Small-Interfering RNA-Eluting Surfaces as a Novel Concept for Intravascular Local Gene Silencing
Small-Interfering RNA-Eluting Surfaces as a Novel Concept for Intravascular Local Gene Silencing
复制标题
DOI：
10.2119/molmed.2011.00143
复制
发表时间：
2011-11-01
影响因子：
5.7
通讯作者：
Wendel, Hans Peter
中科院分区：
文献类型：
--
作者：
Nolte, Andrea;Walker, Tobias;Wendel, Hans Peter
新的药物洗脱支架（DES）方法最近已被证明可以改善血管内介入治疗的结局。一项新技术是设计基因沉默支架，该支架抑制特异性小干扰RNA（siRNA），以促进血管壁再生。尽管用于改变基因表达的siRNA在体外实验中已经超过了预期，但是siRNA的功能性和局部递送仍然是RNA干扰在体内应用的主要障碍。在这个初步的体外研究中，我们研究了一种表面固定的siRNA递送技术，该技术很容易适用于体内局部血管内应用。通过流式细胞术和定量实时聚合酶链反应在原代人内皮细胞中检查由与聚乙烯亚胺（PEI）复合的特异性siRNA组成的明胶涂层的转染效力。研究了几种培养基条件，如培养期间存在或不存在血清。此外，测试了不同siRNA和PEI量以及氮/磷酸盐比率的转染效率。由PEI和针对示例性内皮粘附分子受体的siRNA组成的明胶涂层实现了约70%的显著敲减。涂层的转染效率不受血清存在的影响。这项初步研究的结果支持了这样的预期，即这种新型涂层可能有利于局部体内基因沉默（例如，当固定在支架或球囊上进行经皮腔内冠状动脉成形术时）。然而，需要进一步的动物实验来证实转化为临床实践。这项有趣的技术为后DES时代的更复杂和个性化的涂层开辟了道路，使内膜增生途径中涉及的基因沉默。(C)2011 Feinstein医学研究所，www.feinsteininstitute.org在线地址：http://www.molmed.org doi：10.2119/molmed.2011.00143
New drug-eluting stent (DES) methods have recently been demonstrated to improve outcomes of intravascular interventions. A novel technique is the design of gene-silencing stents that elute specific small-interfering RNAs (siRNAs) for better vascular wall regeneration. Although siRNAs used to alter gene expression have surpassed expectations in in vitro experiments, the functional and local delivery of siRNAs is still the major obstacle for the in vivo application of RNA interference. In this preliminary in vitro study we investigated a surface-immobilized siRNA delivery technique that would be readily adaptable for local intravascular applications in vivo. The transfection potency of gelatin coatings consisting of a specific siRNA complexed with polyethylenimine (PEI) was examined in primary human endothelial cells by flow cytometry and quantitative real-time polymerase chain reaction. Several media conditions, such as the presence or absence of serum during cultivation, were investigated. Furthermore, different siRNA and PEI amounts, as well as nitrogen/phosphate ratios, were tested for their transfection efficiency. Gelatin coatings consisting of PEI and siRNA against an exemplary endothelial adhesion molecule receptor achieved a significant knockdown of around 70%. The transfection efficiency of the coatings was not influenced by the presence of serum. The results of this preliminary study support the expectation that this novel coating may be favorable for local in vivo gene silencing (for example, when immobilized on stents or balloons for percutanous transluminal coronary angioplasty). However, further animal experiments are needed to confirm the translation into clinical practice. This intriguing technology leads the way to more sophisticated and individualized coatings for the post-DES era, toward silencing of genes involved in the pathway of intimal hyperplasia. (C) 2011 The Feinstein Institute for Medical Research, www.feinsteininstitute.org Online address: http://www.molmed.org doi: 10.2119/molmed.2011.00143