Application of Deacetylated Poly-N-Acetyl Glucosamine Nanoparticles for the Delivery of miR-126 for the Treatment of Cecal Ligation and Puncture-Induced Sepsis
Application of Deacetylated Poly-N-Acetyl Glucosamine Nanoparticles for the Delivery of miR-126 for the Treatment of Cecal Ligation and Puncture-Induced Sepsis
复制标题
DOI：
10.1007/s10753-018-0882-8
复制
发表时间：
2019-02-01
期刊：
影响因子：
5.1
通讯作者：
Fan, Hongkuan
中科院分区：
文献类型：
--
作者：
Buie, Joy N. Jones;Zhou, Yue;Fan, Hongkuan
脓毒症是一种急性炎症综合征的感染。在某些情况下，败血症引起的过度炎症导致内皮功能障碍和随后的血管通透性增加，导致器官衰竭。我们先前表明，用高表达microRNA-126（miR-126）的内皮祖细胞治疗可改善盲肠结扎穿孔（CLP）脓毒症小鼠的存活率。miRNA是基因表达和细胞功能的重要调节因子，在内皮稳态中起主要作用，并且可能代表新兴的治疗方式。然而，由于普遍存在的RNA酶的快速降解，在体外和体内将miRNA递送至细胞是具有挑战性的。在此，我们开发了一种纳米颗粒递送系统，分别将脱乙酰化聚-N-乙酰基葡糖胺（DEAC-pGlcNAc）聚合物与miRNA-126- 3 p和miRNA-126- 5 p组合，并在体外和体内测试这些组合。我们的结果表明，DEAC-pGlcNAc聚合物具有合适的大小和zeta电位，用于细胞摄取，并且当复合时，DEAC-pGlcNAc保护miRNA免受RNA酶A降解。此外，DEAC-pGlcNAc有效地包封miRNA，如通过防止它们在琼脂糖凝胶中迁移所证明的。DEAC-pGlcNAc-miRNA复合物被多种细胞类型摄取，并且递送的miRNA在体外对其靶标（包括pERK和DLK-1）具有生物学效应。此外，我们发现，递送单独的DEAC-pGlcNAc或DEAC-pGlcNAc：miRNA-126- 5 p纳米颗粒至脓毒症动物显著提高了存活率，保持了血管完整性，并调节了细胞因子的产生。这些复合研究支持DEAC-pGlcNAc纳米颗粒是递送miRNA的有效平台并且它们可以在脓毒症中提供治疗益处的概念。
Sepsis is an acute inflammatory syndrome in response to infection. In some cases, excessive inflammation from sepsis results in endothelial dysfunction and subsequent increased vascular permeability leading to organ failure. We previously showed that treatment with endothelial progenitor cells, which highly express microRNA-126 (miR-126), improved survival in mice subjected to cecal ligation and puncture (CLP) sepsis. miRNAs are important regulators of gene expression and cell function, play a major role in endothelial homeostasis, and may represent an emerging therapeutic modality. However, delivery of miRNAs to cells in vitro and in vivo is challenging due to rapid degradation by ubiquitous RNases. Herein, we developed a nanoparticle delivery system separately combining deacetylated poly-N-acetyl glucosamine (DEAC-pGlcNAc) polymers with miRNA-126-3p and miRNA-126-5p and testing these combinations in vitro and in vivo. Our results demonstrate that DEAC-pGlcNAc polymers have an appropriate size and zeta potential for cellular uptake and when complexed, DEAC-pGlcNAc protects miRNA from RNase A degradation. Further, DEAC-pGlcNAc efficiently encapsulates miRNAs as evidenced by preventing their migration in an agarose gel. The DEAC-pGlcNAc-miRNA complexes were taken up by multiple cell types and the delivered miRNAs had biological effects on their targets in vitro including pERK and DLK-1. In addition, we found that delivery of DEAC-pGlcNAc alone or DEAC-pGlcNAc:miRNA-126-5p nanoparticles to septic animals significantly improved survival, preserved vascular integrity, and modulated cytokine production. These composite studies support the concept that DEAC-pGlcNAc nanoparticles are an effective platform for delivering miRNAs and that they may provide therapeutic benefit in sepsis.