The Shraga Segal Department of Microbiology, Immunology, and Genetics, Faculty of Health Sciences, National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev, Beer Sheva, 84105, Israel.
Sci Rep. 2020 Feb 4;10(1):1812. doi: 10.1038/s41598-020-58724-x.
Haematopoietic stem cells (HSCs) have the potential for lifetime production of blood and immune cells. The introduction of transgenes into HSCs is important for basic research, as well as for multiple clinical applications, because HSC transplantation is an already established procedure. Recently, a major advancement has been reported in the use of cyclosporine H (CsH), which can significantly enhance the lentivirus (LV) transduction of human haematopoietic stem and progenitor cells (HSPCs). In this study, we employed CsH for LV transduction of murine HSCs and defined haematopoietic progenitors, confirming previous findings in more specific subsets of primitive haematopoietic cells. Our data confirm increased efficiencies, in agreement with the published data. We further experimented with the transduction with the simultaneous use of several vectors. The use of CsH yielded an even more robust increase in rates of multi-vector infection than the increase for a single-vector. CsH was reported to reduce the innate resistance mechanism against LV infection. We indeed found that additional pretreatment could increase the efficiency of transduction, in agreement with the originally reported results. Our data also suggest that CsH does not reduce the efficiency of transplantation into immune-competent hosts or the differentiation of HSCs while enhancing stable long-term expression in vivo. This new additive will surely help many studies in animal models and might be very useful for the development of novel HSC gene therapy approaches.
造血干细胞 (HSCs) 具有终生产生血液和免疫细胞的潜力。将转基因引入 HSCs 对于基础研究以及多种临床应用都很重要,因为 HSC 移植已经是一种既定的程序。最近,有报道称环孢素 H (CsH) 的使用有了重大进展,它可以显著提高慢病毒 (LV) 对人造血干细胞和祖细胞 (HSPCs) 的转导效率。在这项研究中,我们使用 CsH 对小鼠 HSCs 和造血祖细胞进行 LV 转导,证实了之前在更原始的造血细胞亚群中发现的结果。我们的数据证实了与已发表数据一致的更高效率。我们进一步实验了同时使用多个载体的转导。与单个载体相比,CsH 的使用甚至使多载体感染的速率增加得更为显著。CsH 被报道可以降低针对 LV 感染的固有抵抗机制。我们确实发现,额外的预处理可以提高转导效率,这与最初报道的结果一致。我们的数据还表明,CsH 不会降低在免疫功能正常的宿主中移植的效率,也不会降低 HSCs 的分化,同时增强体内稳定的长期表达。这种新的添加剂肯定会有助于许多动物模型的研究,并且对于开发新的 HSC 基因治疗方法可能非常有用。