In Vivo Hematopoietic Stem Cell Gene Therapy of Beta-Thalassemia and Sickle Cell Disease

摘要： 造血干细胞(HSC)基因治疗可为多种血液疾病提供根治疗法。 传统的方法是基于体外HSC基因转移，并取得了令人鼓舞的结果。 然而，高昂的成本和副作用限制了患者在体外进行HSC基因治疗的可及性。我们有 利用HSC动员和静脉病毒载体建立体内HSC转导方法 注射。这种方法的突出特点是成本相对较低，技术上也很简单。它可以作为以下形式提供 门诊治疗。我们已经在几个小鼠疾病模型中证明了它的安全性和有效性， 包括β地中海贫血、镰刀细胞性贫血和血友病A，以及最近在恒河猴身上发现的。使用 更多的基因治疗产品即将问世，体内HSC转导的应用可以推断基因 为更大的患者群体提供治疗。体内HSC基因治疗的进一步改进 临床应用包括更有效的动员方案，完全消除先天反应 静脉注射载体，更高级的病毒衣壳修饰，绕过先前存在的抗载体 免疫，改进的体内选择方案，以及基因转移载体纯化的新方法。 在这一应用中，我们将陆续测试假说，以改进体内HSC基因治疗方法 小鼠地中海贫血和镰刀细胞病模型。然后，我们将验证最佳组合 人类灵长类的改进。安全性、有效性、便携性和低成本是 每个技术单元的设计/优化。

Abstract: Hematopoietic stem cell (HSC) gene therapy could provide a curative treatment for a number of blood diseases. The conventional approach is based on ex vivo HSC gene transfer and has achieved encouraging results. However, the high cost and side effects limit the patient accessibility of ex vivo HSC gene therapy. We have developed an in vivo HSC transduction approach involving HSC mobilization and intravenous viral vector injection. The approach is highlighted by its relatively low cost and technical simplicity. It could be provided as an outpatient treatment. We have demonstrated its safety and efficacy in several murine disease models, including β-thalassemia, Sickle Cell Anemia, and hemophilia A, and more recently, in rhesus macaques. With more gene therapy products on the horizon, the application of in vivo HSC transduction could extrapolate genetic treatments to a larger patient population. Further improvements of in vivo HSC gene therapy on the road to clinical application include more effective mobilization protocols, complete elimination of innate responses upon intravenous vector injection, more advanced virus capsid modifications that circumvent pre-existing anti-vector immunity, improved in vivo selection regimens, as well as new methods for purification of gene transfer vectors. In this application, we will successively test hypotheses to improve in vivo HSC gene therapy approaches for thalassemia and Sickle Cell Disease in mouse models. We will then validate the best combination of improvements in hon-human primates. Safety, efficacy, portability, and low costs are the major driving forces in the design/optimization of each technological unit.