The demand for RNA-based therapeutics is increasing globally. However, their use is hampered by the lack of safe and effective delivery vehicles. Here, we developed technologies for highly efficient delivery of RNA cargo into programmable extracellular vesicle-mimetic nanovesicles (EMNVs) by fabricating hybrid EMNV-liposomes (Hybs). Tissue targeting is endowed by highly efficient genetic platforms based on truncated CD63 (ΔCD63) or PTGFRN proteins. For the first time we reveal their efficiency in functionalizing EMNVs, resulting in >10-fold enhancement of nanoparticle internalization in vitro and >2-fold in vivo. RNA delivery using Hybs demonstrated efficiency of >85% in human and mouse cell lines. Comparative analysis of EMNVs and Hyb lysosome colocalization and stability suggested that Hybs enter the lysosomal compartment and escape over time, whereas EMNVs primarily avoid it. Finally, we used these technologies to generate liver-targeting Hybs loaded with therapeutic small interfering RNA and demonstrated the robust efficiency of this system in vitro and in vivo. These technologies can be adapted for manufacturing a wide range of next-generation vehicles for highly efficient, safe delivery of RNA into desired organs and tissues for therapeutic and prophylactic applications.

中文翻译：

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基于生物制剂的技术，用于高效和靶向 RNA 递送


全球对基于 RNA 的疗法的需求正在增加。然而，由于缺乏安全有效的送货车辆，它们的使用受到了阻碍。在这里，我们开发了通过制造杂交 EMNV-脂质体 （Hybs） 将 RNA 货物高效递送到可编程细胞外囊泡模拟纳米囊泡 （EMNV） 中的技术。组织靶向由基于截短 CD63 （ΔCD63） 或 PTGFRN 蛋白的高效遗传平台赋予。我们首次揭示了它们在功能化 EMNV 方面的效率，导致纳米颗粒在体外内化 的 >10 倍增强，在 体内 >2 倍。使用 Hybs 的 RNA 递送在人和小鼠细胞系中显示出 >85% 的效率。EMNVs 和 Hyb 溶酶体共定位和稳定性的比较分析表明，Hybs 进入溶酶体区室并随着时间的推移而逃逸，而 EMNVs 主要避开它。最后，我们使用这些技术生成了载有治疗性小干扰 RNA 的肝脏靶向 Hybs，并在体外 和体内证明了该系统 的稳健效率。这些技术可用于制造各种下一代载体，以高效、安全地将 RNA 递送到所需的器官和组织中，用于治疗和预防应用。