通过脂质纳米颗粒递送的 PLK1 靶向 siRNA 在 HBV 感染的肝细胞中的抗病毒活性

IF 1.3 4区 医学 Q4 INFECTIOUS DISEASES
Antiviral Therapy Pub Date : 2020-01-01 DOI:10.3851/IMP3361
Adrien Foca, Ammen Dhillon, Thomas Lahlali, Julie Lucifora, Anna Salvetti, Michel Rivoire, Amy Lee, David Durantel
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引用次数: 7

摘要

背景:在 HBV 驱动的癌变过程中,HBV 与 PLK1 之间的联系已得到明确证实,而且我们最近还发现 PLK1 是 HBV 感染早期阶段的病毒因子。此外,我们已经证明,小分子 PLK1 抑制剂 BI-2536 能非常有效地抑制 HBV DNA 的新合成,特别是通过调节 HBc 磷酸化来影响核壳组装。然而,由于小分子激酶抑制剂通常选择性较差,因此需要一种更特异、更安全的靶向 PLK1 的策略,以开发抗慢性 HBV 感染的潜在药物。我们使用标准检测方法来监测 LNP siPLK1 或对照组(LNP siHBV 和 LNP siNon-targeting)对 HBV 复制和细胞活力的影响:低至 100 ng/ml 剂量的 LNP-siPLK1 可使分泌的 HBV DNA(病毒颗粒)减少 75%以上,与 LNP siHBV 或 10 µM 替诺福韦(TFV)的效果相当,但不影响细胞活力。有趣的是,与使用 TFV 时的结果相反,在 LNP siPLK1 处理中也观察到了对病毒 RNA 和 HBe/HBsAg 分泌的强烈抑制。这与细胞内 vRNA 积累的显著减少有关,而这种减少与cccDNA 水平的任何变化无关,因此表明这是一种转录或转录后调节作用。通过生化方法抑制 PLK1 并未获得这种效果,这表明 PLK1 的作用与酶无关:本研究强调,特异性 PLK1 抑制有助于改善慢性阻塞性肺病患者的 HBsAg 消失情况,很可能与其他 HBsAg 靶向策略相结合。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Antiviral activity of PLK1-targeting siRNA delivered by lipid nanoparticles in HBV-infected hepatocytes.

Background: A link between HBV and PLK1 was clearly evidenced in HBV-driven carcinogenesis, and we have also recently shown that PLK1 is a proviral factor in the early phases of HBV infection. Moreover, we have shown that BI-2536, a small molecule PLK1 inhibitor, was very efficient at inhibiting HBV DNA neosynthesis, notably by affecting nucleocapsid assembly as a result of the modulation of HBc phosphorylation. Yet, as small molecule kinase inhibitors often feature poor selectivity, a more specific and safer strategy to target PLK1 would be needed for a potential development against chronic HBV infections.

Methods: Here, we analysed using both freshly isolated primary human hepatocytes and differentiated HepaRG, the anti-HBV properties of an LNP-encapsulated PLK1-targeting siRNA. Standard assays were used to monitor the effect of LNP siPLK1, or controls (LNP siHBV and LNP siNon-targeting), on HBV replication and cell viability.

Results: A dose as low as 100 ng/ml of LNP-siPLK1 resulted in a >75% decrease in secreted HBV DNA (viral particles), which was comparable to that obtained with LNP siHBV or 10 µM of tenofovir (TFV), without affecting cell viability. Interestingly, and in contrast to that obtained with TFV, a strong inhibition of viral RNA and HBe/HBsAg secretions was also observed under LNP siPLK1 treatment. This correlated with a significant intracellular decrease of vRNA accumulation, which was independent of any change in cccDNA levels, thus suggesting a transcriptional or post-transcriptional modulation. Such an effect was not obtained with a biochemical approach of PLK1 inhibition, suggesting an enzymatic-independent role of PLK1.

Conclusions: This study emphasizes that a specific PLK1 inhibition could help in achieving an improved HBsAg loss in CHB patients, likely in combination with other HBsAg-targeting strategies.

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来源期刊
Antiviral Therapy
Antiviral Therapy 医学-病毒学
CiteScore
2.60
自引率
8.30%
发文量
35
审稿时长
4-8 weeks
期刊介绍: Antiviral Therapy (an official publication of the International Society of Antiviral Research) is an international, peer-reviewed journal devoted to publishing articles on the clinical development and use of antiviral agents and vaccines, and the treatment of all viral diseases. Antiviral Therapy is one of the leading journals in virology and infectious diseases. The journal is comprehensive, and publishes articles concerning all clinical aspects of antiviral therapy. It features editorials, original research papers, specially commissioned review articles, letters and book reviews. The journal is aimed at physicians and specialists interested in clinical and basic research.
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