阳离子聚合物用于非病毒基因递送至人 T 细胞。
Cationic polymers for non-viral gene delivery to human T cells.
机构信息
Department of Bioengineering and Molecular Engineering and Sciences Institute, University of Washington, Seattle, WA 98195, USA.
Department of Bioengineering and Molecular Engineering and Sciences Institute, University of Washington, Seattle, WA 98195, USA.
出版信息
J Control Release. 2018 Jul 28;282:140-147. doi: 10.1016/j.jconrel.2018.02.043. Epub 2018 Mar 5.
The clinical success of chimeric antigen receptor (CAR) T cell immunotherapy in treating multiple blood cancers has created a need for efficient methods of ex vivo gene delivery to primary human T cells for cell engineering. Here, we synthesize and evaluate a panel of cationic polymers for gene delivery to both cultured and primary human T cells. We show that a subset of comb- and sunflower-shaped pHEMA-g-pDMAEMA polymers can mediate transfection with efficiencies up to 50% in the Jurkat human T cell line with minimal concomitant toxicity (>90% viability). We then optimize primary human T cell transfection conditions including activation time, cell density, DNA dose, culture media, and cytokine treatment. We demonstrate transfection of both CD4 and CD8 primary human T cells with messenger RNA and plasmid DNA at efficiencies up to 25 and 18%, respectively, with similarly high viability.
嵌合抗原受体 (CAR) T 细胞免疫疗法在治疗多种血液癌症方面的临床成功，催生了将基因有效地递送至原代人 T 细胞以进行细胞工程改造的需求。在这里，我们合成并评估了一系列阳离子聚合物，以用于向培养的和原代人 T 细胞进行基因传递。我们发现，一组梳状和向日葵状的 pHEMA-g-pDMAEMA 聚合物可以介导转染，在 Jurkat 人 T 细胞系中的效率高达 50%，同时伴随的毒性最小（>90%的活力）。然后，我们优化了原代人 T 细胞的转染条件，包括激活时间、细胞密度、DNA 剂量、培养基和细胞因子处理。我们证明了信使 RNA 和质粒 DNA 可以分别有效地转染 CD4 和 CD8 原代人 T 细胞，效率高达 25%和 18%，同时具有相似的高活力。
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