A novel peptide ligand-coated nano-siRNA-lipoplex technology for kidney targeted gene therapy.
A novel peptide ligand-coated nano-siRNA-lipoplex technology for kidney targeted gene therapy.
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发表时间：
2022
影响因子：
2.2
通讯作者：
Xia Liu;Jian-dong Zhang;Anna Tang;Liang Xu;Yufeng Huang
中科院分区：
文献类型：
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作者：
Xia Liu;Jian-dong Zhang;Anna Tang;Liang Xu;Yufeng Huang
小干扰RNA （siRNA）沉默特定疾病相关基因，有望用于肾脏疾病的治疗。然而，运送到预定的行动地点仍然是一个主要障碍。本研究的目的是开发肾小球特异性siRNA颗粒，用于肾脏疾病的靶向基因治疗。方法采用一种由siRNA组成的新型纳米颗粒系统，该系统由非抑制性纤溶酶原激活剂抑制剂1R （PAI-1R）包裹的阳离子脂质体（Lip）组成，选择性靶向肾小球细胞，并在肾病大鼠模型中使用转化生长因子-β1 (TGF-β1)-siRNA进行测试。结果在优化后的配比下，PAI-1R、Lip和siRNA形成了接近中性表面电荷（+5.63±1.45 mV）、尺寸相对均匀（68.9±4.73 nm）的致密纳米结构颗粒。当使用荧光标记的siRNA时，标记的siRNA纳米颗粒特异性地出现在肾小球中。靶向递送tgf - β1基因特异性siRNA可降低肾小球中升高的tgf - β1 mRNA表达和蛋白产生，但对肺、脾脏、动脉或肾髓质以及注射OX-7诱导的肾病大鼠的tgf - β1 mRNA水平无影响，持续时间长达5天。PAI-1R-Lip-TGF-β1 siRNA处理显著降低肾小球基质积累和PAI-1和纤维连接蛋白表达的增加。结论：单剂量PAI-1R-Lip-TGF-β1 siRNA抑制肾小球TGF-β1基因表达，从而特异性和有效地改善肾病大鼠肾小球硬化，而不影响大多数其他器官。靶基因沉默对肾小球疾病至关重要，这可能是一种有希望的肾脏疾病治疗策略。
OBJECTIVES Small interfering RNA (siRNA) that silences specific disease-related genes holds the promise for the treatment of renal disease. However, delivery to the intended site of action remains a major obstacle. The goal of this study was to develop glomerulus-specific siRNA particles for targeted gene therapy of kidney diseases. METHODS We used a novel nanoparticle-based system comprised of siRNA in cationic liposomes (Lip) coated with non-inhibitory plasminogen activator inhibitor 1R (PAI-1R) that selectively targets glomerular cells and tested it with transforming growth factor-beta 1 (TGF-β1)-siRNA in nephritic rat model. RESULTS At the optimized ratio of components, three of PAI-1R, Lip and siRNA formed the compact nanostructured particles with close to neutral surface charge (+5.63 ± 1.45 mV) and relatively uniform size (68.9 ± 4.73 nm). When the fluorescence-conjugated siRNA was used, the labeled siRNA nanoparticles appeared specifically in glomeruli. Targeted delivery of siRNA specific to the TGFβ1 gene reduced elevated TGFβ1 mRNA expression and protein production in glomeruli, but had no effect on TGFβ1 mRNA levels in lung, spleen, artery or renal medulla, and in nephritic rats induced by injection of OX-7, for up to 5 days. PAI-1R-Lip-TGF-β1 siRNA administration significantly reduced increases in glomerular matrix accumulation and expression of PAI-1 and fibronectin. CONCLUSIONS We conclude that a single dose of PAI-1R-Lip-TGF-β1 siRNA inhibited glomerular TGF-β1 gene expression thereby ameliorating glomerulosclerosis specifically and efficiently in nephritic rats without affecting most of other organs. The target silencing of genes critical for glomerular diseases may represent a promising treatment strategy for kidney disease.