Impact of mRNA chemistry and manufacturing process on innate immune activation
Impact of mRNA chemistry and manufacturing process on innate immune activation
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DOI：
10.1126/sciadv.aaz6893
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发表时间：
2020-06-01
期刊：
影响因子：
13.6
通讯作者：
Joyal, John L.
中科院分区：
文献类型：
--
作者：
Nelson, Jennifer;Sorensen, Elizabeth W.;Joyal, John L.
信使RNA（mRNA）代表了潜在广泛的临床适应症的有吸引力的治疗方式，但需要尿苷化学修饰和/或调节生产过程以防止细胞先天免疫传感器的激活和伴随的蛋白质表达的减少。为了破译这些因素对免疫激活的相对贡献，在这里，我们比较了，在多个细胞和体内模型，mRNA编码的人促红细胞生成素纳入规范尿苷或N1-甲基-假尿苷（1 m Psi），合成的标准过程中显示有双链RNA（dsRNA）杂质或修改的过程，产生高度纯化的mRNA制剂。我们的数据表明，免疫终点的最低刺激是由修改后的方法制成的1 m Psi，而含有规范尿苷的mRNA是免疫刺激性的，无论过程如何。这些发现证实，尿苷修饰和dsRNA杂质的减少在控制治疗性mRNA的免疫激活谱方面是必要的和足够的。
Messenger RNA (mRNA) represents an attractive therapeutic modality for potentially a wide range of clinical indications but requires uridine chemistry modification and/or tuning of the production process to prevent activation of cellular innate immune sensors and a concomitant reduction in protein expression. To decipher the relative contributions of these factors on immune activation, here, we compared, in multiple cell and in vivo models, mRNA that encodes human erythropoietin incorporating either canonical uridine or N1-methyl-pseudouridine (1 m Psi), synthesized by either a standard process shown to have double-stranded RNA (dsRNA) impurities or a modified process that yields a highly purified mRNA preparation. Our data demonstrate that the lowest stimulation of immune endpoints was with 1 m Psi made by the modified process, while mRNA containing canonical uridine was immunostimulatory regardless of process. These findings confirm that uridine modification and the reduction of dsRNA impurities are both necessary and sufficient at controlling the immune-activating profile of therapeutic mRNA.