Ren Xiao-He, He Xiao-Yan, Liu Bo-Ya, Xu Chang, Cheng Si-Xue
Key Laboratory of Biomedical Polymers of Ministry of Education, Department of Chemistry, Wuhan University, Wuhan 430072, People's Republic of China.
ACS Appl Bio Mater. 2020 Nov 16;3(11):7831-7839. doi: 10.1021/acsabm.0c01009. Epub 2020 Nov 5.
Gene delivery vectors possess critical roles in effective genome editing. In this study, a multiple functional vector for encapsulating CRISPR/Cas9 plasmid was designed to knock out PPM1D gene and prevent cancer malignancy. The plasmid was complexed with a KALA peptide with the capability of endosomal escape and histones for nuclear transportation and then decorated by hyaluronic acid (HA) and AS1411-incorporated hyaluronic acid (AHA) targeting CD44 and nucleolin overexpressed in cancer cells to form AHA/HA/KALA/histone/plasmid nanoparticles. The constructed multifunctional plasmid delivery system with the cancer targeting specificity can realize efficient genome editing for PPM1D knockout and thus dramatically downregulate PPM1D expression in targeted malignant cells. More importantly, PPM1D knockout results in upregulation of p21 and p-p38 as well as downregulation of cyclin D1, MMP9, CYR61, and vimentin. The edited cancer cells exhibit suppressed proliferation, migration, and invasion, indicating the successful reversal of tumor malignancy.
基因递送载体在有效的基因组编辑中起着关键作用。在本研究中，设计了一种用于封装CRISPR/Cas9质粒的多功能载体，以敲除PPM1D基因并预防癌症恶性进展。该质粒与具有内体逃逸能力的KALA肽和用于核转运的组蛋白复合，然后用透明质酸（HA）和掺入AS1411的透明质酸（AHA）进行修饰，AHA靶向癌细胞中过表达的CD44和核仁素，以形成AHA/HA/KALA/组蛋白/质粒纳米颗粒。构建的具有癌症靶向特异性的多功能质粒递送系统可实现对PPM1D敲除的高效基因组编辑，从而显著下调靶向恶性细胞中PPM1D的表达。更重要的是，PPM1D敲除导致p21和p-p38上调以及细胞周期蛋白D1、基质金属蛋白酶9、细胞外调节蛋白激酶61和波形蛋白下调。编辑后的癌细胞表现出增殖、迁移和侵袭受到抑制，表明肿瘤恶性进展成功逆转。
