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IDURONIDASE GENE TRANSFER INTO HEMATOPOIETIC CELLS

艾杜糖醛酸酶基因转移至造血细胞

基本信息

批准号：
6915315
负责人：
FRIEDRICH G SCHUENING
金额：
$ 7.55万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-09-30 至 2005-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6915315
关键词：
O glycosidase artificial immunosuppression autologous transplantation bone marrow transplantation dogs gene therapy genetic transduction hematopoietic stem cells human immunodeficiency virus 1 immunity mucopolysaccharidosis type I murine leukemia virus technology /technique development transfection

项目摘要

Metabolic storage disease like mucopolysaccharidosis can potentially be cured by ex vivo transfer of the deficient gene into autologous hematopoietic stem cells followed by infusion of the genetically corrected cells. Transduction efficiency into hematopoietic stem cells of large animals and humans though has been too low to achieve a therapeutic effect. In this proposal, we wish to evaluate strategies with low toxicity to increase engraftment of hematopoietic stem cells transduced with Moloney murine leukemia virus (MoMLV) vectors transduced marrow cells in the absence of conditioning or infusions after partial marrow ablation with either hydroxyurea, methotrexate or busulfan are at least as effective as 200 cGy TBI, shown to be more effective in our past studies. Another hurdle towards gene therapy is the possible induction of immune responses to transduced cells and the transgene encoded proteins. We propose to characterize the immune response in normal beagles to autologous marrow cells transduced with the "foreign" human IDU gene to correlate the long-term persistence of these transduced cells with the host's immune response. We wish to develop methods to prevent or decrease this response by immunosuppressive treatment after transplantation. The most effective method will be tested in IDU-deficient dogs which will receive autologous marrow cells transduced with the "foreign" canine IDU gene followed by immunosuppressive treatment. Hematopoietic stem cells are considered to be predominantly quiescent. Gene transfer into hematopoietic stem cells by MoMLV vectors is not efficient in part due to their inability to transduce non-dividing cells. As human immunodeficiency virus type 1 (HIV-1) based vectors have been shown to infect non-dividing target cells, we wish to compare the efficiency of HIV-1 vectors containing the canine IDU gene with corresponding MoMLV vectors to transduce stem cells of normal and IDU deficient dogs. The outlined studies will address current problems of gene transfer and, if successful, will facilitate future gene therapy of Hurler syndrome and other diseases.

代谢性胆积病如粘多糖样沉积症可以通过将缺陷基因离体转移到自体造血干细胞中，然后输注遗传校正的细胞来治愈。然而，转导到大型动物和人类的造血干细胞中的效率太低，无法达到治疗效果。在该提议中，我们希望评估具有低毒性的策略，以增加用莫洛尼鼠白血病病毒（MoMLV）载体转导的造血干细胞的植入。在不存在预处理的情况下，或者在用羟基脲、甲氨蝶呤或白消安进行部分骨髓消融后进行输注的情况下，用莫洛尼鼠白血病病毒（MoMLV）载体转导的骨髓细胞至少与200 cGy TBI一样有效，在我们过去的研究中显示出更有效。基因治疗的另一个障碍是可能诱导对转导细胞和转基因编码蛋白质的免疫应答。我们建议表征正常比格犬对用“外来”人IDU基因转导的自体骨髓细胞的免疫应答，以将这些转导细胞的长期持久性与宿主的免疫应答相关联。我们希望开发方法来预防或减少这种反应的免疫抑制治疗移植后。最有效的方法将在IDU缺陷犬中进行测试，这些犬将接受用“外源”犬IDU基因转导的自体骨髓细胞，然后进行免疫抑制治疗。造血干细胞被认为主要是静止的。通过MoMLV载体将基因转移到造血干细胞中不是有效的，部分原因是它们不能使非分裂细胞增殖。由于基于人类免疫缺陷病毒1型（HIV-1）的载体已被证明感染非分裂靶细胞，我们希望比较含有犬IDU基因的HIV-1载体与相应MoMLV载体对正常和IDU缺陷犬的干细胞增殖的效率。概述的研究将解决目前的问题，基因转移，如果成功，将促进未来的基因治疗赫尔勒综合征和其他疾病。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Novel Tat-encoding bicistronic human immunodeficiency virus type 1-based gene transfer vectors for high-level transgene expression.

新型 Tat 编码双顺反子人类免疫缺陷病毒 1 型基因转移载体，用于高水平转基因表达。

DOI：
10.1128/jvi.74.14.6659-6668.2000
发表时间：
2000
期刊：
Journal of virology
影响因子：
5.4
作者：
Srinivasakumar,N;Schuening,F
通讯作者：
Schuening,F

Evaluation of Tat-encoding bicistronic human immunodeficiency virus type 1 gene transfer vectors in primary canine bone marrow mononuclear cells.

原代犬骨髓单核细胞中编码 Tat 的双顺反子人类免疫缺陷病毒 1 型基因转移载体的评价。

DOI：
10.1128/jvi.76.14.7334-7342.2002
发表时间：
2002
期刊：
Journal of virology
影响因子：
5.4
作者：
Srinivasakumar,Narasimhachar;Zaboikin,Michail;Zaboikina,Tatiana;Schuening,Friedrich
通讯作者：
Schuening,Friedrich

Cloning and expression of canine O6-methylguanine-DNA methyltransferase in target cells, using gammaretroviral and lentiviral vectors.

使用γ逆转录病毒和慢病毒载体在靶细胞中克隆和表达犬O6-甲基鸟嘌呤-DNA 甲基转移酶。

DOI：
10.1089/104303404322959533
发表时间：
2004
期刊：
Human gene therapy.
影响因子：
0
作者：
Zaboikin,Michail;Srinivasakumar,Narasimhachar;Zaboikina,Tatiana;Schuening,Friedrich
通讯作者：
Schuening,Friedrich

A lentivirus packaging system based on alternative RNA transport mechanisms to express helper and gene transfer vector RNAs and its use to study the requirement of accessory proteins for particle formation and gene delivery.

一种基于替代RNA运输机制的慢病毒包装系统，用于表达辅助RNA和基因转移载体RNA，及其用于研究颗粒形成和基因递送的辅助蛋白的需求。