Pre-administration of PepFect6-microRNA-146a nanocomplexes inhibits inflammatory responses in keratinocytes and in a mouse model of irritant contact dermatitis.
The skin is a difficult to access tissue for efficient delivery of large and/or charged macromolecules, including therapeutic DNA and RNA oligonucleotides. Cell-penetrating peptide PepFect6 (PF6) has been shown to be suitable transport vehicle for siRNAs in cell culture and systemically in vivo in mice. MiR-146a is known as anti-inflammatory miRNA that inhibits multiple factors from the nuclear factor (NF)-κB pathway in various cell types, including keratinocytes. In this study, PF6 was shown to form unimodal nanocomplexes with miR-146a mimic that entered into human primary keratinocytes, where miR-146a inhibited the expression of its direct targets from the NF-κB pathway and the genes known to be activated by NF-κB, C-C motif ligand (CCL)5 and interleukin (IL)-8. The transfection of miR-146a mimic with PF6 was more efficient in sub-confluent keratinocyte cultures, affected keratinocyte proliferation less and had similar effect on cell viability when compared with a lipid based agent. Subcutaneous pre-administration of PF6-miR-146a nanocomplexes attenuated ear-swelling and reduced the expression of pro-inflammatory cytokines and chemokines IL-6, CCL11, CCL24 and C-X-C motif ligand 1 (CXCL1) in a mouse model of irritant contact dermatitis. Our data demonstrates that PF6-miR-146a nanoparticles might have potential in the development of therapeutics to target inflammatory skin diseases.
Institute of Biomedicine and Translational Medicine, University of Tartu, Estonia; Institute of Molecular and Cell Biology, University of Tartu, Estonia.
Egon
E
Urgard
Institute of Molecular and Cell Biology, University of Tartu, Estonia.
Annely
A
Lorents
Institute of Molecular and Cell Biology, University of Tartu, Estonia.
Mariliis
M
Klaas
Institute of Molecular and Cell Biology, University of Tartu, Estonia.
Kärt
K
Padari
Institute of Molecular and Cell Biology, University of Tartu, Estonia.
Janeli
J
Viil
Institute of Biomedicine and Translational Medicine, University of Tartu, Estonia; Institute of Molecular and Cell Biology, University of Tartu, Estonia.
Toomas
T
Runnel
Institute of Technology, University of Tartu, Estonia.
Kent
K
Langel
Department of Dermatology and Venereology, University of Tartu, Tartu, Estonia; Dermatology Clinic, Tartu University Hospital, Tartu, Estonia.
Külli
K
Kingo
Institute of Biomedicine and Translational Medicine, University of Tartu, Estonia; Department of Medicine, Vanderbilt University Medical Center, United States.
Eric
E
Tkaczyk
Institute of Technology, University of Tartu, Estonia; Department of Neurochemistry, Stockholm University, Sweden.
Ülo
Ü
Langel
Institute of Molecular and Cell Biology, University of Tartu, Estonia.
Toivo
T
Maimets
Institute of Molecular and Cell Biology, University of Tartu, Estonia; Department of Bioscience, Karolinska Institute, Sweden.
Viljar
V
Jaks
Institute of Molecular and Cell Biology, University of Tartu, Estonia.
Margus
M
Pooga
Institute of Biomedicine and Translational Medicine, University of Tartu, Estonia. Electronic address: ana.rebane@ut.ee.
Ana
A
Rebane
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage* / Q000737:chemistry / Q000627:therapeutic use; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology; D003877:Dermatitis, Contact / Q000188:drug therapy / Q000276:immunology; D004195:Disease Models, Animal; D006801:Humans; D007509:Irritants; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism; D055666:Lipopeptides / Q000008:administration & dosage* / Q000737:chemistry / Q000627:therapeutic use; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics / Q000627:therapeutic use; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry / Q000627:therapeutic use / Q000648:ultrastructure; D011804:Quinolines / Q000008:administration & dosage* / Q000737:chemistry / Q000627:therapeutic use; D013755:Tetradecanoylphorbol Acetate
D000893:Anti-Inflammatory Agents; D016207:Cytokines; D007509:Irritants; D055666:Lipopeptides; D035683:MicroRNAs; C534190:Mirn146 microRNA, mouse; C559845:PepFect6 peptide; D011804:Quinolines; D013755:Tetradecanoylphorbol Acetate
Allergy; MicroRNA; Non-coding RNA; SiRNA