title: In vivo therapeutic potential of Dicer-hunting siRNAs targeting infectious hepatitis C virus.
abstract: The development of RNA interference (RNAi)-based therapy faces two major obstacles: selecting small interfering RNA (siRNA) sequences with strong activity, and identifying a carrier that allows efficient delivery to target organs. Additionally, conservative region at nucleotide level must be targeted for RNAi in applying to virus because hepatitis C virus (HCV) could escape from therapeutic pressure with genome mutations. In vitro preparation of Dicer-generated siRNAs targeting a conserved, highly ordered HCV 5' untranslated region are capable of inducing strong RNAi activity. By dissecting the 5'-end of an RNAi-mediated cleavage site in the HCV genome, we identified potent siRNA sequences, which we designate as Dicer-hunting siRNAs (dh-siRNAs). Furthermore, formulation of the dh-siRNAs in an optimized multifunctional envelope-type nano device inhibited ongoing infectious HCV replication in human hepatocytes in vivo. Our efforts using both identification of optimal siRNA sequences and delivery to human hepatocytes suggest therapeutic potential of siRNA for a virus.
authors.affiliation: 1] Department of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, Japan [2] Division of Gastroenterology, Showa University Fujigaoka Hospital, Yokohama, Japan [3] Present address, Department of Virology & Liver Unit, Nagoya City University Graduate School of Medical Sciences, Kawasumi, Mizuho, Nagoya 467-8601, Japan [4].
authors.forename: Tsunamasa
authors.initials: T
authors.lastname: Watanabe
authors.affiliation: 1] Laboratory of Innovative Nanomedicine, Faculty of Pharmaceutical Sciences, Hokkaido University, Hokkaido 060-0812, Japan [2].
authors.forename: Hiroto
authors.initials: H
authors.lastname: Hatakeyama
authors.affiliation: 1] Department of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, Japan [2].
authors.forename: Chiho
authors.initials: C
authors.lastname: Matsuda-Yasui
authors.affiliation: 1] Laboratory of Innovative Nanomedicine, Faculty of Pharmaceutical Sciences, Hokkaido University, Hokkaido 060-0812, Japan [2].
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Sato
authors.affiliation: Kamakura Research Laboratories, Chugai Pharmaceutical Co., Ltd., Kamakura, Kanagawa 247-8530, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Sudoh
authors.affiliation: Department of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, Japan.
authors.forename: Asako
authors.initials: A
authors.lastname: Takagi
authors.affiliation: 1] Department of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, Japan [2] Division of Gastroenterology, Showa University Fujigaoka Hospital, Yokohama, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Hirata
authors.affiliation: Department of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Ohtsuki
authors.affiliation: Advanced Medical Research Laboratory, Mitsubishi Tanabe Pharma Corporation, 1000, Kamoshida-cho, Aoba-ku, Yokohama 227-0033, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Arai
authors.affiliation: Division of Gastroenterology, Showa University Fujigaoka Hospital, Yokohama, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Inoue
authors.affiliation: Laboratory of Innovative Nanomedicine, Faculty of Pharmaceutical Sciences, Hokkaido University, Hokkaido 060-0812, Japan.
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Harashima
authors.affiliation: Department of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo 156-8506, Japan.
