拉吉细胞
电穿孔
转染
嵌合抗原受体
Jurkat细胞
T细胞
细胞生物学
细胞培养
细胞
化学
免疫学
基因
免疫系统
生物
遗传学
生物化学
作者
Yaping Chen,Melanie Mach,Ali‐Reza Shokouhi,Hao Zhe Yoh,David Bishop,Takahide Murayama,Koukou Suu,Yasuhiro Morikawa,Simon C. Barry,Kenneth Micklethwaite,Roey Elnathan,Nicolas H. Voelcker
标识
DOI:10.1016/j.mattod.2023.02.009
摘要
Cell-based immunotherapy such as chimeric antigen receptor (CAR)-T therapy holds great promise in treating cancer and other diseases; but the current viral-based method represents a significant cost and safety hurdle. Here, we show for the first time successful CAR transfection into primary T cells via vertically aligned silicon nanotube (SiNT) arrays. SiNT-mediated transfection achieves comparable or even higher delivery efficiency (20–37%) and expression efficiency (18–24%) to that achieved by electroporation. Scanning electron microscopy imaging after focused ion beam milling demonstrated the tight T cell–SiNT interface. The induced membrane invaginations and the proximity between individual SiNTs and the nucleus might enhance endocytic pathways, and enable direct delivery of CAR construct into the nucleus, thus resulting in higher CAR expression efficiency. SiNT-interfacing also results in faster proliferation of T cells compared to cells transfected by electroporation; non-activated T (N_SiNT) cells undergo higher numbers of cell division than pre-activated ones (A_SiNT). By co-culturing with target lymphoma Raji cells, we prove that SiNT-transfected CAR-T cells can suppress Raji cell growth, indicated by significant increase in effector:target (E:T) ratio (by up to 30.7-fold). While SiNTs induce an overall upregulation of cytokine production in T cells, N_SiNT T cells exhibited high increase in secretion of IFNγ and IL-6, and relatively high in TNFα, which could contribute to their enhanced killing ability (∼96% cytotoxicity), demonstrated by their stronger inhibition on target Raji cells through luciferase assay. The results demonstrate the capacity of SiNT-mediated transfection of generating effective anti-lymphoma CAR-T cells. Considering the growing potential of cell-based therapies, we expect that a non-viral nanoinjection platform such as ours will facilitate the full realization of their therapeutic promise.
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