Efficient transfer of genes into murine cardiac grafts by starburst polyamidoamine dendrimers
Efficient transfer of genes into murine cardiac grafts by starburst polyamidoamine dendrimers
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DOI：
10.1089/hum.1998.9.4-553
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发表时间：
1998-03-01
影响因子：
4.2
通讯作者：
Bromberg, JS
中科院分区：
文献类型：
--
作者：
Qin, LH;Pahud, DR;Bromberg, JS
在小鼠心脏移植模型中，研究了由重复的聚酰胺氨基亚基组成的结构确定的球形大分子星状树枝状聚合物（Starburst dendrimer）对质粒介导的基因转移效率的增强作用。（β-Gal）或pCH 110-树枝状聚合物复合物，并通过X-Gal染色测定报告基因表达，用pCH 110-树枝状聚合物注射的移植物从7天到28天在肌细胞和移植物浸润细胞中表现出广泛和延长的β-Gal表达，与仅在肌细胞中表达β-Gal少于14天的裸pCH 110注射的移植物相比。作为在α-肌球蛋白重链启动子控制下编码病毒白细胞介素-10（vIL-10）的质粒，p α MHC-vIL-10能够将同种异体移植物存活从13.9 +/-0.9天延长至21.4 +/-2.3天（p < 0.005）。当树枝状聚合物G5 EDA与p α MHC-vIL-10一起使用时，少60倍的DNA导致移植物存活显著延长至38.6 +/-4.7天（p < 0.0005）。在该模型系统中，DNA的剂量、DNA与树枝状聚合物的电荷比以及树枝状聚合物的尺寸生成都被确定为延长同种异体移植物存活的关键变量。因此，使用Starburst树枝状聚合物显著增加了质粒介导的基因转移和表达的效率，在更大范围的组织中以更高的量持续更长的时间产生免疫抑制细胞因子增强了免疫抑制作用并进一步延长了移植物存活。
Starburst dendrimer, a structurally defined, spherical macromolecule composed of repeating polyamidoamino subunits, was investigated to augment plasmid-mediated gene transfer efficiency in a murine cardiac transplantation model, The grafts were directly injected with naked pCH110, a plasmid encoding beta-galactosidase (beta-Gal), or pCH110-dendrimer complex, and reporter gene expression determined by X-Gal staining, The grafts injected with pCH110-dendrimer demonstrated widespread and extended beta-Gal expression in both myocytes and the graft infiltrating cells from 7 to 28 days, compared to the grafts injected with naked pCH110 that expressed beta-Gal only in myocytes for less than 14 days. p alpha MHC-vIL-10, as plasmid encoding viral interleukin-10 (vIL-10) under the control of alpha-myosin heavy chain promoter, was able to prolong allograft survival from 13.9 +/- 0.9 days to 21.4 +/- 2.3 days (p < 0.005). When dendrimer G5EDA was used with p alpha MHC-vIL-10, 60-fold less DNA resulted in significant prolongation of graft survival to 38.6 +/- 4.7 days (p < 0.0005). The dose of DNA, the charge ratio of DNA to dendrimer, and the size generation of the dendrimers were all determined to be critical variables for prolongation of allograft survival in this model system, Thus, the use of the Starburst dendrimer dramatically increased the efficiency of plasmid-mediated gene transfer and expression, Production of immunosuppressive cytokines at higher amounts for longer periods of time in a greater expanse of tissue enhanced the immunosuppressive effect and prolonged graft survival further.