Serum-Independent Nonviral Gene Delivery to Innate and Adaptive Immune Cells Using Immunoplexes.
Serum-Independent Nonviral Gene Delivery to Innate and Adaptive Immune Cells Using Immunoplexes.
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DOI：
10.1021/acsabm.0c00761
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发表时间：
2020-09-21
影响因子：
4.7
通讯作者：
Pearson RM
中科院分区：
文献类型：
--
作者：
Chakraborty A;Lasola JJM;Truong N;Pearson RM
先天性和适应性免疫细胞的基因工程代表了治疗许多免疫介导的病理的潜在解决方案。目前将核酸高效导入细胞的免疫工程方法依赖于物理机制，如电穿孔、病毒载体或其他化学方法。使用非病毒纳米颗粒的基因递送在生物材料设计中提供了显著的灵活性，以调整关键参数，例如纳米生物相互作用、转染效率和毒性特征。然而，由于复杂的合成程序、在增加的聚合物（氮，N）与DNA比率（磷酸盐，P）（N/P比率）下的高毒性、在血清存在下的差的转染效率和纳米颗粒稳定性以及短期基因表达，它们的临床实用性受到限制。在这里，我们描述了一个简单的，基于聚合物的非病毒基因传递平台的基础上简单的修改聚乙烯亚胺（PEI），显示强大的和血清非依赖性转染的先天性和适应性免疫细胞的发展。合成了阳离子乙酰化PEI（Ac-PEI），并将其与质粒DNA（pDNA）复合，然后用聚（乙烯-alt-马来酸）（PEMA）阴离子包被层包裹，形成免疫复合物（IPs）。通过改变PEMA表达水平，可以在鼠RAW 264.7巨噬细胞、鼠DC 2.4树突状细胞和人Jurkat T细胞中精确控制细胞相互作用和基因表达，从而提供了将特异性细胞靶向工程化到IP平台中的策略。与无包膜对照相比，在血清存在下用于免疫细胞转染的最佳配制的IP利用高N/P比以实现高稳定性，显示出降低的毒性、高基因表达和延长的基因表达持续时间（>3天）。这些结果证明了工程化IP作为简单、模块化、可靶向和有效的非病毒基因递送平台的潜力，以有效地改变免疫系统细胞内的基因表达。
Genetic engineering of innate and adaptive immune cells represents a potential solution to treat numerous immune-mediated pathologies. Current immune engineering methods to introduce nucleic acids into cells with high efficiency rely on physical mechanisms such as electroporation, viral vectors, or other chemical methods. Gene delivery using non-viral nanoparticles offers significant flexibility in biomaterial design to tune critical parameters such as nano-bio interactions, transfection efficiency, and toxicity profiles. However, their clinical utility has been limited due to complex synthetic procedures, high toxicity at increased polymer (nitrogen, N) to DNA ratios (phosphate, P) (N/P ratios), poor transfection efficiency and nanoparticle stability in the presence of serum, and short-term gene expression. Here, we describe the development of a simple, polymer-based non-viral gene delivery platform based on simple modifications of polyethylenimine (PEI) that displays potent and serum-independent transfection of innate and adaptive immune cells. Cationic acetylated PEI (Ac-PEI) was synthesized and complexed with plasmid DNA (pDNA) followed by enveloping with an anionic polyelectrolyte layer of poly(ethylene-alt-maleic acid) (PEMA) to form immunoplexes (IPs). Cellular interactions and gene expression could be precisely controlled in murine RAW 264.7 macrophages, murine DC2.4 dendritic cells, and human Jurkat T cells by altering the levels of PEMA envelopment, thus providing a strategy to engineer specific cell targeting into the IP platform. Optimally formulated IPs for immune cell transfection in the presence of serum utilized high N/P ratios to enable high stability, displayed reduced toxicity, high gene expression, and a lengthened duration of gene expression (>3 days) compared to non-enveloped controls. These results demonstrate the potential of engineered IPs to serve as simple, modular, targetable, and efficient non-viral gene delivery platform to efficiently alter gene expression within cells of the immune system.