Long-term expression of human adenosine deaminase in mice transplanted with retrovirus-infected hematopoietic stem cells.
Long-term expression of human adenosine deaminase in mice transplanted with retrovirus-infected hematopoietic stem cells.
复制标题
移植逆转录病毒感染的造血干细胞的小鼠体内人腺苷脱氨酶的长期表达。
DOI：
10.1073/pnas.86.22.8892
复制
发表时间：
1989
影响因子：
11.1
通讯作者：
Williams,DA
中科院分区：
文献类型：
--
作者：
Lim,B;Apperley,JF;Orkin,SH;Williams,DA
通过使用逆转录病毒载体将外源基因序列转移到造血干细胞中的长期稳定表达构成了体细胞基因治疗的相关模型。这种表达的稳定性可能取决于载体设计，包括载体内特定序列的存在或不存在，以及造血靶细胞感染的性质和效率。我们以前曾报道成功地转移人DNA编码腺苷脱氨酶（ADA）到CFU-S（集落形成单位脾）干细胞使用简化的重组逆转录病毒载体。人ADA在CFU-S衍生的脾集落中以接近内源性酶的水平表达。然而，由于缺乏有效的显性选择标记和低重组病毒滴度，未检查人ADA长期表达的稳定性。我们在这里报告的发展，一个有效的方法感染造血干细胞（HSC），而不依赖于在体外选择。100%的移植有通过该方案感染的HSC的小鼠的外周血样品在移植后30天表现出人ADA的表达。一些小鼠（6/13）在完全造血重建（4个月）后继续在所有谱系中表达人ADA。使用重组逆转录病毒载体，有效地转移人ADA cDNA到HSC中，导致稳定表达的功能性ADA在重建的小鼠，提供了一个实验框架，为未来的发展的方法，体细胞基因治疗。
Long-term stable expression of foreign genetic sequences transferred into hematopoietic stem cells by using retroviral vectors constitutes a relevant model for somatic gene therapy. Such stability of expression may depend on vector design, including the presence or absence of specific sequences within the vector, in combination with the nature and efficiency of infection of the hematopoietic target cells. We have previously reported successful transfer of human DNA encoding adenosine deaminase (ADA) into CFU-S (colony-forming unit-spleen) stem cells using simplified recombinant retroviral vectors. Human ADA was expressed in CFU-S-derived spleen colonies at levels near to endogenous enzyme. However, because of the lack of an efficient dominant selectable marker and low recombinant viral titers, stability of long-term expression of human ADA was not examined. We report here the development of an efficient method of infection of hematopoietic stem cells (HSC) without reliance on in vitro selection. Peripheral blood samples of 100% of mice transplanted with HSC infected by this protocol exhibit expression of human ADA 30 days after transplantation. Some mice (6 of 13) continue to express human ADA in all lineages after complete hematopoietic reconstitution (4 months). The use of recombinant retroviral vectors that efficiently transfer human ADA cDNA into HSC leading to stable expression of functional ADA in reconstituted mice, provides an experimental framework for future development of approaches to somatic gene therapy.
DOI：
--
发表时间：
1961
期刊：
影响因子：
--
作者：
Francis Martin
通讯作者：
Francis Martin
人腺苷脱氨酶在小鼠造血细胞中的表达
影响因子：
5.3
作者：
J. Belmont;G. MacGregor;K. Wager;F. A. Fletcher;K. A. Moore;D. Hawkins;D. Villalon;S. Chang;C. Caskey
通讯作者：
C. Caskey
逆转录病毒转移后人腺苷脱氨酶在小鼠造血祖细胞中的表达
影响因子：
64.8
作者：
J. Belmont;J. Henkel;Stephen M. W. Chang;K. Wager;R. Kellems;J. Dick;M. Magli;R. A. Phillips;A. Bernstein;C. Caskey
通讯作者：
C. Caskey