Tran Dominic M, Zhang Feng, Morrison Kyle P, Loeb Keith R, Harrang James, Kajimoto Masaki, Chavez Francisco, Wu Li, Miao Carol H
Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, WA 98101, USA.
Department of Radiology, University of Washington, Seattle, WA 98195, USA.
Mol Ther Methods Clin Dev. 2019 Jul 26;14:275-284. doi: 10.1016/j.omtm.2019.07.005. eCollection 2019 Sep 13.
Ultrasound (US)-mediated gene delivery (UMGD) of nonviral vectors was demonstrated in this study to be an effective method to transfer genes into the livers of large animals via a minimally invasive approach. We developed a transhepatic venous nonviral gene delivery protocol in combination with transcutaneous, therapeutic US (tUS) to facilitate significant gene transfer in pig livers. A balloon catheter was inserted into the pig hepatic veins of the target liver lobes via jugular vein access under fluoroscopic guidance. tUS exposure was continuously applied to the lobe with simultaneous infusion of pGL4 plasmid (encoding a luciferase reporter gene) and microbubbles. tUS was delivered via an unfocused, two-element disc transducer (H105) or a novel focused, single-element transducer (H114). We found applying transcutaneous US using H114 and H105 with longer pulses and reduced acoustic pressures resulted in an over 100-fold increase in luciferase activity relative to untreated lobes. We also showed effective UMGD by achieving focal regions of >10 relative light units (RLUs)/mg protein with minimal tissue damage, demonstrating the feasibility for clinical translation of this technique to treat patients with genetic diseases.
本研究证明，超声（US）介导的非病毒载体基因递送（UMGD）是一种通过微创方法将基因导入大型动物肝脏的有效方法。我们开发了一种经肝静脉非病毒基因递送方案，结合经皮治疗性超声（tUS），以促进猪肝脏中的显著基因转移。在荧光透视引导下，通过颈静脉通路将球囊导管插入目标肝叶的猪肝静脉中。在持续输注pGL4质粒（编码荧光素酶报告基因）和微泡的同时，对肝叶持续进行tUS照射。tUS通过非聚焦双元件圆盘换能器（H105）或新型聚焦单元件换能器（H114）进行递送。我们发现，使用H114和H105进行经皮超声，采用更长的脉冲并降低声压，相对于未处理的肝叶，荧光素酶活性增加了100倍以上。我们还通过在最小组织损伤的情况下实现>10相对光单位（RLUs）/mg蛋白质的聚焦区域，展示了有效的UMGD，证明了该技术临床转化用于治疗遗传疾病患者的可行性。
