Cell-penetrating peptide-conjugated lipid nanoparticles for siRNA delivery.
Lipid nanoparticles (LNP) modified with cell-penetrating peptides (CPP) were prepared for the delivery of small interfering RNA (siRNA) into cells. Lipid derivatives of CPP derived from protamine were newly synthesized and used to prepare CPP-decorated LNP (CPP-LNP). Encapsulation of siRNA into CPP-LNP improved the stability of the siRNA in serum. Fluorescence-labeled siRNA formulated in CPP-LNP was efficiently internalized into B16F10 murine melanoma cells in a time-dependent manner, although that in LNP without CPP was hardly internalized into these cells. In cells transfected with siRNA in CPP-LNP, most of the siRNA was distributed in the cytoplasm of these cells and did not localize in the lysosomes. Analysis of the endocytotic pathway indicated that CPP-LNP were mainly internalized via macropinocytosis and heparan sulfate-mediated endocytosis. CPP-LNP encapsulating siRNA effectively induced RNA interference-mediated silencing of reporter genes in B16F10 cells expressing luciferase and in HT1080 human fibrosarcoma cells expressing enhanced green fluorescent protein. These data suggest that modification of LNP with the protamine-derived CPP was effective to facilitate internalization of siRNA in the cytoplasm and thereby to enhance gene silencing.
Department of Medical Biochemistry, University of Shizuoka School of Pharmaceutical Sciences, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan. Electronic address: asai@u-shizuoka-ken.ac.jp.
Tomohiro
T
Asai
Department of Medical Biochemistry, University of Shizuoka School of Pharmaceutical Sciences, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
Takuma
T
Tsuzuku
Department of Medical Biochemistry, University of Shizuoka School of Pharmaceutical Sciences, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
Shoya
S
Takahashi
Department of Medical Biochemistry, University of Shizuoka School of Pharmaceutical Sciences, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
Ayaka
A
Okamoto
Department of Life and Materials Engineering, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan.
Takehisa
T
Dewa
Department of Life and Materials Engineering, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan.
Mamoru
M
Nango
Global Formulation Research, Pharmaceutical Science & Technology Core Function Unit, Eisai Product Creation Systems, Eisai Co. Ltd., 5-1-3 Tokodai, Tsukuba 300-2635, Japan.
Kenji
K
Hyodo
Global Formulation Research, Pharmaceutical Science & Technology Core Function Unit, Eisai Product Creation Systems, Eisai Co. Ltd., 5-1-3 Tokodai, Tsukuba 300-2635, Japan.
Hiroshi
H
Ishihara
Global Formulation Research, Pharmaceutical Science & Technology Core Function Unit, Eisai Product Creation Systems, Eisai Co. Ltd., 5-1-3 Tokodai, Tsukuba 300-2635, Japan.
Hiroshi
H
Kikuchi
Department of Medical Biochemistry, University of Shizuoka School of Pharmaceutical Sciences, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
Naoto
N
Oku
D000818:Animals; D045744:Cell Line, Tumor; D057846:Cell-Penetrating Peptides / Q000737:chemistry* / Q000378:metabolism; D004705:Endocytosis; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry* / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry* / Q000378:metabolism; D010873:Pinocytosis; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D057846:Cell-Penetrating Peptides; D010714:Phosphatidylethanolamines; D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
Cell-penetrating peptides; Lipid nanoparticles; Small interfering RNA