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长期的研究课题。 是对实验血液学的巨大挑战。 在这个项目中，我们专注于范可尼贫血的基因替代， Diamond Blackfan贫血这两种疾病都需要成功的基因 造血干细胞（HSC）中的插入和功能及其 后代成功地将基因转移到HSC中需要选择 由宿主转染并发挥功能的HSC。这将发生在范科尼 缺乏Fanconi基因的HSC会导致贫血，并在骨髓中死亡。在 钻石黑扇贫血，选择发生在水平的 红系祖细胞和前体细胞。我们将使用两类 逆转录病毒载体;基于莫洛尼白血病的载体和基于HIV的 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， 必须利用短孵育，优选在低温下， 为了保护HSC免受由程序诱导的毒性 自己这些新的培养条件将在 实验模型适当的载体将产生在非常高的 在我们的GMP载体实验室和大规模的浓度 人体样本将在我们的GMP细胞操作中心进行 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.