authors.forename: Michinori
authors.initials: M
authors.lastname: Kohara
mesh_terms: D020121:5' Untranslated Regions; D000818:Animals; D002460:Cell Line; D004195:Disease Models, Animal; D020868:Gene Silencing; D016679:Genome, Viral; D016174:Hepacivirus / Q000235:genetics*; D006526:Hepatitis C / Q000378:metabolism* / Q000628:therapy / Q000821:virology*; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D012367:RNA, Viral; D043244:Ribonuclease III / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D020121:5' Untranslated Regions; D034741:RNA, Small Interfering; D012367:RNA, Viral; D043244:Ribonuclease III
references.citation: Nucleic Acids Res. 2005 Feb 01;33(2):796-804
references.pmid: 15687388
references.citation: Nat Struct Mol Biol. 2005 May;12(5):469-70
references.pmid: 15852021
references.citation: Nat Biotechnol. 2008 May;26(5):561-9
references.pmid: 18438401
references.citation: Biotechnol Bioeng. 2008 Jul 1;100(4):744-55
references.pmid: 18306428
references.citation: J Mol Biol. 2005 May 13;348(4):871-81
references.pmid: 15843019
references.citation: Am J Pathol. 2010 Jan;176(1):14-21
references.pmid: 19948827
references.citation: J Virol. 1992 Mar;66(3):1476-83
references.pmid: 1310759
references.citation: Nat Biotechnol. 2005 Feb;23(2):222-6
references.pmid: 15619617
references.citation: J Control Release. 2014 Jan 10;173:43-50
references.pmid: 24511611
references.citation: Nucleic Acids Res. 2013 Dec;41(22):10659-67
references.pmid: 24030710
references.citation: Mol Ther. 2010 Jul;18(7):1357-64
references.pmid: 20461061
references.citation: Nat Rev Mol Cell Biol. 2008 Jan;9(1):22-32
references.pmid: 18073770
references.citation: Nat Biotechnol. 2004 Mar;22(3):326-30
references.pmid: 14758366
references.citation: Gene Ther. 2006 Jun;13(11):883-92
references.pmid: 16496015
references.citation: Angew Chem Int Ed Engl. 2012 Aug 20;51(34):8529-33
references.pmid: 22782619
references.citation: Nature. 2001 May 24;411(6836):494-8
references.pmid: 11373684
references.citation: Arch Virol. 2011 Feb;156(2):295-304
references.pmid: 21246385
references.citation: Proc Natl Acad Sci U S A. 1984 Mar;81(6):1715-8
references.pmid: 6584905
references.citation: Nature. 2010 May 6;465(7294):96-100
references.pmid: 20410884
references.citation: J Viral Hepat. 2006 Nov;13(11):756-61
references.pmid: 17052275
references.citation: Cell. 2007 Jul 13;130(1):101-12
references.pmid: 17632058
references.citation: Adv Drug Deliv Rev. 2008 Mar 1;60(4-5):559-71
references.pmid: 18037529
references.citation: J Virol. 2004 Mar;78(5):2601-5
references.pmid: 14963165
references.citation: PLoS One. 2012;7(12):e51656
references.pmid: 23284733
references.citation: FEBS Lett. 1985 Jan 1;179(1):148-54
references.pmid: 2578112
references.citation: Nat Biotechnol. 2005 Feb;23(2):227-31
references.pmid: 15619616
references.citation: Nat Med. 2001 Aug;7(8):927-33
references.pmid: 11479625
references.citation: Nat Biotechnol. 2010 Feb;28(2):172-6
references.pmid: 20081866
references.citation: J Virol. 2005 Jun;79(11):7050-8
references.pmid: 15890944
references.citation: Nucleic Acids Res. 2004 Feb 09;32(3):936-48
references.pmid: 14769950
references.citation: Nat Chem Biol. 2005 Nov;1(6):333-7
references.pmid: 16408072
references.citation: Nature. 2002 Jul 11;418(6894):244-51
references.pmid: 12110901
references.citation: Nat Biotechnol. 2008 May;26(5):578-83
references.pmid: 18438400
references.citation: Nat Rev Drug Discov. 2007 Jun;6(6):443-53
references.pmid: 17541417
references.citation: Nature. 2005 Aug 18;436(7053):953-60
references.pmid: 16107835
references.citation: J Biol Chem. 2004 Apr 9;279(15):14531-41
references.pmid: 14747471
references.citation: Mol Ther. 2013 Jun;21(6):1195-203
references.pmid: 23568259
references.citation: Nat Biotechnol. 2005 Nov;23(11):1440-4
references.pmid: 16258545
references.citation: Nat Med. 2005 Jul;11(7):791-6
references.pmid: 15951748
references.citation: N Engl J Med. 2006 Dec 7;355(23):2444-51
references.pmid: 17151366
references.citation: Nature. 2009 Jan 22;457(7228):405-12
references.pmid: 19158786
references.citation: Nat Methods. 2006 Sep;3(9):670-6
references.pmid: 16929310
references.citation: Biochem Biophys Res Commun. 2005 Mar 4;328(1):57-62
references.pmid: 15670750
references.citation: Proc Natl Acad Sci U S A. 2008 Jan 15;105(2):512-7
references.pmid: 18178619
references.citation: J Virol. 1999 Feb;73(2):1165-74
references.pmid: 9882318
references.citation: Nucleic Acids Res. 2006 Jul 1;34(Web Server issue):W448-50
references.pmid: 16845046
references.citation: J Control Release. 2012 Nov 10;163(3):267-76
references.pmid: 23000694
references.citation: J Med Virol. 2013 Feb;85(2):241-9
references.pmid: 23192857
references.citation: BMC Bioinformatics. 2009 Nov 30;10:392
references.pmid: 19948054
references.citation: Hepatology. 2007 Apr;45(4):921-8
references.pmid: 17393519