State Key Laboratory for Modification of Chemical Fibers and Polymer Materials, Shanghai Engineering Research Center of Nano-Biomaterials and Regenerative Medicine, College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620, People's Republic of China.
Department of Chemistry, College of Natural and Computational Sciences, University of Gondar, Gondar 196, Ethiopia.
ACS Appl Bio Mater. 2021 Sep 20;4(9):7168-7175. doi: 10.1021/acsabm.1c00743. Epub 2021 Aug 23.
Preparation of versatile and safe nanovectors for efficient cancer gene therapy remains to be challenging in the current nanomedicine. Herein, we report the formation of dendrimer nanoclusters for enhanced gene delivery toward gene therapy of cancer. Here, poly(amidoamine) (PAMAM) dendrimers of generation 3 (G3) were cross-linked with 4,4'-dithiodibutryic acid (DA) to form nanoclusters (NCs) through 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-induced covalent bonding. The synthesized G3-DA NCs having a hydrodynamic size of 219.3 nm possess good colloidal stability and can condense pDNA, encoding both enhanced green fluorescent protein and tumor suppressor p53 gene to form polyplexes with good cytocompatibility. Strikingly, the created NCs/pDNA polyplexes enable 2.3 and 2.1 times higher gene transfection to cancer cells than the counterpart materials of single G3 and G5 PAMAM dendrimers, respectively, under the same conditions. Furthermore, polyplex-treated cancer cells have upregulated p53 and p21 protein and mRNA expression levels and downregulated Cyclin-D1 and CDK-4 protein and mRNA expressions, thus arresting the cell cycle to the G1 phase to achieve cancer cell gene therapy. The gene delivery efficiency of the polyplexes was further validated through the tumor therapy without systemic toxicity. The synthesized highly efficient dendrimer NC-based vector system with low cytotoxicity may be extended to tackle various types of diseases related to genetic disorders.
制备用于高效癌症基因治疗的多功能和安全的纳米载体仍然是当前纳米医学中的挑战。在此，我们报告了树枝状大分子纳米簇的形成，用于增强癌症基因治疗的基因传递。在这里，通过 1-（3-二甲基氨基丙基）-3-乙基碳二亚胺盐酸盐诱导的共价键合，将第三代（G3）聚酰胺-胺（PAMAM）树枝状大分子与 4,4'-二硫代二丁酸（DA）交联形成纳米簇（NC）。合成的 G3-DA NC 具有 219.3nm 的水动力粒径，具有良好的胶体稳定性，能够浓缩 pDNA，编码增强型绿色荧光蛋白和肿瘤抑制因子 p53 基因，形成具有良好细胞相容性的聚阳离子。引人注目的是，在相同条件下，与单 G3 和 G5 PAMAM 树枝状大分子的相应材料相比，所创建的 NC/pDNA 聚阳离子能够将基因转染到癌细胞中的效率分别提高 2.3 倍和 2.1 倍。此外，聚阳离子处理的癌细胞上调了 p53 和 p21 蛋白和 mRNA 的表达水平，并下调了 Cyclin-D1 和 CDK-4 蛋白和 mRNA 的表达，从而将细胞周期阻滞在 G1 期，实现癌症细胞的基因治疗。通过无全身毒性的肿瘤治疗进一步验证了聚阳离子的基因传递效率。该合成的具有低细胞毒性的高效基于树枝状大分子纳米簇的载体系统可能会扩展到解决与遗传障碍相关的各种类型的疾病。
