Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, USA.
Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, USA; Dept. of Pediatrics, University of Washington, Seattle, USA.
J Control Release. 2014 May 28;182:111-20. doi: 10.1016/j.jconrel.2014.03.002. Epub 2014 Mar 18.
Ultrasound (US)-mediated gene delivery has emerged as a promising non-viral method for safe and selective gene delivery. When enhanced by the cavitation of microbubbles (MBs), US exposure can induce sonoporation that transiently increases cell membrane permeability for localized delivery of DNA. The present study explores the effect of generalizable MB customizations on MB facilitation of gene transfer compared to Definity®, a clinically available contrast agent. These modifications are 1) increased MB shell acyl chain length (RN18) for elevated stability and 2) addition of positive charge on MB (RC5K) for greater DNA associability. The MB types were compared in their ability to facilitate transfection of luciferase and GFP reporter plasmid DNA in vitro and in vivo under various conditions of US intensity, MB dosage, and pretreatment MB-DNA incubation. The results indicated that both RN18 and RC5K were more efficient than Definity®, and that the cationic RC5K can induce even greater transgene expression by increasing payload capacity with prior DNA incubation without compromising cell viability. These findings could be applied to enhance MB functions in a wide range of therapeutic US/MB gene and drug delivery approach. With further designs, MB customizations have the potential to advance this technology closer to clinical application.
超声（US）介导的基因传递已成为一种有前途的非病毒方法，用于安全和选择性的基因传递。当微泡（MB）的空化增强时，US 暴露可以诱导声孔，瞬时增加细胞膜通透性，实现 DNA 的局部传递。本研究探讨了可推广的 MB 定制对基因转染的 MB 促进作用与 Definity®（一种临床可用的造影剂）的比较。这些修饰是 1）增加 MB 壳酰链长度（RN18）以提高稳定性，2）在 MB 上添加正电荷（RC5K）以提高 DNA 亲和力。比较了这些 MB 类型在不同 US 强度、MB 剂量和预处理 MB-DNA 孵育条件下，体外和体内促进荧光素酶和 GFP 报告质粒 DNA 转染的能力。结果表明，RN18 和 RC5K 都比 Definity®更有效，而阳离子 RC5K 通过在 DNA 孵育前增加有效载荷能力，即使在不损害细胞活力的情况下，也可以诱导更大的转基因表达。这些发现可应用于增强广泛的治疗性 US/MB 基因和药物输送方法中的 MB 功能。通过进一步的设计，MB 定制有可能使这项技术更接近临床应用。
