Transient silencing of PTEN in human CD34(+) cells enhances their proliferative potential and ability to engraft immunodeficient mice.
Transient silencing of PTEN in human CD34(+) cells enhances their proliferative potential and ability to engraft immunodeficient mice.
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DOI：
10.1016/j.exphem.2011.10.001
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发表时间：
2012-01
影响因子：
2.6
通讯作者：
Larochelle, Andre
中科院分区：
文献类型：
--
作者：
Kim, Inho;Kim, Yoo-Jin;Metais, Jean-Yves;Dunbar, Cynthia E.;Larochelle, Andre
体外扩增造血干细胞和祖细胞（HSPC）的能力将提高广泛的移植相关疗法的成功率。PTEN（10号染色体上缺失的磷酸酶和张力蛋白同源物）被认为是鼠HSPC自我更新的调节因子，但对PTEN在人HSPC调节中的作用了解甚少。我们测试了瞬时小干扰RNA（siRNA）诱导的人CD 34+细胞中PTEN表达的抑制对其细胞周期谱、逆转录病毒转导的易感性以及其自我更新和再增殖具有IL 2受体γ（G）链缺陷的非肥胖糖尿病（NOD）/严重联合免疫缺陷病（SCID）小鼠（NSG小鼠）的能力的影响。与对照siRNA处理的CD 34+细胞相比，在PTEN siRNA处理的CD 34+细胞中证实了降低的PTEN mRNA和蛋白质水平。与对照siRNA处理的CD 34+细胞相比，瞬时沉默CD 34+细胞中的PTEN促进其进入细胞周期，并增加其体外扩增。当用逆转录病毒载体转导这些细胞时，与用对照siRNA转染的CD 34+细胞相比，用PTEN siRNA转染的大量CD 34+细胞中的转导效率显著更高。CD 34+细胞中的瞬时PTEN抑制也增加了它们在NSG小鼠中的增殖和植入潜力，并保持了它们在体内的多系分化能力。没有小鼠发生骨髓增生性疾病或白血病。与鼠HSPC的发现类似，PTEN也可促进人HSPC的静止。随着siRNA在原代CD 34+细胞中转移技术的优化，这种方法可能有助于研究HSPC自我更新的机制，并可能在基因治疗方案中找到临床应用。
The ability to expand hematopoietic stem and progenitor cells (HSPCs) in vitro will enhance the success of a wide range of transplant-related therapies. PTEN (phosphatase and tensin homologue deleted on chromosome 10) has been implicated as a regulator of murine HSPC self-renewal but little is understood about the role of PTEN in human HSPC regulation. We tested the impact of transient small interfering RNA (siRNA) - induced inhibition of PTEN expression in human CD34+ cells on their cell cycle profile, their susceptibility to retroviral transduction, and their ability to self-renew and repopulate non-obese diabetic (NOD)/severe combined immunodeficiency disease (SCID) with IL2 receptor gamma (G) chain deficiency mice (NSG mice). Reduced PTEN mRNA and protein levels were confirmed in PTEN siRNA treated CD34+ cells compared with control siRNA treated CD34+ cells. Transient silencing of PTEN in CD34+ cells promoted their entry into cell cycle, and increased their expansion in vitro compared with control siRNA treated CD34+ cells. When these cells were transduced with retroviral vectors, transduction efficiencies in the bulk CD34+ cells transfected with PTEN siRNA were significantly higher compared with CD34+ cells transfected with a control siRNA. Transient PTEN suppression in CD34+ cells also increased their proliferation and engraftment potential in NSG mice, and maintained their multilineage differentiation capacity in vivo. No mice developed myeloproliferative disorders or leukemias. Similar to findings with murine HSPC, PTEN may also promote quiescence of human HSPC. With optimization of technologies for transfer of siRNA in primary CD34+ cells, this approach may facilitate investigations into the mechanisms underlying HSPC self-renewal, and could find clinical applications in gene therapy protocols.
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影响因子：
4
作者：
Von Levetzow, G;Spanholtz, J;Giebel, B
通讯作者：
Giebel, B
DOI：
10.1002/cyto.990020302
发表时间：
1981-01-01
期刊：
CYTOMETRY
影响因子：
--
作者：
SHAPIRO, HM
通讯作者：
SHAPIRO, HM
DOI：
10.1073/pnas.0509939103
发表时间：
2006-01-03
影响因子：
11.1
作者：
Groszer, M;Erickson, R;Wu, H
通讯作者：
Wu, H
影响因子：
1.7
作者：
Klatt, Andreas R.;Klinger, Gabriele;Wielckens, Klaus
通讯作者：
Wielckens, Klaus
影响因子：
64.8
作者：
Yilmaz, Omer H.;Valdez, Riccardo;Morrison, Sean J.
通讯作者：
Morrison, Sean J.