Lesniak Maciej S, Kelleher Erin, Pardoll Drew, Cui Yan
Section of Neurosurgery, The University of Chicago Pritzker School of Medicine, USA.
Neurol Res. 2005 Dec;27(8):820-6. doi: 10.1179/016164105X49454.
Hematopoietic stem cells (HSC) have been previously used as vectors for gene therapy of systemic disease. The effectiveness of HSC-mediated gene therapy largely depends on efficient gene delivery into long-term repopulating progenitors and targeted transgene expression in an appropriate progeny of the transduced pluripotent HSCs. In the present study, we examined the feasibility of using HSC transduced with self-inactivating (SIN) lentiviral vectors for the delivery of gene therapy to the central nervous system (CNS).
We constructed two SIN lentiviral vectors, EF.GFP and DR.GFP, to express the green fluorescent protein (GFP) gene controlled solely by the promoter of either a housekeeping gene EF-1alpha or the human HLA-DRalpha gene, which is selectively expressed in antigen-presenting cells.
We demonstrated that both vectors efficiently transduced human pluripotent CD34+ cells capable of engrafting NOD/SCID mice. Only the DR.GFP vector mediated transgene expression in the murine CNS containing human HLA-DR+ cells. These cells express surface markers characteristic of resident CNS microglia. Furthermore, human dendritic cells derived from transduced and engrafted human cells potently stimulated allogeneic T cell proliferation.
The present study demonstrated successful targeting of transgene expression to CNS microglia after stable gene transduction of pluripotent HSC.
造血干细胞（HSC）先前已被用作全身性疾病基因治疗的载体。HSC介导的基因治疗的有效性很大程度上取决于将基因有效递送至长期重建造血祖细胞，并在转导的多能HSC的适当子代中实现靶向转基因表达。在本研究中，我们研究了使用经自我失活（SIN）慢病毒载体转导的HSC向中枢神经系统（CNS）进行基因治疗的可行性。
我们构建了两种SIN慢病毒载体，EF.GFP和DR.GFP，以表达仅由管家基因EF-1α或人HLA-DRα基因的启动子控制的绿色荧光蛋白（GFP）基因，后者在抗原呈递细胞中选择性表达。
我们证明这两种载体均能有效地转导能够植入NOD/SCID小鼠的人多能CD34+细胞。只有DR.GFP载体介导了在含有人类HLA-DR+细胞的小鼠中枢神经系统中的转基因表达。这些细胞表达驻留中枢神经系统小胶质细胞的特征性表面标志物。此外，源自转导和植入的人类细胞的人树突状细胞有力地刺激了同种异体T细胞增殖。
本研究证明在多能HSC稳定基因转导后，转基因表达成功靶向中枢神经系统小胶质细胞。
