GENE THERAPY IN HEMATOPOIETIC CELLS

这项研究的目标是应用基因转移技术。 用于治疗影响人类造血系统的疾病。 而小鼠造血系统中的基因转移率可以接近 100%，在大型动物和人类中的基因转移率 低得令人失望。我们将探索提高基因转导水平的方法 长期再生造血细胞(干细胞)的比率 犬类模型。我们将研究造血药的最佳暴露时间 干细胞到逆转录病毒载体，干细胞的最佳来源，以及 毒性较低的调理方案或不反复输液的调理 转导的造血干细胞。新载体与逆转录病毒 包装细胞株将被产生并测试改进的基因 传递和表达。犬α-L艾杜糖醛酸酶缺乏症的遗传 疾病模型将被用来测试我们纠正疾病的能力 影响狗的造血细胞。我们将继续我们目前的 转导G-CSF治疗高雪病的临床方案 动员外周血造血细胞。这件事的重点是 实验是考察载体序列的持久性和表达 在移植后。在犬类模型中发展的技术 改进的基因转移和表达将为 人类基因治疗实验的改进。最后，我们有 在检测人类造血细胞时发现了一种新的逆转录病毒 用逆转录病毒载体转导后没有辅助病毒。 新病毒似乎来自人类，我们将进行 以确定其起源的实验。如果这真的是一个 人类病毒，我们将探索病毒对正在进行的基因的影响 逆转录病毒载体的治疗及其在人类中的可能作用 疾病。

The goal of the proposed research is to apply techniques of gene transfer to the treatment of diseases affecting the hematopoietic system in humans. While gene transfer rates in the mouse hematopoietic system can approach 100%, gene transfer rates in large animals and humans have been disappointingly low. We will explore methods to improve gene transduction rates in long-term repopulating hematopoietic cells (stem cells) in the canine model. We will study the optimal exposure time of hematopoietic stem cells to retrovirus vectors, the optimal source of stem cells, and less toxic conditioning regimens or no conditioning with repeated infusions of transduced hematopoietic stem cells. New vectors and retrovirus packaging cell lines will be generated and tested for improved gene transfer and expression. The canine alpha-L-iduronidase-deficient genetic disease model will be used to test our ability to correct a disease that affects hematopoietic cells in the dog. We will continue our current clinical protocol to treat Gaucher disease by transduction of G-CSF- mobilized peripheral blood hematopoietic cells. The focus of this experiment is to examine the persistence and expression of vector sequences following transplantation. Techniques developed in the canine model for improved gene transfer and expression will provide the basis for improvements in human gene therapy experiments. Finally, we have discovered a new retrovirus during testing of human hematopoietic cells for the absence of helper virus following transduction with retroviral vectors. The new virus appears to be of human origin, and we will perform experiments to definitively establish its origin. If indeed this is a human virus, we will explore implications of the virus for ongoing gene therapy involving retrovirus vectors, and its possible role in human disease.