Department of Biomedical Engineering, Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Institute for Nanobiotechnology, Johns Hopkins University, Baltimore, MD, USA.
Department of Biomedical Engineering, Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
J Control Release. 2017 Oct 10;263:18-28. doi: 10.1016/j.jconrel.2017.03.384. Epub 2017 Mar 27.
Hepatocellular carcinoma (HCC) is the third most deadly cancer in the US, with a meager 5-year survival rate of <20%. Such unfavorable numbers are closely related to the heterogeneity of the disease and the unsatisfactory therapies currently used to manage patients with invasive HCC. Outside of the clinic, gene therapy research is evolving to overcome the poor responses and toxicity associated with standard treatments. The inadequacy of gene delivery vectors, including poor intracellular delivery and cell specificity, are major barriers in the gene therapy field. Herein, we described a non-viral strategy for effective and cancer-specific DNA delivery to human HCC using biodegradable poly(beta-amino ester) (PBAE) nanoparticles (NPs). Varied PBAE NP formulations were evaluated for transfection efficacy and cytotoxicity to a range of human HCC cells as well as healthy human hepatocytes. To address HCC heterogeneity, nine different sources of human HCC cells were utilized. The polymeric NPs composed of 2-((3-aminopropyl)amino) ethanol end-modified poly(1,5-pentanediol diacrylate-co-3-amino-1-propanol) ('536') at a 25 polymer-to-DNA weight-to-weight ratio led to high transfection efficacy to all of the liver cancer lines, but not to hepatocytes. Each individual HCC line had a significantly higher percentage of exogenous gene expression than the healthy liver cells (P<0.01). Notably, this biodegradable end-modified PBAE gene delivery vector was not cytotoxic and maintained the viability of hepatocytes above 80%. In a HCC/hepatocyte co-culture model, in which cancerous and healthy cells share the same micro-environment, 536 25 w/w NPs specifically transfected cancer cells. PBAE NP administration to a subcutaneous HCC mouse model, established with one of the human lines tested in vitro, confirmed effective DNA transfection in vivo. PBAE-based NPs enabled high and preferential DNA delivery to HCC cells, sparing healthy hepatocytes. These biodegradable and liver cancer-selective NPs are a promising technology to deliver therapeutic genes to liver cancer.
肝细胞癌 (HCC) 是美国第三大致命癌症，5 年生存率<20%。如此不利的数字与疾病的异质性以及目前用于治疗侵袭性 HCC 患者的不理想疗法密切相关。在临床之外，基因治疗研究正在发展，以克服与标准治疗相关的不良反应和毒性。包括细胞内递药不佳和细胞特异性在内的基因传递载体的不足，是基因治疗领域的主要障碍。在此，我们描述了一种使用可生物降解的聚（β-氨基酯）（PBAE）纳米颗粒（NP）将有效且针对癌症的 DNA 递送至人类 HCC 的非病毒策略。对不同的 PBAE NP 制剂进行了评估，以确定其对一系列人类 HCC 细胞以及健康的人类肝细胞的转染功效和细胞毒性。为了解决 HCC 的异质性，使用了 9 种不同来源的人类 HCC 细胞。由 2-((3-氨丙基)氨基)乙醇端修饰的聚(1,5-戊二醇二丙烯酸酯-co-3-氨基-1-丙醇)（'536'）组成的聚合物 NP，在 25 聚合物与 DNA 的重量比为重量比时，可高效转染所有肝癌系，但对肝细胞无效。每个 HCC 系的外源基因表达百分比均显著高于健康肝细胞（P<0.01）。值得注意的是，这种可生物降解的端修饰 PBAE 基因传递载体无细胞毒性，可使肝细胞的活力保持在 80%以上。在 HCC/肝细胞共培养模型中，癌变和健康细胞共享相同的微环境，536 25 w/w NPs 特异性转染癌细胞。在体外测试的人类系之一建立的皮下 HCC 小鼠模型中，PBAE NP 给药证实了体内有效的 DNA 转染。PBAE 基 NP 能够高效且优先地将 DNA 递送至 HCC 细胞，而不会损伤健康的肝细胞。这些可生物降解和肝癌选择性的 NP 是向肝癌递送治疗基因的有前途的技术。
