Development of viral nanoparticles for efficient intracellular delivery.
Development of viral nanoparticles for efficient intracellular delivery.
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DOI：
10.1039/c2nr30366c
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发表时间：
2012-06-07
期刊：
影响因子：
6.7
通讯作者：
Steinmetz NF
中科院分区：
文献类型：
--
作者：
Wu Z;Chen K;Yildiz I;Dirksen A;Fischer R;Dawson PE;Steinmetz NF
基于植物病毒（如豇豆花叶病毒（CPMV））的病毒纳米颗粒（VNPs）可用于广泛的生物医学应用，因为它们提供了一个强大的支架，可以通过化学偶联和基因修饰实现功能化，从而提供了一个有效的药物传递平台，可以针对特定的细胞和组织。CPMV等VNPs对细胞具有天然亲和力；然而，细胞摄取是低效的。在这里，我们展示了用高活性、特异性和紫外线可追踪的腙连接剂对CPMV表面进行化学修饰，可以实现聚精氨酸（R5）细胞穿透肽（CPPs）的生物偶联，从而克服了这些限制。所得到的CPMV-R5颗粒比天然颗粒更有效地进入人宫颈癌细胞系（HeLa）。摄取效率取决于VNP表面R5肽的密度；每CPMV显示40个R5肽的粒子（表示为CPMV - r5h）与质膜强烈相互作用，并通过能量依赖机制被吸收到细胞中，而每CPMV显示10个R5肽的粒子（表示为CPMV - r5l）只被缓慢吸收。在共定位时间过程研究中，评估了细胞内CPMV - r5与天然CPMV颗粒的命运。结果表明，CPMV - r5与CPMV的胞内定位存在差异；CPMV在很长一段时间内被困在Lamp-1阳性内溶酶体中；相比之下，30-50%的CPMV-R5颗粒从核内体转移到其他细胞囊泡或室中。我们的数据为开发基于CPMV-R5的高效给药制剂提供了基础。
Viral nanoparticles (VNPs) based on plant viruses such as Cowpea mosaic virus (CPMV) can be used for a broad range of biomedical applications because they present a robust scaffold that allows functionalization by chemical conjugation and genetic modification, thereby offering an efficient drug delivery platform that can target specific cells and tissues. VNPs such as CPMV show natural affinity to cells; however, cellular uptake is inefficient. Here we show that chemical modification of the CPMV surface with a highly reactive, specific and UV-traceable hydrazone linker allows bioconjugation of polyarginine (R5) cell penetrating peptides (CPPs), which can overcome these limitations. The resulting CPMV–R5 particles were taken up into a human cervical cancer cell line (HeLa) more efficiently than native particles. Uptake efficiency was dependent on the density of R5 peptides on the surface of the VNP; particles displaying 40 R5 peptides per CPMV (denoted as CPMV–R5H) interact strongly with the plasma membrane and are taken up into the cells via an energy-dependent mechanism while particles displaying 10 R5 peptides per CPMV (CPMV–R5L) are only slowly taken up. The fate of CPMV–R5 versus native CPMV particles within cells was evaluated in a co-localization time course study. It was indicated that the intracellular localization of CPMV–R5 and CPMV differs; CPMV remains trapped in Lamp-1 positive endolysosomes over long time frames; in contrast, 30–50% of the CPMV–R5 particles transitioned from the endosome into other cellular vesicles or compartments. Our data provide the groundwork for the development of efficient drug delivery formulations based on CPMV–R5.
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影响因子：
16.6
作者：
Dirksen, Anouk;Yegneswaran, Subramanian;Dawson, Philip E.
通讯作者：
Dawson, Philip E.
影响因子：
5.1
作者：
Khalil, I. A.;Kogure, K.;Harashima, H.
通讯作者：
Harashima, H.
影响因子：
2.5
作者：
Delehanty, James B.;Bradburne, Christopher E.;Medintz, Igor L.
通讯作者：
Medintz, Igor L.
影响因子：
10.8
作者：
Hooker, Jacob M.;Datta, Ankona;Francis, Matthew B.
通讯作者：
Francis, Matthew B.
影响因子：
3.4
作者：
Hällbrink, M;Florén, A;Langel, Ü
通讯作者：
Langel, Ü