Department of Neurosurgery, Huashan Hospital, Shanghai Medical College, Fudan University, National Center for Neurological Disorders, National Key Laboratory for Medical Neurobiology, Institutes of Brain Science, Shanghai Key Laboratory of Brain Function and Regeneration, Institute of Neurosurgery, MOE Frontiers Center for Brain Science, Shanghai, 200040, China.
Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
Adv Sci (Weinh). 2023 Apr;10(10):e2206517. doi: 10.1002/advs.202206517. Epub 2023 Feb 2.
Engineered extracellular vesicles (EVs) are considered excellent delivery vehicles for a variety of therapeutic agents, including nucleic acids, proteins, drugs, and nanomaterials. Recently, several studies have indicated that clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) delivered by EVs enable efficient DNA editing. However, an RNA editing tool delivered by EVs is still unavailable. Here, a signal peptide-optimized and EVs-delivered guide RNA (gRNA) and CRISPR/CasRx (Cas13d) system capable of rapidly inhibiting the expression of targeted genes with quick catabolism after performing their functions is developed. EVs with CRISPR/CasRx and tandem gRNAs targeting pivotal cytokines are further packed whose levels increase substantially over the course of acute inflammatory diseases and find that these engineered EVs inhibit macrophage activation in vitro. More importantly, this system attenuates lipopolysaccharide (LPS)-triggered acute lung injury and sepsis in the acute phase, mitigating organ damage and improving the prognosis in vivo. In summary, a potent tool is provided for short-acting RNA editing, which could be a powerful therapeutic platform for the treatment of acute diseases.
工程细胞外囊泡（EVs）被认为是各种治疗剂的理想递送载体，包括核酸、蛋白质、药物和纳米材料。最近，几项研究表明，EV 递送的成簇规律间隔短回文重复序列（CRISPR）/CRISPR 相关 9（Cas9）可实现有效的 DNA 编辑。然而，EV 递送的 RNA 编辑工具仍然不可用。在这里，开发了一种信号肽优化的 EV 递送向导 RNA（gRNA）和 CRISPR/CasRx（Cas13d）系统，该系统能够快速抑制靶向基因的表达，并在发挥作用后快速代谢。进一步包装了针对关键细胞因子的 CRISPR/CasRx 和串联 gRNA 的 EV，在急性炎症性疾病过程中其水平显著增加，发现这些工程化的 EV 可抑制体外巨噬细胞的激活。更重要的是，该系统可减轻脂多糖（LPS）引发的急性肺损伤和败血症的急性阶段，减轻器官损伤并改善体内预后。总之，为短期 RNA 编辑提供了一种有效工具，可为急性疾病的治疗提供一种强大的治疗平台。
