for gene-edited HSPC in a clinically relevant macaque autologous transplant model. Collectively, the proposed studies will define an optimized globin gene editing procedure along with a IND-enabling pre-clinical data set that will serve as a foundation for translation of therapeutic gene editing for disorders of β-globin synthesis to phase I clinical testing in humans.