Construction of in vivo mRNA delivery systems

项目概要 细胞特异性药物递送是药物递送领域最重要的研究领域之一。 特别是，体内 mRNA 递送面临着巨大的挑战。例如，治疗窗 目前的输送系统相对狭窄。许多细胞类型无法在体内有效递送。 输送材料的生物降解性仍然是一个问题。为了应对挑战，我们的目标 研究计划为：1）开发多种脂质衍生物； 2）构建mRNA递送系统； 3）到 检查这些系统在动物模型中的递送效率、药代动力学和安全性。在我们的 初步研究，我们开发了用于体内 mRNA 传递和碱基的功能化脂质样纳米粒子 编辑。该先导材料能够在血友病 A 中产生正常生理水平的人因子 VIII 老鼠。在小鼠体内低剂量下也实现了有效的碱基编辑。同时，我们构建了维生素 衍生的脂质纳米粒子，使巨噬细胞过继转移能够消除多重耐药性（MDR） 小鼠模型中的细菌。此外，我们在其他细胞类型中展示了有希望的 mRNA 递送，例如干细胞 细胞和生殖细胞。此外，我们系统地研究了非翻译区（UTR） mRNA 以增强蛋白质生产。通过内源基因综合分析 通过 UTR 的表达和从头设计，我们确定了 5' 和 3' UTR 的最佳组合，称为 NASAR，其效率明显高于测试的内源 UTR。这些初步数据 为解决基于 mRNA 的疗法的递送挑战提供科学基础。在这个提案中， 我们提出了体内mRNA递送的四个方向：（1）优化N1,N3,N5-三(2-氨乙基)苯-1,3,5- 用于肝细胞递送的三甲酰胺（TT）脂质衍生物； (2) 维生素脂质衍生物的研究 巨噬细胞递送； (3) 开发用于干细胞递送的糖脂衍生物； (4)构想新型脂质 用于生殖细胞递送的衍生物。我们将在动物身上证明细胞特异性递送系统的概念 模型。我们的研究目标是将这一研究策略的创新转化为开发更好的mRNA 治疗多种疾病的输送工具。

Project Summary Cell-specific drug delivery represents one of the most important research areas in the field of drug delivery. Particularly, there are formidable challenges for in vivo mRNA delivery. For example, therapeutic window for current delivery systems is relatively narrow. A large number of cell types cannot be efficiently delivered in vivo. Biodegradability of the delivery materials remains a concern. In order to address the challenges, the goals of our research program are: 1) to develop diverse lipid derivatives; 2) to construct mRNA delivery systems; 3) to examine the delivery efficiency, pharmacokinetics, and safety profile of these systems in animal models. In our preliminary studies, we developed functionalized lipid-like nanoparticles for in vivo mRNA delivery and base editing. The lead material was able to produce human factor VIII at a normal physiological level in hemophilia A mice. The effective base editing was also achieved at low doses in mice. Meanwhile, we constructed vitamin derived lipid nanoparticles, which enabled adoptive macrophage transfer for eliminating multidrug resistant (MDR) bacteria in mouse models. Moreover, we showed promising mRNA delivery in other cell types, such as stem cells and reproductive cells. Additionally, we systematically investigated the untranslated regions (UTRs) of mRNAs in order to enhance protein production. Through a comprehensive analysis of endogenous gene expression and de novo design of UTRs, we identified an optimal combination of 5’ and 3’ UTR, termed as NASAR, which was significantly more efficient than the tested endogenous UTRs. These preliminary data provide the scientific foundation to address the delivery challenges of mRNA-based therapeutics. In this proposal, we propose four directions for mRNA delivery in vivo: (1) to optimize N1,N3,N5-tris(2-aminoethyl)benzene-1,3,5- tricarboxamide (TT) lipid derivatives for hepatocytes delivery; (2) to investigate vitamin lipid derivatives for macrophages delivery; (3) to develop glycolipid derivatives for stem cells delivery; (4) to conceive novel lipid derivatives for reproductive cells delivery. We will prove the concept of cell-specific delivery systems in animal models. Our research goal is to translate the innovations of this research strategy to develop better mRNA delivery tools to treat diverse diseases.