Multifunctional peptide-PEG intercalating conjugates: programmatic of gene delivery to the blood-brain barrier.
Multifunctional peptide-PEG intercalating conjugates: programmatic of gene delivery to the blood-brain barrier.
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DOI：
10.1007/s11095-010-0256-x
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发表时间：
2010-12
影响因子：
3.7
通讯作者：
Vinogradov, Serguei V.
中科院分区：
文献类型：
--
作者：
Zhang, Hongwei;Gerson, Trevor;Varney, Michelle L.;Singh, Rakesh K.;Vinogradov, Serguei V.
关键词：
通过应用多功能肽- peg -三-吖啶偶联物（pPAC）和形成可生物降解的核-壳多聚体，将基因传递到血脑屏障（BBB），提高pDNA的转染效率。在小鼠血脑屏障细胞中对ppac介导的转染进行了成分优化（bEnd.3）。通过荧光成像和组织化学评估pDNA的细胞摄取和运输以及脑积累。我们构建了抗MRP4 sirna产生载体，并通过Western blot和qPCR评估MRP4下调MRP4的效果，以及其对MRP4底物3H-AZT摄取的影响。由pDNA和pPAC组装成核壳基因传递系统（GDS），携带NLS、TAT和脑特异性BH或ApoE序列的多功能肽，以及可生物降解的pLPEI多胺。这种GDS在慢分裂的bEnd中表现出更好的细胞和细胞核积累，转染效率高出25倍。3个细胞与ExGen500相比。脑靶向pPAC的加入增强了脑毛细血管中功能性pDNA的体内积累。用包封抗MRP4 sirna产生的pDNA处理引起MRP4的短暂下调，并且在Balb/c小鼠中静脉注射后，大脑中AZT的摄取增加了230-270%。pPAC代表了GDS的新型有效成分，可用于各种基因治疗，包括血脑屏障的遗传调节。
To enhance transfection efficacy of pDNA through the application of multifunctional peptide-PEG-tris-acridine conjugates (pPAC) and the formation of biodegradable core-shell polyplexes for gene delivery to the blood-brain barrier (BBB). pPAC-mediated transfection was compositionally optimized in mouse BBB cells (bEnd.3). Cellular uptake and trafficking, and brain accumulation of pDNA was evaluated by fluorescent imaging and histochemistry. We constructed anti-MRP4 siRNA-producing vectors and evaluated the efficacy of MRP4 down-regulation of MRP4 by Western blot and qPCR, and its effect on the uptake of 3H-AZT, an MRP4 substrate. A core-shell gene delivery system (GDS) was assembled from pDNA and pPAC, carrying multifunctional peptides with NLS, TAT, and brain-specific BH, or ApoE sequences, and biodegradable pLPEI polyamine. This GDS demonstrated better cellular and nuclear accumulation, and a 25-fold higher transfection efficacy in slow-dividing bEnd.3 cells compared to ExGen500. Inclusion of brain-targeting pPAC enhanced in vivo accumulation of functional pDNA in brain capillaries. Treatment by encapsulated anti-MRP4 siRNA-producing pDNA caused transient down-regulation of MRP4, and, after intravenous injection in Balb/c mice, enhanced AZT uptake in the brain by 230–270%. The pPAC represent novel efficient components of GDS that could find various gene therapy applications, including genetic modulation of the BBB.
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影响因子：
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