On the mechanism of tissue-selective gene delivery by lipid nanoparticles
On the mechanism of tissue-selective gene delivery by lipid nanoparticles
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DOI：
10.1016/j.jconrel.2023.03.052
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发表时间：
2023-10-09
影响因子：
10.8
通讯作者：
Harashima，Hideyoshi
中科院分区：
文献类型：
--
作者：
Kimura，Seigo;Harashima，Hideyoshi
核酸纳米医学时代已经到来，Patisiran、一种小干扰 RNA (siRNA) 封装的脂质纳米颗粒 (LNP) 和用于 COVID-19 疫苗的 mRNA 负载 LNP 就证明了这一点。在 II/III 期临床试验中测试的用于递送核酸分子的纳米设计的多样性反映了这些技术的潜力。非病毒基因传递方面的这些突破，包括 LNP 的使用，引起了全世界对开发更有效药物的浓厚兴趣。该领域的下一步是针对肝脏以外的组织，这需要大量的研究工作和材料开发。然而，缺乏这方面的机制研究。本研究比较了两种类型的具有不同组织选择性的 LNP 来递送质粒 DNA (pDNA)，一种是肝脏选择性的，另一种是脾选择性的，旨在了解导致所递送基因的基因表达差异的机制。尽管基因表达存在 100-1000 倍的差异，但我们观察到这两种 LNP 的生物分布几乎没有差异。然后，我们通过定量实时 PCR (qPCR) 定量每个组织中递送的 pDNA 和 mRNA 表达量，以评估各种细胞内过程，例如核递送、转录和翻译。结果显示，翻译步骤存在 >100 倍的差异，但两次 LNP 递送到细胞核的 pDNA 量或 mRNA 表达量几乎没有差异。我们的研究结果表明，内源因素影响基因表达效率，而不是生物分布的程度。
The era of nucleic acid nanomedicine has arrived, as evidenced by Patisiran, a small interfering RNA (siRNA) encapsulated lipid nanoparticle (LNP), and mRNA-loaded LNPs used in COVID-19 vaccines. The diversity of nano-designs for delivering nucleic acid molecules tested in Phase II/III clinical trials reflects the potential of these technologies. These breakthroughs in non-viral gene delivery, including the use of LNPs, have attracted substantial interest worldwide for developing more effective drugs. A next step in this field is to target tissues other than the liver, which requires significant research efforts and material development. However, mechanistic studies in this area are lacking. This study compares two types of LNPs with different tissue-selectivity for delivering plasmid DNA (pDNA), one being liver-selective and the other spleen-selective, in an effort to understand the mechanisms responsible for differences in gene expression of delivered genes. We observed little difference in the biodistribution of these two LNPs despite the 100–1000-fold differences in gene expression. We then quantified the amount of delivered pDNA and mRNA expression in each tissue by quantitative real-time PCR (qPCR) to evaluate various intracellular processes, such as nuclear delivery, transcription and translation. The results showed a >100-fold difference in the translation step but there were little differences in amount of pDNA delivered to the nucleus or the amount of mRNA expression for the two LNP deliveries. Our findings suggest that endogenous factors affect gene expression efficiency not the extent of biodistribution.