Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
J Biomed Mater Res A. 2013 Jul;101(7):1837-45. doi: 10.1002/jbm.a.34616. Epub 2013 Apr 5.
Liver cancer is a leading cause of cancer death. Most patients are treated by arterial injection of chemoembolizing agents, providing a convenient avenue for local treatment by novel therapies, including gene therapy. Poly(beta-amino ester)s (PBAEs) were synthesized and used to form nanoparticles for non-viral transfection of buffalo rat hepatoma (MCA-RH7777) and hepatocyte (BRL-3A) lines with eGFP and luciferase DNA. Hepatoma cells were transfected with up to (98 ± 0.4)% efficacy with no measurable cytotoxicity. Hepatocytes were transfected with as high as (73 ± 0.4)% efficacy with (10 ± 4)% non-specific cytotoxicity. In contrast, positive controls (branched polyethylenimine, Lipofectamine™ 2000, and X-tremeGENE(®) DNA HP) caused 30-90% toxicity in BRL-3A cells at doses required for >50% transfection. Of the 21 optimized PBAE-DNA formulations tested, 12 showed significant specificity for hepatoma cells over hepatocytes in monoculture (p < 0.05 for both percentage transfected and eGFP expression intensity). Top polymers from eGFP studies also delivered luciferase DNA with 220 ± 30-fold and 470 ± 30-fold greater specificity for hepatoma cells than hepatocytes. Transfections of co-cultures of hepatoma and hepatocytes with eGFP DNA also showed high specificity (1.9 ± 0.1- to 5.8± 1.4-fold more transfected hepatoma cells than hepatocytes, measured by percentage transfected and flow cytometry). By eGFP intensity, up to 530 ±60-fold higher average expression per cell was measured in hepatoma cells. One top formulation caused (95 ± 0.2)% transfection in hepatoma cells and (27 ± 0.2)% in hepatocytes [(96 ± 9)% relative hepatocyte viability]. PBAE-based nanoparticles are a viable strategy for liver cancer treatment, delivering genes to nearly 100% of cancer cells while maintaining high biomaterial-mediated specificity to prevent toxic side-effects on healthy hepatocytes.
肝癌是癌症死亡的主要原因。大多数患者通过动脉内注射化疗栓塞剂进行治疗，这为包括基因治疗在内的新型疗法提供了局部治疗的便利途径。我们合成了聚（β-氨基酯）（PBAE）并将其用于形成纳米颗粒，用于水牛大鼠肝癌（MCA-RH7777）和肝细胞（BRL-3A）系的非病毒转染，转染带有 eGFP 和荧光素酶 DNA。肝癌细胞的转染效率高达（98±0.4）%，且没有可测量的细胞毒性。转染肝细胞的效率高达（73±0.4）%，同时仅有（10±4）%的非特异性细胞毒性。相比之下，阳性对照（支化聚乙烯亚胺、LipofectamineTM 2000 和 X-tremeGENE(®) DNA HP）在需要 >50%转染的剂量下会导致 BRL-3A 细胞 30-90%的毒性。在测试的 21 种优化的 PBAE-DNA 制剂中，有 12 种在单核培养中对肝癌细胞具有明显的特异性，明显高于对肝细胞的特异性（转染百分比和 eGFP 表达强度的 p 值均 <0.05）。来自 eGFP 研究的最佳聚合物也能以 220±30 倍和 470±30 倍的特异性将荧光素酶 DNA 递送给肝癌细胞，高于肝细胞。共培养肝癌细胞和肝细胞的 eGFP DNA 转染也显示出高特异性（通过转染百分比和流式细胞术测量，转染的肝癌细胞比肝细胞多 1.9±0.1-5.8±1.4 倍）。通过 eGFP 强度，每个细胞的平均表达量高达 530±60 倍。一种最佳制剂在肝癌细胞中的转染效率为（95±0.2）%，在肝细胞中的转染效率为（27±0.2）%[（96±9）%相对肝细胞活力]。基于 PBAE 的纳米颗粒是治疗肝癌的一种可行策略，能将近 100%的癌细胞转染，同时保持对健康肝细胞的高生物材料介导的特异性，以防止对其产生毒性副作用。
