Laboratory of Biomaterials and Translational Medicine, Center for Nanomedicine, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510630, China.
Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO, 63110, USA.
Biomaterials. 2023 Nov;302:122349. doi: 10.1016/j.biomaterials.2023.122349. Epub 2023 Oct 6.
Targeting the activated epidermal growth factor receptor (EGFR) via clustered regularly interspaced short palindromic repeat (CRISPR) technology is appealing to overcome the drug resistance of hepatocellular carcinoma (HCC) towards tyrosine kinase inhibitor (TKI) therapy. However, combining these two distinct drugs using traditional liposomes results in a suboptimal synergistic anti-HCC effect due to the limited CRISPR/Cas9 delivery efficiency caused by lysosomal entrapment after endocytosis. Herein, we developed a liver-targeting gene-hybridizing-TKI fusogenic liposome (LIGHTFUL) that can achieve high CRISPR/Cas9 expression to reverse the EGFR-mediated drug resistance for enhanced TKI-based HCC therapy efficiently. Coated with a galactose-modified membrane-fusogenic lipid layer, LIGHTFUL reached the targeting liver site to fuse with HCC tumor cells, directly and efficiently transporting interior CDK5- and PLK1-targeting CRISPR/Cas9 plasmids (pXG333-CPs) into the HCC cell cytoplasm and then the cell nucleus for efficient expression. Such membrane-fusion-mediated pXG333-CP delivery resulted in effective downregulation of both CDK5 and PLK1, sufficiently inactivating EGFR to improve the anti-HCC effects of the co-delivered TKI, lenvatinib. This membrane-fusion-participant codelivery strategy optimized the synergetic effect of CRISPR/Cas9 and TKI combinational therapy as indicated by the 0.35 combination index in vitro and the dramatic reduction of subcutaneous and orthotopic TKI-insensitive HCC tumor growth in mice. Therefore, the established LIGHTFUL provides a unique co-delivery platform to combine gene editing and TKI therapies for enhanced synergetic therapy.
通过成簇规律间隔短回文重复 (CRISPR) 技术靶向激活的表皮生长因子受体 (EGFR)，以克服肝细胞癌 (HCC) 对酪氨酸激酶抑制剂 (TKI) 治疗的耐药性，这是一种很有吸引力的方法。然而，由于内吞后溶酶体捕获导致 CRISPR/Cas9 递送效率有限，使用传统脂质体将这两种截然不同的药物结合在一起，导致联合抗 HCC 效果不佳。在此，我们开发了一种肝靶向基因杂交-TKI 融合脂质体 (LIGHTFUL)，它可以实现高效的 CRISPR/Cas9 表达，逆转 EGFR 介导的耐药性，从而有效增强基于 TKI 的 HCC 治疗效果。LIGHTFUL 被一层经过半乳糖修饰的膜融合脂质层所包裹，到达靶向肝脏部位与 HCC 肿瘤细胞融合，直接高效地将内部靶向 CDK5 和 PLK1 的 CRISPR/Cas9 质粒 (pXG333-CPs) 递送至 HCC 细胞质，然后递送至细胞核进行高效表达。这种膜融合介导的 pXG333-CP 递呈导致 CDK5 和 PLK1 的有效下调，充分使 EGFR 失活，从而提高共递呈的 TKI（仑伐替尼）的抗 HCC 效果。这种膜融合参与者共递呈策略优化了 CRISPR/Cas9 和 TKI 联合治疗的协同效应，体外 0.35 的联合指数和对小鼠皮下和原位 TKI 不敏感 HCC 肿瘤生长的显著减少均证明了这一点。因此，所建立的 LIGHTFUL 提供了一个独特的共递呈平台，将基因编辑和 TKI 治疗相结合，以增强协同治疗效果。
