Image-based analysis of lipid nanoparticle-mediated siRNA delivery, intracellular trafficking and endosomal escape

被引：1162

作者：

Gilleron, Jerome ^{[1
]}

Querbes, William ^{[2
]}

Zeigerer, Anja ^{[1
]}

Borodovsky, Anna ^{[2
]}

Marsico, Giovanni ^{[1
]}

Schubert, Undine ^{[1
]}

Manygoats, Kevin ^{[1
]}

Seifert, Sarah ^{[1
]}

Andree, Cordula ^{[1
]}

Stoeter, Martin ^{[1
]}

Epstein-Barash, Hila ^{[2
]}

Zhang, Ligang ^{[2
]}

Koteliansky, Victor ^{[2
]}

Fitzgerald, Kevin ^{[2
]}

Fava, Eugenio ^{[1
,3
]}

Bickle, Marc ^{[1
]}

Kalaidzidis, Yannis ^{[1
,4
]}

Akinc, Akin ^{[2
]}

Maier, Martin ^{[2
]}

Zerial, Marino ^{[1
]}

机构：

[1] Max Planck Inst Mol Cell Biol & Genet, Dresden, Germany

[2] Alnylam Pharmaceut, Cambridge, MA USA

[3] German Ctr Neurodegenerat Dis, Bonn, Germany

[4] Moscow MV Lomonosov State Univ, Fac Bioengn & Bioinformat, Moscow, Russia

来源：

NATURE BIOTECHNOLOGY | 2013年 / 31卷 / 07期

关键词：

IN-VIVO; TARGETED DELIVERY; RNAI THERAPEUTICS; CATIONIC LIPIDS; INTERFERING RNA; GENE; MECHANISMS; EFFICIENT; DESIGN; RAB;

D O I：

10.1038/nbt.2612

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Delivery of short interfering RNAs (siRNAs) remains a key challenge in the development of RNA interference (RNAi) therapeutics. A better understanding of the mechanisms of siRNA cellular uptake, intracellular transport and endosomal release could critically contribute to the improvement of delivery methods. Here we monitored the uptake of lipid nanoparticles (LNPs) loaded with traceable siRNAs in different cell types in vitro and in mouse liver by quantitative fluorescence imaging and electron microscopy. We found that LNPs enter cells by both constitutive and inducible pathways in a cell type-specific manner using clathrin-mediated endocytosis as well as macropinocytosis. By directly detecting colloidal-gold particles conjugated to siRNAs, we estimated that escape of siRNAs from endosomes into the cytosol occurs at low efficiency (1-2%) and only during a limited window of time when the LNPs reside in a specific compartment sharing early and late endosomal characteristics. Our results provide insights into LNP-mediated siRNA delivery that can guide development of the next generation of delivery systems for RNAi therapeutics.

引用

页码：638 / U102

页数：12

共 49 条

[1] Modulation of thermal stability can enhance the potency of siRNA [J].

Addepalli, Haripriya ;

Meena ;

Peng, Chang G. ;

Wang, Gang ;

Fan, Yupeng ;

Charisse, Klaus ;

Jayaprakash, K. Narayanannair ;

Rajeev, Kallanthottathil G. ;

Pandey, Rajendra K. ;

Lavine, Gary ;

Zhang, Ligang ;

Jahn-Hofmann, Kerstin ;

Hadwiger, Philipp ;

Manoharan, Muthiah ;

Maier, Martin A. .

NUCLEIC ACIDS RESEARCH, 2010, 38 (20) :7320-7331

[2] A combinatorial library of lipid-like materials for delivery of RNAi therapeutics [J].

Akinc, Akin ;

Zumbuehl, Andreas ;

Goldberg, Michael ;

Leshchiner, Elizaveta S. ;

Busini, Valentina ;

Hossain, Naushad ;

Bacallado, Sergio A. ;

Nguyen, David N. ;

Fuller, Jason ;

Alvarez, Rene ;

Borodovsky, Anna ;

Borland, Todd ;

Constien, Rainer ;

de Fougerolles, Antonin ;

Dorkin, J. Robert ;

Jayaprakash, K. Narayanannair ;

Jayaraman, Muthusamy ;

John, Matthias ;

Koteliansky, Victor ;

Manoharan, Muthiah ;

Nechev, Lubomir ;

Qin, June ;

Racie, Timothy ;

Raitcheva, Denitza ;

Rajeev, Kallanthottathil G. ;

Sah, Dinah W. Y. ;

Soutschek, Juergen ;

Toudjarska, Ivanka ;

Vornlocher, Hans-Peter ;

Zimmermann, Tracy S. ;

Langer, Robert ;

Anderson, Daniel G. .

NATURE BIOTECHNOLOGY, 2008, 26 (05) :561-569

[3] Targeted Delivery of RNAi Therapeutics With Endogenous and Exogenous Ligand-Based Mechanisms [J].

