Magnetically responsive biodegradable nanoparticles enhance adenoviral gene transfer in cultured smooth muscle and endothelial cells.
Magnetically responsive biodegradable nanoparticles enhance adenoviral gene transfer in cultured smooth muscle and endothelial cells.
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DOI：
10.1021/mp900017m
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发表时间：
2009-09
影响因子：
4.9
通讯作者：
Levy RJ
中科院分区：
文献类型：
--
作者：
Chorny M;Fishbein I;Alferiev I;Levy RJ
复制缺陷型腺病毒（Ad）载体已显示出作为基于基因递送的治疗应用的工具的前景。然而，它们的临床应用受到表达低水平柯萨奇-Ad受体（CAR）（负责病毒进入细胞的主要受体）的细胞类型中治疗转导水平不理想以及全身不良反应的限制。因此，Ad 与可生物降解的磁响应纳米颗粒 (MNP) 复合可实现靶向递送，可能有助于提高这些载体的安全性和效率。我们的假设是，磁驱动传递与可生物降解的 MNP 亲和力结合的 Ad 可以显着增加培养物中 CAR 缺陷血管细胞的转基因表达。荧光标记的 MNP 由含有氧化铁的聚丙交酯配制而成，并用 CAR 的 D1 结构域作为亲和连接体进行表面修饰。分别使用 540 nm/575 nm 和 485 nm/535 nm 的 λex/λem 对培养的内皮细胞和平滑肌细胞中的 MNP 细胞摄取和 GFP 报告基因转基因表达进行荧光分析。 Ad 与 MNP 的稳定载体特异性关联导致 MNP-Ad 复合物的形成，在短暂暴露于高梯度磁场后，显示出快速的细胞结合动力学，与自由载体或非磁性对照处理相比，所得基因转移水平显着增加。多元回归分析表明 MNP-Ad 介导的转导机制不同于游离 Ad，并证实了复合物对磁性条件下基因转移的主要贡献。在不影响细胞活力或生长动力学的情况下实现了磁增强转导。通过与可生物降解的 MNP 亲和络合来增强腺病毒基因递送是一种有前途的方法，有可能扩展病毒基因治疗策略的适用性。
Replication-defective adenoviral (Ad) vectors have shown promise as a tool for gene delivery-based therapeutic applications. Their clinical use is however limited by therapeutically suboptimal transduction levels in cell types expressing low levels of Coxsackie-Ad receptor (CAR), the primary receptor responsible for the cell entry of the virus, and by systemic adverse reactions. Targeted delivery achievable with Ad complexed with biodegradable magnetically responsive nanoparticles (MNP) may therefore be instrumental for improving both the safety and efficiency of these vectors. Our hypothesis was that magnetically driven delivery of Ad affinity-bound to biodegradable MNP can substantially increase transgene expression in CAR deficient vascular cells in culture. Fluorescently labeled MNP were formulated from polylactide with inclusion of iron oxide and surface-modified with the D1 domain of CAR as an affinity linker. MNP cellular uptake and GFP reporter transgene expression were assayed fluorimetrically in cultured endothelial and smooth muscle cells using λex/λem of 540 nm/575 nm and 485 nm/535 nm, respectively. Stable vector-specific association of Ad with MNP resulted in formation of MNP–Ad complexes displaying rapid cell binding kinetics following a brief exposure to a high gradient magnetic field with resultant gene transfer levels significantly increased compared to free vector or nonmagnetic control treatment. Multiple regression analysis suggested a mechanism of MNP–Ad mediated transduction distinct from that of free Ad, and confirmed the major contribution of the complexes to the gene transfer under magnetic conditions. The magnetically enhanced transduction was achieved without compromising the cell viability or growth kinetics. The enhancement of adenoviral gene delivery by affinity complexation with biodegradable MNP represents a promising approach with a potential to extend the applicability of the viral gene therapeutic strategies.
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影响因子：
12.4
作者：
Chorny, Michael;Fishbein, Ilia;Levy, Robert J.
通讯作者：
Levy, Robert J.
影响因子：
3.7
作者：
Mok, Hyejung;Park, Ji Won;Park, Tae Gwan
通讯作者：
Park, Tae Gwan
影响因子：
3.7
作者：
Pandori, MW;Hobson, DA;Sano, T
通讯作者：
Sano, T
DOI：
10.1046/j.1432-1327.2000.01606.x
发表时间：
2000-09-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
O'Brien, J;Wilson, I;Pognan, F
通讯作者：
Pognan, F
影响因子：
12.4
作者：
Mah, C;Fraites, TJ;Byrne, BJ
通讯作者：
Byrne, BJ