title: Self-assembled Micelle Interfering RNA for Effective and Safe Targeting of Dysregulated Genes in Pulmonary Fibrosis.
abstract: The siRNA silencing approach has long been used as a method to regulate the expression of specific target genes in vitro and in vivo. However, the effectiveness of delivery and the nonspecific immune-stimulatory function of siRNA are the limiting factors for therapeutic applications of siRNAs. To overcome these limitations, we developed self-assembled micelle inhibitory RNA (SAMiRNA) nanoparticles made of individually biconjugated siRNAs with a hydrophilic polymer and lipid on their ends and characterized their stability, immune-stimulatory function, and in vivo silencing efficacy. SAMiRNAs form very stable nanoparticles with no significant degradation in size distribution and polydispersity index over 1 year. Overnight incubation of SAMiRNAs (3 μm) on murine peripheral blood mononuclear cells did not cause any significant elaboration of innate immune cytokines such as TNF-α, IL-12, or IL-6, whereas unmodified siRNAs or liposomes or liposome complexes significantly stimulated the expression of these cytokines. Last, the in vivo silencing efficacy of SAMiRNAs was evaluated by targeting amphiregulin and connective tissue growth factor in bleomycin or TGF-β transgenic animal models of pulmonary fibrosis. Intratracheal or intravenous delivery two or three times of amphiregulin or connective tissue growth factor SAMiRNAs significantly reduced the bleomycin- or TGF-β-stimulated collagen accumulation in the lung and substantially restored the lung function of TGF-β transgenic mice. This study demonstrates that SAMiRNA nanoparticle is a less toxic, stable siRNA silencing platform for efficient in vivo targeting of genes implicated in the pathogenesis of pulmonary fibrosis.
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Pyoung Oh
authors.initials: PO
authors.lastname: Yoon
authors.affiliation: the Department of Molecular Microbiology and Immunology.
authors.forename: Jin Wook
authors.initials: JW
authors.lastname: Park
authors.affiliation: the Department of Molecular Microbiology and Immunology.
authors.forename: Chang-Min
authors.initials: CM
authors.lastname: Lee
authors.affiliation: the Inhalation Toxicology Center, Korea Institute of Toxicology, Jeongeup Campus, Jeollabuk-do 580-185, Korea, and.
authors.forename: Sung Hwan
authors.initials: SH
authors.lastname: Kim
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Han-Na
authors.initials: HN
authors.lastname: Kim
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Youngho
authors.initials: Y
authors.lastname: Ko
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Seon Joo
authors.initials: SJ
authors.lastname: Bae
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Sungil
authors.initials: S
authors.lastname: Yun
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Jun Hong
authors.initials: JH
authors.lastname: Park
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Taewoo
authors.initials: T
authors.lastname: Kwon
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Woo Seok
authors.initials: WS
authors.lastname: Kim
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Jiyoung
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Medicine, Alpert Medical School, Brown University, Providence, Rhode Island 02912.
authors.forename: Qing
authors.initials: Q
authors.lastname: Lu
authors.affiliation: the Division of Allergy and Clinical Immunology, Department of Internal Medicine, Seoul National University Hospital, Seoul 110-744, Korea.
authors.forename: Hye-Ryun
authors.initials: HR
authors.lastname: Kang
authors.affiliation: Department of Medicine, Alpert Medical School, Brown University, Providence, Rhode Island 02912.
authors.forename: Won-Kyung
authors.initials: WK
authors.lastname: Cho
authors.affiliation: the Department of Molecular Microbiology and Immunology.
authors.forename: Jack A
authors.initials: JA
authors.lastname: Elias
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Joo-Sung
authors.initials: JS
authors.lastname: Yang
authors.affiliation: From the Bioneer Corp., Daedeok-gu, Daejeon 306-220, Korea.
authors.forename: Han-Oh
authors.initials: HO
authors.lastname: Park
authors.affiliation: the Inhalation Toxicology Center, Korea Institute of Toxicology, Jeongeup Campus, Jeollabuk-do 580-185, Korea, and khlee@kitox.re.kr.
authors.forename: Kyuhong
authors.initials: K
authors.lastname: Lee
authors.affiliation: the Department of Molecular Microbiology and Immunology, chun_lee@brown.edu.
