Construction of in vivo mRNA delivery systems

项目摘要 细胞靶向给药是药物传递领域的重要研究方向之一。 特别地，对于体内mRNA递送存在巨大的挑战。例如，治疗窗口 目前的输送系统相对狭窄。大量的细胞类型不能在体内有效地递送。 递送材料的生物降解性仍然是一个问题。为了应对挑战，我们的目标是 研究计划是：1）开发不同的脂质衍生物; 2）构建mRNA递送系统; 3） 在动物模型中检查这些系统的递送效率、药代动力学和安全性。在我们 初步研究，我们开发了功能化的脂质样纳米粒子在体内mRNA的传递和基础 编辑.铅材料能够在血友病A中以正常生理水平产生人因子VIII 小鼠在小鼠中也以低剂量实现了有效的碱基编辑。同时，我们构建了维生素 衍生的脂质纳米颗粒，其能够过继巨噬细胞转移以消除多药耐药（MDR） 小鼠模型中的细菌。此外，我们在其他细胞类型中显示了有希望的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，以提高蛋白质产量。通过对内源基因的综合分析， 通过对UTR的表达和从头设计，我们鉴定了5'和3' UTR的最佳组合，称为 NASAR，其比测试的内源UTR显著更有效。这些初步数据 提供科学基础，以解决基于mRNA的疗法的交付挑战。在这一提议中， 我们提出了四个方向的mRNA体内输送：（1）优化N1，N3，N5-三（2-氨乙基）苯-1，3，5- 三羧酰胺（TT）脂质衍生物用于肝细胞递送;（2）研究维生素脂质衍生物用于 巨噬细胞递送;（3）开发用于干细胞递送的糖脂衍生物;（4）构思新的脂质 用于生殖细胞递送的衍生物。我们将在动物实验中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 提供工具来治疗各种疾病。

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.