Construction of in vivo mRNA delivery systems

项目摘要 细胞特异性药物传递是药物传递领域中最重要的研究领域之一。 特别是，体内信使核糖核酸的传递面临着巨大的挑战。例如，治疗窗口期 目前的投放系统相对较窄。大量的细胞类型不能在体内有效地传递。 输送材料的生物降解性仍然是一个令人担忧的问题。为了应对挑战，我们的目标是 研究项目有：1)开发多种脂类衍生物；2)构建信使核糖核酸递送系统；3) 在动物模型中检查这些系统的给药效率、药代动力学和安全性。在我们的 初步研究，我们开发了功能化的类脂纳米粒用于体内信使核糖核酸的传递和碱基 正在编辑。在血友病A中，铅材料能够在正常生理水平下产生人类第八因子 老鼠。在小鼠体内也能在低剂量下实现有效的碱基编辑。同时，我们制造了维他命 衍生脂质纳米粒，使过继巨噬细胞转移消除多药耐药(MDR)成为可能 小鼠模型中的细菌。此外，我们还展示了在其他类型的细胞中有希望的信使核糖核酸的传递，例如干细胞 细胞和生殖细胞。此外，我们还系统地研究了未翻译区域(UTRs) MRNAs，以提高蛋白质产量。通过对内源基因的综合分析 表达和从头设计，我们确定了5‘和3’非翻译区的最佳组合，称为 NASAR，其效率显著高于所测试的内源UTRs。这些初步数据 为解决基于信使核糖核酸的疗法的传递挑战提供科学基础。在这份提案中， 我们提出了四个研究方向：(1)优化N_1，N_3，N_5-三(2-氨基乙基)苯-1，3，5- 用于肝细胞转运的三羧胺(TT)脂类衍生物；(2)研究维生素脂类衍生物 巨噬细胞转运；(3)开发干细胞输送用糖脂衍生物；(4)构思新脂质 用于生殖细胞输送的衍生品。我们将在动物身上证明细胞特异性递送系统的概念 模特们。我们的研究目标是将这一研究策略的创新转化为更好的mRNA 治疗各种疾病的递送工具。

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.