Akinc, Akin ;

Querbes, William ;

De, Soma ;

Qin, June ;

Frank-Kamenetsky, Maria ;

Jayaprakash, K. Narayanannair ;

Jayaraman, Muthusamy ;

Rajeev, Kallanthottathil G. ;

Cantley, William L. ;

Dorkin, J. Robert ;

Butler, James S. ;

Qin, LiuLiang ;

Racie, Timothy ;

Sprague, Andrew ;

Fava, Eugenio ;

Zeigerer, Anja ;

Hope, Michael J. ;

Zerial, Marino ;

Sah, Dinah W. Y. ;

Fitzgerald, Kevin ;

Tracy, Mark A. ;

Manoharan, Muthiah ;

Koteliansky, Victor ;

de Fougerolles, Antonin ;

Maier, Martin A. .

MOLECULAR THERAPY, 2010, 18 (07) :1357-1364

[4] Nanoparticles for ex vivo siRNA delivery to dendritic cells for cancer vaccines: Programmed endosomal escape and dissociation [J].

Akita, Hidetaka ;

Kogure, Kentaro ;

Moriguchi, Rumiko ;

Nakamura, Yoshio ;

Higashi, Tomoko ;

Nakamura, Takashi ;

Serada, Satoshi ;

Fujimoto, Minoru ;

Naka, Tetsuji ;

Futaki, Shiroh ;

Harashima, Hideyoshi .

JOURNAL OF CONTROLLED RELEASE, 2010, 143 (03) :311-317

[5] Design of a peptide-based vector, PepFect6, for efficient delivery of siRNA in cell culture and systemically in vivo [J].

Andaloussi, Samir E. L. ;

Lehto, Taavi ;

Maeger, Imre ;

Rosenthal-Aizman, Katri ;

Oprea, Iulian I. ;

Simonson, Oscar E. ;

Sork, Helena ;

Ezzat, Kariem ;

Copolovici, Dana M. ;

Kurrikoff, Kaido ;

Viola, Joana R. ;

Zaghloul, Eman M. ;

Sillard, Rannar ;

Johansson, Henrik J. ;

Hassane, Fatouma Said ;

Guterstam, Peter ;

Suhorutsenko, Julia ;

Moreno, Pedro M. D. ;

Oskolkov, Nikita ;

Haelldin, Jonas ;

Tedebark, Ulf ;

Metspalu, Andres ;

Lebleu, Bernard ;

Lehtioe, Janne ;

Smith, C. I. Edvard ;

Langel, Uelo .

NUCLEIC ACIDS RESEARCH, 2011, 39 (09) :3972-3987

[6]

[Anonymous], 2011, NCT01437059, Patent No. [7745608, 7,745,608]

[7] Effective siRNA delivery and target mRNA degradation using an amphipathic peptide to facilitate pH-dependent endosomal escape [J].

Bartz, Rene ;

Fan, Haihong ;

Zhang, Jingtao ;

Innocent, Nathalie ;

Cherrin, Craig ;

Beck, Stephen C. ;

Pei, Yi ;

Momose, Aaron ;

Jadhav, Vasant ;

Tellers, David M. ;

Meng, Fanyu ;

Crocker, Louis S. ;

Sepp-Lorenzino, Laura ;

Barnett, Stanley F. .

BIOCHEMICAL JOURNAL, 2011, 435 :475-487

[8] Influence of Cationic Lipid Composition on Gene Silencing Properties of Lipid Nanoparticle Formulations of siRNA in Antigen-Presenting Cells [J].

Basha, Genc ;

Novobrantseva, Tatiana I. ;

Rosin, Nicole ;

Tam, Yuen Yi C. ;

Hafez, Ismail M. ;

Wong, Matthew K. ;

Sugo, Tsukasa ;

Ruda, Vera M. ;

Qin, June ;

Klebanov, Boris ;

Ciufolini, Marco ;

Akinc, Akin ;

Tam, Ying K. ;

Hope, Michael J. ;

Cullis, Pieter R. .

MOLECULAR THERAPY, 2011, 19 (12) :2186-2200

[9] A large-scale chemical modification screen identifies design rules to generate siRNAs with high activity, high stability and low toxicity [J].

Bramsen, Jesper B. ;

Laursen, Maria B. ;

Nielsen, Anne F. ;

Hansen, Thomas B. ;

Bus, Claus ;

Langkjaer, Niels ;

Babu, B. Ravindra ;

Hojland, Torben ;

Abramov, Mikhail ;

Van Aerschot, Arthur ;

Odadzic, Dalibor ;

Smicius, Romualdas ;

Haas, Jens ;

Andree, Cordula ;

Barman, Jharna ;

Wenska, Malgorzata ;

Srivastava, Puneet ;

Zhou, Chuanzheng ;

Honcharenko, Dmytro ;

Hess, Simone ;

Mueller, Elke ;

Bobkov, Georgii V. ;

Mikhailov, Sergey N. ;

Fava, Eugenio ;

Meyer, Thomas F. ;

Chattopadhyaya, Jyoti ;

Zerial, Marino ;

Engels, Joachim W. ;

Herdewijn, Piet ;

Wengel, Jesper ;

Kjems, Jorgen .

NUCLEIC ACIDS RESEARCH, 2009, 37 (09) :2867-2881

[10] Small RNA sorting: matchmaking for Argonautes [J].

Czech, Benjamin ;

Hannon, Gregory J. .

NATURE REVIEWS GENETICS, 2011, 12 (01) :19-31

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