College of Pharmacy, Yanbian University, Yanji 133000, Jilin Province, P.R. China.
Department of Radiology, Affiliated Hospital of Yanbian University, Yanji 133000, Jilin Province, P.R. China.
Curr Pharm Des. 2024;30(19):1507-1518. doi: 10.2174/0113816128258852231204102044.
Gene therapy has been widely concerned because of its unique therapeutic mechanism. However, due to the lack of safe and effective carries, it has not been widely used in clinical practice. Glypican 3 (GPC3) is a highly specific proteoglycan for hepatocellular carcinoma and is a potential diagnostic and therapeutic target for hepatocellular carcinoma. Herein, to monitor the effect of gene therapy and enhance the transfection efficiency of gene carriers, GPC3-modified lipid polyethyleneimine-modified superparamagnetic nanoparticle (GLPS), a type of visualized carrier for siRNA (small-interfering RNA) targeting the liver, was prepared.
We performed in vitro gene silencing, cytotoxicity, and agarose gel electrophoresis to identify the optimal GLPS formulation. MRI and Prussian blue staining verified the liver-targeting function of GLPS. We also analyzed the biocompatibility of GLPS by co-culturing with rabbit red blood cells. Morphological changes were evaluated using HE staining.
The GLPS optimal formulation consisted of LPS and siRNA at a mass ratio of 25:1 and LPS and DSPE-PEG-GPC3 at a molar ratio of 2:3. GLPS exhibited evident liver-targeting function. , we did not observe morphological changes in red blood cells or hemolysis after co-culture. In vivo, routine blood analysis revealed no abnormalities after GLPS injection. Moreover, the tissue morphology of the kidney, spleen, and liver was normal without injury or inflammation.
GLPS could potentially serve as an effective carrier for liver-targeted MRI monitoring and siRNA delivery.
基因治疗因其独特的治疗机制而受到广泛关注。然而，由于缺乏安全有效的载体，尚未广泛应用于临床实践。磷脂酰聚糖 3（GPC3）是肝细胞癌的高度特异性蛋白聚糖，是肝细胞癌潜在的诊断和治疗靶点。在此，为了监测基因治疗的效果并提高基因载体的转染效率，制备了 GPC3 修饰的脂质聚乙烯亚胺修饰的超顺磁性纳米颗粒（GLPS），这是一种针对肝脏的靶向 siRNA（小干扰 RNA）的可视化载体。
我们进行了体外基因沉默、细胞毒性和琼脂糖凝胶电泳实验，以确定最佳的 GLPS 配方。MRI 和普鲁士蓝染色验证了 GLPS 的肝靶向功能。我们还通过与兔红细胞共培养来分析 GLPS 的生物相容性。通过 HE 染色评估形态变化。
GLPS 的最佳配方由 LPS 和 siRNA 以质量比 25:1 和 LPS 和 DSPE-PEG-GPC3 以摩尔比 2:3 组成。GLPS 表现出明显的肝靶向功能。我们没有观察到共培养后红细胞的形态变化或溶血。在体内，GLPS 注射后常规血液分析无异常。此外，肾脏、脾脏和肝脏的组织形态正常，无损伤或炎症。
GLPS 可能成为用于肝靶向 MRI 监测和 siRNA 递送的有效载体。
