An optimized exosome production strategy for enhanced yield while without sacrificing cargo loading efficiency.
An optimized exosome production strategy for enhanced yield while without sacrificing cargo loading efficiency.
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DOI：
10.1186/s12951-022-01668-3
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发表时间：
2022-10-29
影响因子：
10.2
通讯作者：
Yang, Guodong
中科院分区：
文献类型：
--
作者：
Zhang, Rongxin;Bu, Te;Cao, Ruidan;Li, Zhelong;Wang, Chen;Huang, Bing;Wei, Mengying;Yuan, Lijun;Yang, Guodong
外泌体介导的mRNA递送是用于治疗多种疾病的有前景的策略。然而，外泌体的低产量是临床翻译的瓶颈。在这项研究中，我们通过同时减少抑制外泌体生物发生的基因的表达和向培养基中补充红细胞膜成分来促进外泌体的产生。在候选基因中，Rab 4的敲除被鉴定为在促进外来体生物发生方面具有最高功效，同时没有任何明显的细胞毒性。此外，在培养基中补充红细胞膜颗粒（RCMP）进一步促进外泌体产生。Rab 4敲除和RCMP补充的组合使外泌体产量增加高达14倍。作为概念验证研究，低密度脂蛋白受体（Ldlr）mRNA在外泌体供体细胞中强制表达，并在生物发生过程中被动封装到外泌体中。虽然每个细胞的外泌体产量增加，但加强策略没有改变每个外泌体的治疗性Ldlr mRNA的装载效率。同样，通过该策略衍生的治疗性外泌体减轻了Ldlr−/−小鼠的肝脏脂肪变性和动脉粥样硬化，与常规方法产生的外泌体相似。总之，所提出的外泌体助推器策略在一定程度上克服了低产率瓶颈，并且肯定会促进外泌体的临床翻译。在线版本包含补充材料，可通过10.1186/s12951-022-01668-3获得。
Exosome mediated mRNA delivery is a promising strategy for the treatment of multiple diseases. However, the low yield of exosomes is a bottleneck for clinical translation. In this study, we boosted exosome production via simultaneously reducing the expression of genes inhibiting exosome biogenesis and supplementing the culture medium with red cell membrane components. Among the candidate genes, knocking down of Rab4 was identified to have the highest efficacy in promoting exosome biogenesis while without any obvious cytotoxicity. Additionally, supplementing red cell membrane particles (RCMPs) in the culture medium further promoted exosome production. Combination of Rab4 knockdown and RCMP supplement increased exosome yield up to 14-fold. As a proof-of-concept study, low-density lipoprotein receptor (Ldlr) mRNA was forced expressed in the exosome donor cells and passively encapsulated into the exosomes during biogenesis with this strategy. Though exosome production per cell increased, the booster strategy didn’t alter the loading efficiency of therapeutic Ldlr mRNA per exosome. Consistently, the therapeutic exosomes derived by the strategy alleviated liver steatosis and atherosclerosis in Ldlr−/− mice, similar as the exosomes produced by routine methods. Together, the proposed exosome booster strategy conquers the low yield bottleneck to some extent and would certainly facilitate the clinical translation of exosomes. The online version contains supplementary material available at 10.1186/s12951-022-01668-3.
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DOI：
10.1097/shk.0000000000000995
发表时间：
2018-03
期刊：
Shock (Augusta, Ga.)
影响因子：
--
作者：
Kim Y;Abplanalp WA;Jung AD;Schuster RM;Lentsch AB;Gulbins E;Caldwell CC;Pritts TA
通讯作者：
Pritts TA
影响因子：
46.9
作者：
Alvarez-Erviti, Lydia;Seow, Yiqi;Wood, Matthew J. A.
通讯作者：
Wood, Matthew J. A.
DOI：
10.1126/science.aau6977
发表时间：
2020-02-07
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Kalluri R;LeBleu VS
通讯作者：
LeBleu VS
DOI：
10.1073/pnas.1200448109
发表时间：
2012-03-13
影响因子：
11.1
作者：
Nabhan, Joseph F.;Hu, Ruoxi;Lu, Quan
通讯作者：
Lu, Quan
影响因子：
4
作者：
Colombo, Marina;Moita, Catarina;Raposo, Graca
通讯作者：
Raposo, Graca