Particle Tracking of Intracellular Trafficking of Octaarginine-modified Liposomes: A Comparative Study With Adenovirus
Particle Tracking of Intracellular Trafficking of Octaarginine-modified Liposomes: A Comparative Study With Adenovirus
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DOI：
10.1038/mt.2010.33
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发表时间：
2010-05-01
影响因子：
12.4
通讯作者：
Harashima, Hideyoshi
中科院分区：
文献类型：
--
作者：
Akita, Hidetaka;Enoto, Kaoru;Harashima, Hideyoshi
先前报道，八精氨酸（R8）修饰的脂质体（R8-Lip）通过巨胞饮作用被摄取，随后递送至核外围。在本研究中，我们研究了 R8-Lips 的细胞质转运机制，并与腺病毒进行比较。用微管破坏试剂（诺考达唑）处理比腺病毒更广泛地抑制封装 R8-Lip 的质粒 DNA (pDNA) 的转染活性。观察到 R8-Lips 沿着绿色荧光蛋白（GFP）标记的微管定向运输；然而，速度比不含 R8-Lips 的腺病毒或核内体慢。通过诺考达唑治疗，R8-Lips 中的这些定向运动被消除，而腺病毒继续进行随机运动。这一发现表明，R8-Lip 的核进入主要涉及微管依赖性运输，而明显的扩散运动也在腺病毒的核进入中发挥作用。此外，量子点标记的 pDNA 与罗丹明标记的脂质包膜同时进行定向运动，表明 R8-Lips 以完整的形式进行微管依赖性运输。载体和内体的双粒子追踪表明，R8-Lip 是定向运输的，与内体相关，而这发生在腺病毒中内体逃逸之后。总的来说，本文报道的结果表明囊泡运输是 R8-Lips 细胞质运输的关键因素。
It is previously reported that octaarginine (R8)-modified liposome (R8-Lip) was taken up via macropinocytosis, and subsequently delivered to the nuclear periphery. In the present study, we investigated the mechanism for the cytoplasmic transport of R8-Lips, comparing with that for adenovirus. Treatment with microtubule-disruption reagent (nocodazole) inhibited the transfection activity of plasmid DNA (pDNA)-encapsulating R8-Lip more extensively than that of adenovirus. The directional transport of R8-Lips along green fluorescent protein (GFP)-tagged microtubules was observed; however, the velocity was slower than those for adenovirus or endosomes that were devoid of R8-Lips. These directional motions were abrogated in R8-Lips by nocodazole treatment, whereas adenovirus continued to undergo random motion. This finding suggests that the nuclear access of R8-Lip predominantly involves microtubule-dependent transport, whereas an apparent diffusive motion is also operative in nuclear access of adenovirus. Furthermore, quantum dot-labeled pDNA underwent directional motion concomitantly with rhodamine-labeled lipid envelopes, indicating that the R8-Lips were subject to microtubule-dependent transport in the intact form. Dual particle tracking of carriers and endosomes revealed that R8-Lip was directionally transported, associated with endosomes, whereas this occurs after endosomal escape in adenovirus. Collectively, the findings reported herein indicate that vesicular transport is a key factor in the cytoplasmic transport of R8-Lips.