Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Biomaterials. 2016 Dec;109:78-87. doi: 10.1016/j.biomaterials.2016.09.006. Epub 2016 Sep 25.
mRNA has broad potential for treating diseases requiring protein expression. However, mRNA can also induce an immune response with associated toxicity. Replacement of uridine bases with pseudouridine has been postulated to modulate both mRNA immunogenicity and potency. Here, we explore the immune response and activity of lipid nanoparticle-formulated unmodified and pseudouridine-modified mRNAs administered systemically in vivo. Pseudouridine modification to mRNA had no significant effect on lipid nanoparticle physical properties, protein expression in vivo, or mRNA immunogenicity compared to unmodified mRNA when delivered systemically with liver-targeting lipid nanoparticles, but reduced in vitro transfection levels. Indicators of a transient, extracellular innate immune response to mRNA were observed, including neutrophilia, myeloid cell activation, and up-regulation of four serum cytokines. This study provides insight into the immune responses to mRNA lipid nanoparticles, and suggests that pseudouridine modifications may be unnecessary for therapeutic application of mRNA in the liver.
mRNA 在治疗需要蛋白质表达的疾病方面具有广泛的潜力。然而，mRNA 也可能引起免疫反应和相关毒性。用假尿嘧啶替代尿嘧啶碱基被认为可以调节 mRNA 的免疫原性和效力。在这里，我们研究了全身给药的未修饰和假尿嘧啶修饰的 mRNA 的脂质纳米颗粒制剂的免疫反应和活性。与未修饰的 mRNA 相比，当用肝靶向脂质纳米颗粒全身给药时，mRNA 的假尿嘧啶修饰对脂质纳米颗粒的物理性质、体内蛋白质表达或 mRNA 免疫原性没有显著影响，但降低了体外转染水平。观察到对外源 mRNA 的短暂的细胞外先天免疫反应的指标，包括中性粒细胞增多、髓样细胞激活和四种血清细胞因子的上调。本研究深入了解了 mRNA 脂质纳米颗粒的免疫反应，并表明在肝脏中应用 mRNA 进行治疗时，假尿嘧啶修饰可能不是必需的。