authors.forename: Chun Geun
authors.initials: CG
authors.lastname: Lee
mesh_terms: D066258:Amphiregulin; D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D055513:Connective Tissue Growth Factor / Q000235:genetics / Q000378:metabolism; D066255:EGF Family of Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D055785:Gene Knockdown Techniques / Q000379:methods; D015316:Genetic Therapy; D008168:Lung / Q000378:metabolism / Q000473:pathology; D008297:Male; D008810:Mice, Inbred C57BL; D008823:Micelles; D053758:Nanoparticles; D011658:Pulmonary Fibrosis / Q000235:genetics / Q000628:therapy*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics* / Q000493:pharmacokinetics; D014018:Tissue Distribution
chemical_list: D066258:Amphiregulin; C587849:Areg protein, mouse; C526318:CCN2 protein, mouse; D066255:EGF Family of Proteins; D008823:Micelles; D034741:RNA, Small Interfering; D055513:Connective Tissue Growth Factor; D003094:Collagen
keywords: amphiregulin; connective tissue growth factor (CTGF); cytokine; fibrosis; pulmonary fibrosis; siRNA
references.citation: Expert Rev Clin Pharmacol. 2008 Jul;1(4):547-58
references.pmid: 24410556
references.citation: J Mol Med (Berl). 2005 Oct;83(10):764-73
references.pmid: 16028076
references.citation: J Biol Chem. 2012 Dec 7;287(50):41991-2000
references.pmid: 23086930
references.citation: J Clin Invest. 1997 Aug 15;100(4):768-76
references.pmid: 9259574
references.citation: N Engl J Med. 2014 May 29;370(22):2083-92
references.pmid: 24836312
references.citation: Biophys J. 2001 May;80(5):2310-26
references.pmid: 11325732
references.citation: Ann Intern Med. 2001 Jan 16;134(2):136-51
references.pmid: 11177318
references.citation: Am J Respir Crit Care Med. 2010 Oct 1;182(7):918-28
references.pmid: 20558631
references.citation: Curr Pharm Des. 2003;9(1):39-49
references.pmid: 12570673
references.citation: Lab Anim. 2010 Jan;44(1):20-4
references.pmid: 19880440
references.citation: Toxicol Res. 2010 Sep;26(3):217-22
references.pmid: 24278527
references.citation: Arthritis Rheum. 2009 Jul;60(7):2142-55
references.pmid: 19565505
references.citation: J Med Chem. 2014 Feb 27;57(4):1138-46
references.pmid: 24047101
references.citation: Oligonucleotides. 2009 Jun;19(2):89-102
references.pmid: 19441890
references.citation: RNA. 2004 May;10(5):766-71
references.pmid: 15100431
references.citation: Chest. 2002 Dec;122(6 Suppl):289S-293S
references.pmid: 12475802
references.citation: Nat Nanotechnol. 2012 Jun;7(6):389-93
references.pmid: 22659608
references.citation: Fibrogenesis Tissue Repair. 2011 Feb 01;4(1):4
references.pmid: 21284856
references.citation: Biochemistry. 1992 Sep 22;31(37):9025-30
references.pmid: 1390689
references.citation: N Engl J Med. 2014 May 29;370(22):2071-82
references.pmid: 24836310
references.citation: Fibrogenesis Tissue Repair. 2012 Jun 06;5(Suppl 1):S24
references.pmid: 23259531
references.citation: Bioconjug Chem. 2010 May 19;21(5):797-802
references.pmid: 20397686
references.citation: J Control Release. 2003 Dec 5;93(2):183-91
references.pmid: 14636724
references.citation: Am J Respir Cell Mol Biol. 1996 Feb;14(2):131-8
references.pmid: 8630262
references.citation: J Mol Cell Biol. 2013 Oct;5(5):323-35
references.pmid: 23929714
references.citation: Nat Rev Drug Discov. 2009 Feb;8(2):129-38
references.pmid: 19180106
references.citation: Mol Ther. 2011 Feb;19(2):381-5
references.pmid: 21119623
references.citation: J Exp Med. 2004 Aug 2;200(3):377-89
references.pmid: 15289506
references.citation: Am J Physiol Lung Cell Mol Physiol. 2003 Sep;285(3):L527-39
references.pmid: 12598227
references.citation: Bioconjug Chem. 2009 Jan;20(1):5-14
references.pmid: 19053311
references.citation: N Engl J Med. 2014 Aug 21;371(8):783-4
references.pmid: 25147884
references.citation: Nat Rev Drug Discov. 2002 Jul;1(7):503-14
references.pmid: 12120257
references.citation: Nat Mater. 2013 Nov;12(11):967-77
references.pmid: 24150415
references.citation: Proc Natl Acad Sci U S A. 2012 Apr 3;109(14):E797-803
references.pmid: 22421433
references.citation: J Exp Med. 2009 May 11;206(5):1149-66
references.pmid: 19414556
references.citation: Proc Am Thorac Soc. 2012 Jul;9(3):111-6
references.pmid: 22802283
references.citation: Biomaterials. 2012 Oct;33(29):7138-50
references.pmid: 22796160
references.citation: J Exp Med. 2007 May 14;204(5):1083-93
references.pmid: 17485510
references.citation: Mol Ther. 2006 Apr;13(4):644-70
references.pmid: 16481219
references.citation: J Drug Target. 2012 Nov;20(9):724-35
references.pmid: 22994300
references.citation: Thorax. 2001 Dec;56(12):907-15
references.pmid: 11713352
references.citation: Drug Discov Today. 2008 Oct;13(19-20):842-55
references.pmid: 18614389
references.citation: J Clin Invest. 1999 Jul;104(1):5-11
references.pmid: 10393693
references.citation: In Vitro Cell Dev Biol Anim. 1995 Oct;31(9):684-91
references.pmid: 8564079
references.citation: Eur J Pharm Biopharm. 2011 Apr;77(3):438-49
references.pmid: 21093588
references.citation: Oligonucleotides. 2008 Sep;18(3):201-12
references.pmid: 18729822
references.citation: Am J Respir Cell Mol Biol. 2000 Aug;23(2):204-12
references.pmid: 10919987