Design of a peptide-based vector, PepFect6, for efficient delivery of siRNA in cell culture and systemically in vivo.
While small interfering RNAs (siRNAs) have been rapidly appreciated to silence genes, efficient and non-toxic vectors for primary cells and for systemic in vivo delivery are lacking. Several siRNA-delivery vehicles, including cell-penetrating peptides (CPPs), have been developed but their utility is often restricted by entrapment following endocytosis. Hence, developing CPPs that promote endosomal escape is a prerequisite for successful siRNA implementation. We here present a novel CPP, PepFect 6 (PF6), comprising the previously reported stearyl-TP10 peptide, having pH titratable trifluoromethylquinoline moieties covalently incorporated to facilitate endosomal release. Stable PF6/siRNA nanoparticles enter entire cell populations and rapidly promote endosomal escape, resulting in robust RNAi responses in various cell types (including primary cells), with minimal associated transcriptomic or proteomic changes. Furthermore, PF6-mediated delivery is independent of cell confluence and, in most cases, not significantly hampered by serum proteins. Finally, these nanoparticles promote strong RNAi responses in different organs following systemic delivery in mice without any associated toxicity. Strikingly, similar knockdown in liver is achieved by PF6/siRNA nanoparticles and siRNA injected by hydrodynamic infusion, a golden standard technique for liver transfection. These results imply that the peptide, in addition to having utility for RNAi screens in vitro, displays therapeutic potential.
Department of Neurochemistry, Stockholm University, 106 92 Stockholm, Sweden. samir.el-andaloussi@ki.se
Samir E L
SE
Andaloussi
Taavi
T
Lehto
Imre
I
Mäger
Katri
K
Rosenthal-Aizman
Iulian I
II
Oprea
Oscar E
OE
Simonson
Helena
H
Sork
Kariem
K
Ezzat
Dana M
DM
Copolovici
Kaido
K
Kurrikoff
Joana R
JR
Viola
Eman M
EM
Zaghloul
Rannar
R
Sillard
Henrik J
HJ
Johansson
Fatouma
F
Said Hassane
Peter
P
Guterstam
Julia
J
Suhorutšenko
Pedro M D
PM
Moreno
Nikita
N
Oskolkov
Jonas
J
Hälldin
Ulf
U
Tedebark
Andres
A
Metspalu
Bernard
B
Lebleu
Janne
J
Lehtiö
C I Edvard
CI
Smith
Ulo
U
Langel
D000818:Animals; D057846:Cell-Penetrating Peptides / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D002478:Cells, Cultured; D011992:Endosomes / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents; D018836:Inflammation Mediators / Q000378:metabolism; D008055:Lipids; D055666:Lipopeptides / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity; D011804:Quinolines / Q000737:chemistry* / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage*
D057846:Cell-Penetrating Peptides; D007202:Indicators and Reagents; D018836:Inflammation Mediators; D008055:Lipids; C086724:Lipofectamine; D055666:Lipopeptides; C559845:PepFect6 peptide; D011804:Quinolines; D034741:RNA, Small Interfering
Adv Drug Deliv Rev. 2008 Mar 1;60(4-5):530-6
18155315
Cancer Biol Ther. 2009 Dec;8(23):2306-13
19901546
Nature. 2009 Jan 22;457(7228):426-33
19158789
Nat Biotechnol. 2008 May;26(5):561-9
18438401
Proteomics. 2008 Aug;8(15):3008-18
18654985
Nat Rev Drug Discov. 2009 Feb;8(2):129-38
19180106
Cell. 2008 Aug 22;134(4):577-86
18691745
J Control Release. 2009 Oct 15;139(2):127-32
19540888
Nucleic Acids Res. 2009 Aug;37(14):4559-69
19483097
EMBO Mol Med. 2009 Jun;1(3):142-51
20049714
Nature. 2007 Jul 5;448(7149):39-43
17572664
Mol Ther. 2006 Mar;13(3):494-505
16343994
Curr Pharm Des. 2005;11(28):3597-611
16305497
Nat Biotechnol. 1998 Sep;16(9):857-61
9743120
Nat Biotechnol. 2009 Jun;27(6):567-71
19448630
J Control Release. 2010 May 10;143(3):311-7
20080139
Mol Ther. 2007 Sep;15(9):1663-9
17579574
Nature. 2008 Apr 3;452(7187):591-7
18368052
FASEB J. 2007 Sep;21(11):2664-71
17463227
Cell. 2005 Sep 23;122(6):947-56
16153702
Nature. 2008 Oct 30;455(7217):1251-4
18820680
Nature. 2002 Jul 4;418(6893):38-9
12097900
Bioconjug Chem. 2008 Dec;19(12):2535-42
19012426
Nature. 2006 May 4;441(7089):111-4
16565705
Gene Ther. 1999 Jul;6(7):1258-66
10455434
J Control Release. 2009 Mar 19;134(3):221-7
19105971
Toxicology. 2009 Nov 30;265(3):87-95
19799958
Hum Gene Ther. 2008 Feb;19(2):111-24
18230025
J Biol Chem. 1997 Oct 24;272(43):26833-40
9341114
Nat Med. 2005 Mar;11(3):263-70
15723075
Biochem J. 2007 Oct 15;407(2):285-92
17627607
Nature. 1998 Feb 19;391(6669):806-11
9486653
Nat Rev Drug Discov. 2007 Jun;6(6):443-53
17541417
Nucleic Acids Res. 1999 Sep 1;27(17):3510-7
10446241
Oligonucleotides. 2008 Jun;18(2):187-200
18637735
Nat Rev Drug Discov. 2010 Aug;9(8):615-27
20616808
Adv Drug Deliv Rev. 2008 Mar 1;60(4-5):537-47
18037526
Biochem Soc Trans. 2007 Aug;35(Pt 4):807-10
17635153
J Control Release. 2004 May 31;97(1):133-41
15147811
Pharm Res. 2002 May;19(5):676-80
12069172
Biomaterials. 2010 Feb;31(5):998-1006
19850333
Mol Ther. 2010 Sep;18(9):1657-66
20628357
J Med Chem. 2006 Nov 2;49(22):6522-31
17064070
Br J Pharmacol. 2009 May;157(2):195-206
19309362
Expert Opin Drug Deliv. 2009 Nov;6(11):1195-205
19831582