Development of peptide-targeted lipoplexes to CXCR4-expressing rat glioma cells and rat proliferating endothelial cells
Development of peptide-targeted lipoplexes to CXCR4-expressing rat glioma cells and rat proliferating endothelial cells
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DOI：
10.1038/sj.mt.6300388
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发表时间：
2008-03-01
影响因子：
12.4
通讯作者：
Sullivan, Sean M.
中科院分区：
文献类型：
--
作者：
Driessen, Wouter H. P.;Fujii, Nobutaka;Sullivan, Sean M.
一种肽类似物，4-氟苯甲酰- rr -（L-3-（2-萘基）丙氨酸）- cyek -（l-瓜氨酸）- pyr -（l-瓜氨酸）- cr，共价连接到磷脂，用于靶向基于脂质的基因传递载体到CXCR4(+)-细胞。利用表达CXCR4的转化大鼠胶质瘤细胞系（RG2）在体外完成转染活性的表征。以增加摩尔百分比的靶脂代替辅助脂，报告基因的表达逐渐增加，在配体浓度为20摩尔%时达到控制值的2.5倍。用半胱氨酸衍生磷脂类似物或磷脂酰乙醇胺替代辅助脂质导致转染活性逐渐降低，在20 mol%时复合物完全失活。10 mol%脂肽的DNA剂量反应使有效DNA剂量相对于转染细胞的数量减少至少5倍，相对于基因表达量减少至少20倍。在动脉器官培养的背景下，研究了基因转移到大鼠内皮细胞。肠系膜动脉插管并在培养中维持4天。用血管内皮生长因子（VEGF）孵育一夜后，诱导内皮细胞表面表达CXCR4。基因转移研究表明，只有肽靶向的脂质体才能转染内皮，并且只有在CXCR4被VEGF诱导后才能转染内皮。这些结果表明，非病毒转染复合物可以靶向表达CXCR4的细胞，并且基因转移依赖于细胞表面受体的表达水平。
A peptide analog, 4-fluorobenzoyl-RR-(L-3-(2-naphthyl)alanine)-CYEK-(L-citrulline)-PYR-(L-citrulline)-CR, covalently linked to a phospholipid, was used for targeting a lipid-based gene delivery vehicle to CXCR4(+)-cells. Characterization of transfection activity was done in vitro using a transformed rat glioma cell line (RG2) that expresses CXCR4. The substitution of the targeting lipid at increasing mole percentages in the place of helper lipids yielded a progressive increase in reporter gene expression, reaching a maximum of 2.5 times the control value at 20 mol% of ligand. The substitution of helper lipids with cysteine-derivatized phospholipid analog or phosphatidylethanolamine resulted in a progressive decrease in transfection activity, with complete inactivation of the complex occurring at 20 mol%. A DNA dose-response with 10 mol% of lipopeptide reduced the effective DNA dose at least fivefold with regard to the number of transfected cells and > 20- fold with regard to the amount of gene expression. Gene transfer to rat endothelial cells was studied in the context of an arterial organ culture. Mesenteric arteries were cannulated and maintained in culture for up to 4 days. CXCR4 cell-surface expression on endothelial cells was induced after overnight incubation with vascular endothelial growth factor (VEGF). Gene transfer studies showed that only the peptide-targeted lipoplexes transfected the endothelium, and only after CXCR4 had been induced with VEGF. These results demonstrate that non-viral transfection complexes can be targeted to cells expressing CXCR4, and that gene transfer is dependent upon cell surface receptor expression levels.