title: Insight into mechanisms of cellular uptake of lipid nanoparticles and intracellular release of small RNAs.
abstract: OBJECTIVE
  Understanding mechanisms of cellular uptake and intracellular release would enable better design of nanocarriers for delivery of nucleic acids such as siRNA and microRNA (miRNA).
  
  
  METHODS
  In this study, we investigated cellular pharmacokinetics of siRNA by co-encapsulating fluorescently labeled siRNA and molecular beacon (MB) in four different formulations of cationic lipid nanoparticles (LNPs). A miRNA mimic was also used as a probe for investigating cellular pharmacokinetics, which correlated well with RNAi activities.
  
  
  RESULTS
  We tried to find the best LNP formulation based on the combination of DOTMA and DODMA. When the DOTMA/DODMA ratio was at 5/40, the LNP containing a luciferase siRNA produced the highest gene silencing activity. The superior potency of DOTMA/DODMA could be attributed to higher uptake and improved ability to facilitate siRNA release from endosomes subsequent to uptake.
  
  
  CONCLUSIONS
  Our findings may provide new insights into RNAi transfection pathways and have implications on cationic LNP design.
authors.affiliation: Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Bo
authors.initials: B
authors.lastname: Yu
authors.forename: Xinmei
authors.initials: X
authors.lastname: Wang
authors.forename: Chenguang
authors.initials: C
authors.lastname: Zhou
authors.forename: Lesheng
authors.initials: L
authors.lastname: Teng
authors.forename: Wei
authors.initials: W
authors.lastname: Ren
authors.forename: Zhaogang
authors.initials: Z
authors.lastname: Yang
authors.forename: Chih-Hsin
authors.initials: CH
authors.lastname: Shih
authors.forename: Tianyou
authors.initials: T
authors.lastname: Wang
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Lee
authors.forename: Suoqin
authors.initials: S
authors.lastname: Tang
authors.forename: L James
authors.initials: LJ
authors.lastname: Lee
mesh_terms: D045744:Cell Line, Tumor; D004337:Drug Carriers / Q000737:chemistry*; D004339:Drug Compounding; D065546:Drug Liberation; D004705:Endocytosis / Q000187:drug effects*; D011992:Endosomes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D035683:MicroRNAs / Q000235:genetics; D018613:Microscopy, Confocal; D053758:Nanoparticles / Q000737:chemistry*; D010316:Particle Size; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D034622:RNA Interference / Q000187:drug effects; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000493:pharmacokinetics* / Q000494:pharmacology; D013499:Surface Properties
chemical_list: D004337:Drug Carriers; D008055:Lipids; C531907:MIRN122 microRNA, human; D035683:MicroRNAs; D000644:Quaternary Ammonium Compounds; D034741:RNA, Small Interfering; C473737:dioctyldimethyl ammonium chloride; C054005:N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium
references.citation: Nat Rev Drug Discov. 2010 Oct;9(10):775-89
references.pmid: 20885409
references.citation: Nat Rev Cancer. 2011 Jan;11(1):59-67
references.pmid: 21160526
references.citation: Anticancer Res. 2011 May;31(5):1619-26
references.pmid: 21617218
references.citation: Nucleic Acids Res. 2011 Apr;39(7):e48
references.pmid: 21288880
references.citation: J Control Release. 2013 Dec 28;172(3):690-8
references.pmid: 24121065
references.citation: Nanomedicine. 2013 Feb;9(2):233-46
references.pmid: 22698807
references.citation: J Cell Sci. 2010 Apr 15;123(Pt 8):1183-9
references.pmid: 20356929
references.citation: Nature. 1998 Feb 19;391(6669):806-11
references.pmid: 9486653
references.citation: J Control Release. 2010 Aug 3;145(3):182-95
references.pmid: 20226220
references.citation: J Histochem Cytochem. 2011 Aug;59(8):727-40
references.pmid: 21804077
references.citation: J Biol Chem. 2009 Nov 13;284(46):32015-27
references.pmid: 19726678
references.citation: Nanomedicine. 2013 May;9(4):504-13
references.pmid: 23117046
references.citation: J Control Release. 2010 Aug 3;145(3):289-96
references.pmid: 20417238
references.citation: Mol Pharm. 2011 Oct 3;8(5):1662-8
references.pmid: 21740056
references.citation: Mol Pharm. 2012 Sep 4;9(9):2380-402
references.pmid: 22844998
references.citation: J Control Release. 2012 Aug 10;161(3):757-62
references.pmid: 22641061
references.citation: J Clin Invest. 2012 Aug;122(8):2871-83
references.pmid: 22820288
references.citation: Nanomedicine. 2013 Nov;9(8):1169-80
references.pmid: 23727126
references.citation: Langmuir. 2010 May 4;26(9):6689-94
references.pmid: 20073488
references.citation: Mol Pharm. 2009 Jan-Feb;6(1):221-30
references.pmid: 19183107
references.citation: AAPS J. 2009 Mar;11(1):195-203
references.pmid: 19296227
references.citation: Biochemistry. 2006 Sep 5;45(35):10614-23
references.pmid: 16939213
references.citation: ACS Nano. 2012 Jul 24;6(7):6133-41
references.pmid: 22693946
references.citation: Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):1864-9
references.pmid: 20080679
references.citation: Nat Biotechnol. 2010 Feb;28(2):172-6
references.pmid: 20081866
references.citation: Mol Ther. 2011 Sep;19(9):1688-94
references.pmid: 21750531
references.citation: J Med Chem. 2010 Nov 25;53(22):7887-901
references.pmid: 20806941
references.citation: Nat Rev Genet. 2007 Mar;8(3):173-84
references.pmid: 17304245
references.citation: Biomaterials. 2012 Sep;33(25):5924-34
references.pmid: 22652024
references.citation: Hepatology. 2014 Feb;59(2):555-66
references.pmid: 24038073
references.citation: Nucleic Acids Res. 2007;35(18):e124
references.pmid: 17890733
references.citation: Mol Ther. 2008 Feb;16(2):324-32
references.pmid: 18180773
references.citation: Biol Chem. 2012 Jan;393(1-2):23-35
references.pmid: 22628296
references.citation: Mol Biosyst. 2011 Jul;7(7):2110-3
references.pmid: 21541377
references.citation: Mol Ther. 2010 Jan;18(1):171-80
references.pmid: 19738601
references.citation: Biomaterials. 2012 Nov;33(31):7834-48
references.pmid: 22874502