Novel Gene Editing Approaches for Hemoglobinopathies

血红蛋白生成障碍是最常见的遗传性造血障碍，是一种 在全球范围内造成巨大的人类疾病和经济损失。基因编辑的最新进展 技术已经证明了直接操纵人类内源性珠蛋白基因的可能性 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，如 珠蛋白产生障碍的有效治疗方法，包括1)基因编辑珠蛋白的定义 支持高水平非镰状珠蛋白生产的基因座架构，2)维持多血统 基因编辑的HSPC的重建能力，以及3)高效植入和 基因编辑的HPSC的选育。在本申请中，我们提出了两个协调一致的具体目标，它们共同 解决这些障碍。特定目标1将利用最近发布的方法进行高效的HSPC基因编辑 定义支持高水平红系特异性珠蛋白产生的珠蛋白基因座架构(S) 启用基因编辑细胞的植入后选择。具体目标2将评估嫁接和选择 临床相关猕猴自体移植模型中基因编辑HSPC的方案。总体而言， 拟议的研究将定义优化的珠蛋白基因编辑程序以及启用IND的临床前 将作为β-珠蛋白疾病治疗性基因编辑翻译的基础的数据集 合成到人类临床试验I期。

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.