Ex vivo expansion of human HSCs with Sendai virus vector expressing HoxB4 assessed by sheep in utero transplantation
Ex vivo expansion of human HSCs with Sendai virus vector expressing HoxB4 assessed by sheep in utero transplantation
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DOI：
10.1016/j.exphem.2010.09.007
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发表时间：
2011-01-01
影响因子：
2.6
通讯作者：
Hanazono, Yutaka
中科院分区：
文献类型：
--
作者：
Abe, Tomoyuki;Masuda, Shigeo;Hanazono, Yutaka
目的同源框B4（homeobox B4，HoxB 4）基因可促进造血干细胞（hematopoietic stem cells，HSC）的增殖。然而，逆转录病毒介导的HoxB 4基因在大型动物中的频繁发生导致白血病的发生。（SeV/Delta P），这使得载体和转基因能够在转导后不久被清除。用SeV/Δ P/HoxB 4转导人脐带血CD 34（+）细胞（HSC）以进行体外扩增。HoxB 4载体并移植到胎羊腹腔内。结果转染HoxB 4基因的HSCs移植后，移植后的造血干细胞存活时间较对照组延长。（长达20个月）在绵羊中繁殖，与对照未处理组相比，在HoxB 4组的骨髓中更频繁地检测到人造血祖细胞（p < 0 0 5）SeV/Delta P/HoxB 4载体对人HSCs的扩增效果与以往报道的HoxB 4逆转录病毒载体相当。SeV/Delta P/HoxB 4载体和转基因从受体羊体内清除，移植后2 0个月未检测到白血病。Delta P载体将适合于在人CD 34（+）细胞中瞬时表达HoxB 4。此外，SeV/Delta P载体不涉及转基因相关和插入性白血病发生，并且应该比逆转录病毒载体更安全（C）2011 ISFH Society for Hematology and Stem Cell Published by Elsevier Inc
Objective The homeobox B4 (HoxB4) gene promotes expansion of hematopoietic stem cells (HSCs) However, frequent development of leukemia in large animals duo to retrovirally transduced HoxB4 gene has been reported To prevent tumorigenesis, we developed a nonintegrating and nonreplicating Sendai virus vector that did not contain the phosphoprotein gene (SeV/Delta P), which enabled clearance of the vector and transgene shortly after transduction We tested the SeV/Delta P vector expressing the HoxB4 gene (SeV/Delta P/HoxB4) for the ex vivo expansion of human cord blood CD34(+) cells (HSCs) using a sheep in utero transplantation assayMaterials and Methods Human HSCs were ex vivo expanded by transduction with SeV/Delta P/HoxB4 vector and transplanted into the abdominal cavity of fetal sheep The. engraftment of human HSCs in the lambs was quantitatively evaluated by hematopoietic colony forming unit assaysResults After transplantation, the HoxB4-transduced HSCs contributed to longer period (up to 20 months) repopulation in sheep, and human hematopoietic progenitors were detected more frequently in the bone marrow of the HoxB4 group as compared with the control untreated group (p < 0 05) The expansion of human HSCs with the SeV/Delta P/HoxB4 vector was comparable with previously reported retroviral vectors expressing HoxB4 The SeV/Delta P/HoxB4 vector and the transgene were cleared from the recipient sheep and leukemia was not detected at 20 months post transplantationConclusions The SeV/Delta P vector would be suitable for transient expression of HoxB4 in human CD34(+) cells In addition, the SeV/Delta P vector is free of concern about transgene-related and insertional leukemogenesis and should be safer than retroviral vectors (C) 2011 ISFH Society for Hematology and Stem Cells Published by Elsevier Inc