Yin Xing, Harmancey Romain, Frierson Brion, Wu Jean G, Moody Melanie R, McPherson David D, Huang Shao-Ling
Division of Cardiology, Department of Internal Medicine, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA.
Department of Diagnostic Sciences, The University of Texas Health Science Center at Houston, Houston, TX 77054, USA.
Pharmaceutics. 2024 Feb 29;16(3):343. doi: 10.3390/pharmaceutics16030343.
Liposomes as carriers for CRISPR/Cas9 complexes represent an attractive approach for cardiovascular gene therapy. A critical barrier to this approach remains the efficient delivery of CRISPR-based genetic materials into cardiomyocytes. Echogenic liposomes (ELIP) containing a fluorescein isothiocyanate-labeled decoy oligodeoxynucleotide against nuclear factor kappa B (ELIP-NF-κB-FITC) were used both in vitro on mouse neonatal ventricular myocytes and in vivo on rat hearts to assess gene delivery efficacy with or without ultrasound. In vitro analysis was then repeated with ELIP containing Cas9-sg-IL1RL1 (interleukin 1 receptor-like 1) RNA to determine the efficiency of gene knockdown. ELIP-NF-κB-FITC without ultrasound showed limited gene delivery in vitro and in vivo, but ultrasound combined with ELIP notably improved penetration into heart cells and tissues. When ELIP was used to deliver Cas9-sg-IL1RL1 RNA, gene editing was successful and enhanced by ultrasound. This innovative approach shows promise for heart disease gene therapy using CRISPR technology.
脂质体作为CRISPR/Cas9复合物的载体，是心血管疾病基因治疗的一种有吸引力的方法。这种方法的一个关键障碍仍然是如何将基于CRISPR的遗传物质有效地递送至心肌细胞。含有针对核因子κB的异硫氰酸荧光素标记的诱饵寡脱氧核苷酸的可回声脂质体（ELIP），在体外用于小鼠新生心室肌细胞，在体内用于大鼠心脏，以评估有无超声情况下的基因递送效果。然后，使用含有Cas9-sg-IL1RL1（白细胞介素1受体样1）RNA的ELIP重复体外分析，以确定基因敲低效率。无超声的ELIP-NF-κB-FITC在体外和体内均显示出有限的基因递送，但超声与ELIP结合显著提高了对心脏细胞和组织的穿透能力。当使用ELIP递送Cas9-sg-IL1RL1 RNA时，基因编辑成功，且超声增强了编辑效果。这种创新方法显示出利用CRISPR技术进行心脏病基因治疗的潜力。
