Mohri Kohta, Miyazaki Takayuki, Warashina Shota, Takahashi Maiko, Ren Qin, Iida Riho, Wada Yasuhiro, Maeda Kazuya, Watanabe Yasuyoshi, Suzuki Yuta, Mukai Hidefumi
Laboratory for Molecular Delivery and Imaging Technology, RIKEN Center for Biosystems Dynamics Research, 6-7-3 minatojima-minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan.
Department of Pharmaceutical Informatics, Graduate School of Biomedical Science, Nagasaki University, 1-7-1 Sakamoto, Nagasaki-shi, Nagasaki 852-8588, Japan.
ACS Appl Mater Interfaces. 2025 Aug 13;17(32):45625-45639. doi: 10.1021/acsami.5c14143. Epub 2025 Aug 3.
It is important to clarify the pharmacokinetics of mRNA-loaded lipid nanoparticles (mRNA-LNPs), which have attracted attention as new pharmaceutical modalities for vaccines and therapeutic agents against various diseases. Positron emission tomography (PET) is expected to provide data to ensure the pharmacokinetics of mRNA-LNPs early after administration in detail, especially because of its high temporal resolution. In this study, we have developed a method for the preparation of Cu-labeled mRNA-LNPs by an approach via hybridization of short oligonucleotides to the 3' UTR region of mRNA, and conducted PET pharmacokinetic studies in normal and functionally deficient mice after intravenous and intramuscular administration, which mainly yielded the following findings. First, the common composition of mRNA-LNPs showed a clearly separated two-step accumulation process in the liver after intravenous administration in normal mice, indicating the involvement of ApoE in the second step. Second, after intravenous administration, the time to protein corona formation, in which blood proteins bind to mRNA-LNPs, was found to be within 5-10 min. Third, liver and spleen clearance was determined using integration plot analysis, and the contribution of hepatocyte uptake via ApoE to hepatic clearance after intravenous administration was approximately twice that of Kupffer cell uptake. Fourth, a comparison with luciferase reporter protein expression suggested that the protein expression ability of hepatocytes taken up via ApoE after intravenous administration and that of antigen-presenting cells transferred to the lymph nodes after intramuscular administration was remarkably high.
阐明负载信使核糖核酸的脂质纳米颗粒（mRNA-LNPs）的药代动力学很重要，mRNA-LNPs作为针对各种疾病的疫苗和治疗剂的新型药物形式已引起关注。正电子发射断层扫描（PET）有望提供数据，以便在给药后早期详细确定mRNA-LNPs的药代动力学，特别是由于其具有高时间分辨率。在本研究中，我们开发了一种通过短寡核苷酸与mRNA的3'UTR区域杂交的方法来制备铜标记的mRNA-LNPs，并在正常和功能缺陷小鼠经静脉和肌肉注射后进行了PET药代动力学研究，主要得出以下结果。首先，mRNA-LNPs的常见组成在正常小鼠静脉注射后在肝脏中显示出明显分离的两步积累过程，表明载脂蛋白E（ApoE）参与第二步。其次，静脉注射后，发现血液蛋白与mRNA-LNPs结合形成蛋白冠的时间在5-10分钟内。第三，使用积分图分析确定肝脏和脾脏清除率，静脉注射后通过ApoE的肝细胞摄取对肝脏清除的贡献约为库普弗细胞摄取的两倍。第四，与荧光素酶报告蛋白表达的比较表明，静脉注射后通过ApoE摄取的肝细胞以及肌肉注射后转移至淋巴结的抗原呈递细胞的蛋白表达能力非常高。
