CORRECTION OF CONGENITAL DISEASES BY STEM CELL THERAPY

先天性骨髓疾病的基因替代治疗是一个漫长的过程 寻求治疗目标和对实验血液学的巨大挑战。 在这个项目中，我们关注范可尼贫血的基因替换和 戴蒙德·布莱克凡贫血。这两种疾病都需要成功的基因 造血干细胞(HSC)的插入和功能及其机制 后代。成功地将基因转移到HSC需要选择 由宿主转染并发挥作用的HSC。这将发生在范科尼 贫血是因为缺乏Fanconi基因的HSC会在骨髓中死亡。在……里面 钻石黑粉贫血，选择发生在 红系祖细胞和前体细胞。我们将使用两个类 逆转录病毒载体；基于莫洛尼白血病的载体和基于艾滋病毒的载体 慢病毒载体。这些将在体外和两个体内模型中进行测试； NOD SCID小鼠和恒河猴。我们已经证明了这一点 必须利用短期孵化，最好是在寒冷的温度下 以保护肝星状细胞免受手术引起的毒性 他们自己。这些新的孵化条件将在 实验模型。适当的载体将在非常高的位置产生 我们的GMP媒介实验室的浓度和规模达到了很大 人体样本将在我们的GMP细胞操作核心进行 实验室。

Gene replacement therapy of congenital bone marrow disorders is a long sought treatment goal and a great challenge to experimental hematology. In this project we focus on gene replacement for Fanconi Anemia and Diamond Blackfan Anemia. Both diseases require successful gene insertion and function in hematopoietic stem cells (HSC) and their progeny. Successful gene transfer into HSC requires selection of transfected and functioning HSC by the host. This will occur in Fanconi Anemia because HSC lacking the Fanconi gene die in the marrow. In Diamond Blackfan Anemia, the selection occurs at the level of the erythroid progenitor and precursor cells. We will use two classes of retrovirus vectors; moloney leukemia based vectors and HIV based lentivirus vectors. These will be tested in vitro and in two in vivo models; the NOD SCID mouse and in rhesus monkey. We have shown that it is imperative to utilize short incubations preferably in cold temperatures in order to protect the HSC from toxicity induced by the procedures themselves. These new incubation conditions will be tested in the experimental models. Appropriate vectors will be produced at very high concentrations in our GMP vector laboratory and the scale up to large human samples will be carried out in our GMP cell manipulation core laboratory.