Fusion protein engineered exosomes for targeted degradation of specific RNAs in lysosomes: a proof-of-concept study.
Fusion protein engineered exosomes for targeted degradation of specific RNAs in lysosomes: a proof-of-concept study.
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融合蛋白工程外泌体用于溶酶体中特定 RNA 的靶向降解：一项概念验证研究
DOI：
10.1080/20013078.2020.1816710
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发表时间：
2020-09-06
影响因子：
16
通讯作者：
Yuan L
中科院分区：
文献类型：
--
作者：
Li Z;Zhou X;Gao X;Bai D;Dong Y;Sun W;Zhao L;Wei M;Yang X;Yang G;Yuan L
摘要 治疗性干预体内 RNA 的功能仍然是一个巨大的挑战。我们在这里开发了一种基于外泌体的策略来传递工程化的RNA结合蛋白，以便将特定的RNA招募到溶酶体进行降解。作为一项原理验证研究，RNA 结合蛋白 HuR 融合到 Lamp2b（一种位于外泌体和溶酶体中的膜蛋白）的 C 末端。融合蛋白能够掺入外泌体中。此外，用 Lamp2b-HuR 改造的外泌体可能通过溶酶体介导的降解成功地降低了 RNA 靶标的丰度，特别是当外泌体被酸化时。该系统对巨噬细胞特别有效，巨噬细胞富含溶酶体并且对常规转染介导的 RNAi 策略具有抵抗力。在 CCl4 诱导的肝损伤小鼠模型中，我们发现用 Lamp2b-HuR 工程化的酸化外泌体的递送显着减少了肝纤维化，同时减少了 miR-155 和其他炎症基因。综上所述，所建立的基于外泌体的RNA结合蛋白递送策略，即“外泌体介导的溶酶体清除”，利用了外泌体在靶向递送方面的优势，在体内调控一组基因方面具有广阔的前景。
ABSTRACT Therapeutically intervening the function of RNA in vivo remains a big challenge. We here developed an exosome-based strategy to deliver engineered RNA-binding protein for the purpose of recruiting specific RNA to the lysosomes for degradation. As a proof-of-principle study, RNA-binding protein HuR was fused to the C-terminus of Lamp2b, a membrane protein localized in both exosome and lysosome. The fusion protein was able to be incorporated into the exosomes. Moreover, exosomes engineered with Lamp2b-HuR successfully decreased the abundance of RNA targets possibly via lysosome-mediated degradation, especially when the exosomes were acidified. The system was specifically effective in macrophages, which are lysosome enriched and resistant to routine transfection mediated RNAi strategy. In the CCl4-induced liver injury mouse model, we found that delivery of acidified exosomes engineered with Lamp2b-HuR significantly reduced liver fibrosis, together with decreased miR-155 and other inflammatory genes. In summary, the established exosome-based RNA-binding protein delivery strategy, namely “exosome-mediated lysosomal clearance”, takes the advantage of exosome in targeted delivery and holds great promise in regulating a set of genes in vivo.
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