Boosting Intracellular Delivery of Lipid Nanoparticle-Encapsulated mRNA.
Boosting Intracellular Delivery of Lipid Nanoparticle-Encapsulated mRNA.
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DOI：
10.1021/acs.nanolett.7b02664
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发表时间：
2017-09-13
期刊：
影响因子：
10.8
通讯作者：
Sahay G
中科院分区：
文献类型：
--
作者：
Patel S;Ashwanikumar N;Robinson E;DuRoss A;Sun C;Murphy-Benenato KE;Mihai C;Almarsson Ö;Sahay G
信使核糖核酸的细胞内递送在疫苗和治疗的发现和发展方面具有巨大的潜力。尽管基于脂质的纳米颗粒(LNPs)在细胞内传递信使核糖核酸的效用越来越被认识到，但由于对内体逃逸的了解不足，粒子工程受到了阻碍，因为内体逃逸被认为是细胞内核酸在细胞内的可利用性和活性的主要限制因素。利用一系列基于CRISPR的溶酶体途径的遗传扰动，我们已经确认晚期内小体/溶酶体(LE/Ly)的形成对于外源呈现的mRNA的功能递送是必不可少的。溶酶体提供了一个时空枢纽来协调mTOR信号，并被认为控制细胞增殖、营养感知、核糖体生物发生和mRNA翻译。通过调节mTOR通路，我们能够增强或抑制LNP介导的mRNA传递。为了进一步促进mRNA在细胞内的传递，我们筛选了212个生物活性类脂分子，这些分子要么富含在囊泡中，要么调节细胞信号。令人惊讶的是，我们发现，临床上被批准用于治疗哮喘和其他肺部疾病的白三烯拮抗剂，在体外(超过3倍，p<；0.005)和体内(超过2倍，p<；0.005)，增加了细胞内的mRNA量。了解LNP介导的细胞内递送将激励下一代具有高效力和。毒性有限。
Intracellular delivery of mRNA holds great potential for vaccine and therapeutic discovery and development. Despite increasing recognition of the utility of lipid-based nanoparticles (LNPs) for intracellular delivery of mRNA, particle engineering is hindered by insufficient understanding of endosomal escape, which is believed to be a main limiter of cytosolic availability and activity of the nucleic acid inside the cell. Using a series of CRISPR-based genetic perturbations of the lysosomal pathway, we have identified that late endosome/lysosome (LE/Ly) formation is essential for functional delivery of exogenously presented mRNA. Lysosomes provide a spatio-temporal hub to orchestrate mTOR signaling and are known to control cell proliferation, nutrient sensing, ribosomal biogenesis, and mRNA translation. Through modulation of the mTOR pathway we were able to enhance or inhibit LNP-mediated mRNA delivery. To further boost intracellular delivery of mRNA we screened 212 bioactive lipid-like molecules that are either enriched in vesicular compartments or modulate cell signaling. Surprisingly, we have discovered that leukotriene-antagonists, clinically approved for treatment of asthma and other lung diseases, enhance intracellular mRNA delivery in vitro (over 3-fold, p<0.005) and in vivo (over 2-fold, p<0.005). Understanding LNP-mediated intracellular delivery will inspire the next generation of RNA therapeutics that have high potency and. limited toxicity.
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影响因子：
5.1
作者：
DeRosa, F.;Guild, B.;Karve, S.;Smith, L.;Love, K.;Dorkin, J. R.;Kauffman, K. J.;Zhang, J.;Yahalom, B.;Anderson, D. G.;Heartlein, M. W.
通讯作者：
Heartlein, M. W.
DOI：
10.1126/science.aag1417
发表时间：
2017-03-24
期刊：
Science (New York, N.Y.)
影响因子：
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通讯作者：
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影响因子：
46.9
作者：
Gilleron, Jerome;Querbes, William;Zerial, Marino
通讯作者：
Zerial, Marino
影响因子：
15
作者：
Chen, Delai;Love, Kevin T.;Anderson, Daniel G.
通讯作者：
Anderson, Daniel G.
DOI：
10.1083/jcb.201507112
发表时间：
2016-03-14
期刊：
The Journal of cell biology
影响因子：
--
作者：
Johnson DE;Ostrowski P;Jaumouillé V;Grinstein S
通讯作者：
Grinstein S