Intranasal drug delivery of small interfering RNA targeting Beclin1 encapsulated with polyethylenimine (PEI) in mouse brain to achieve HIV attenuation.
Intranasal drug delivery of small interfering RNA targeting Beclin1 encapsulated with polyethylenimine (PEI) in mouse brain to achieve HIV attenuation.
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DOI：
10.1038/s41598-017-01819-9
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发表时间：
2017-05-12
影响因子：
4.6
通讯作者：
El-Hage N
中科院分区：
文献类型：
--
作者：
Rodriguez M;Lapierre J;Ojha CR;Kaushik A;Batrakova E;Kashanchi F;Dever SM;Nair M;El-Hage N
我们以前报道，激活宿主自噬蛋白，Beclin 1，由HIV-1感染代表了一个重要的机制，在控制艾滋病毒复制和病毒诱导的炎症反应的小胶质细胞。现有的抗逆转录病毒治疗方法在有效穿过血脑屏障以及识别和选择性消除持续感染艾滋病毒的脑储库方面的能力有限。在本研究中，第一次，在成年小鼠脑中的生物分布和疗效的非侵入性鼻内递送的小干扰RNA（siRNA）对Beclin 1基因使用阳离子线性聚乙烯亚胺（PEI）作为基因载体进行了研究。在递送后4和24小时，在前额叶皮质的神经元和神经胶质细胞的细胞质中发现鼻内递送的异硫氰酸荧光素（FITC）标记的对照siRNA，没有遇到重大的不良免疫反应。鼻内递送靶向Beclin 1的siRNA显著地耗尽了脑组织中的靶蛋白表达水平，没有毒性证据。通过拉曼光谱表征并证实siRNA与PEI-聚合物的结合。这些结果表明，鼻内药物递送允许将PEI-siRNA纳米复合物直接递送至中枢神经系统，这可能为HIV感染的脑中的基因沉默介导的治疗提供有效手段。
We previously reported that activation of the host autophagic protein, Beclin1, by HIV-1 infection represents an essential mechanism in controlling HIV replication and viral-induced inflammatory responses in microglial cells. Existing antiretroviral therapeutic approaches have been limited in their ability to cross the blood-brain barrier effectively and recognize and selectively eliminate persistent HIV-infected brain reservoirs. In the present study and for the first time, the bio-distribution and efficacy of noninvasive intranasal delivery of small interfering RNA (siRNA) against the Beclin1 gene using the cationic linear polyethylenimines (PEI) as a gene carrier was investigated in adult mouse brain. Fluorescein isothiocyanate (FITC)-labeled control siRNA delivered intranasally was found in the cytoplasm of neurons and glial cells of the prefrontal cortex at 4 and 24 hours post-delivery, with no major adverse immune reaction encountered. Intranasal delivery of the siRNA targeting Beclin1 significantly depleted the target protein expression levels in brain tissues with no evidence of toxicity. Binding of siRNA to PEI-polymer was characterized and confirmed by Raman spectroscopy. These results indicate that the intranasal drug delivery allows for the direct delivery of the PEI-siRNA nano-complex to the central nervous system, which could potentially offer an efficient means of gene silencing-mediated therapy in the HIV-infected brain.