title: Receptor-targeted liposome-peptide-siRNA nanoparticles represent an efficient delivery system for MRTF silencing in conjunctival fibrosis.
abstract: There is increasing evidence that the Myocardin-related transcription factor/Serum response factor (MRTF/SRF) pathway plays a key role in fibroblast activation and that knocking down MRTF can lead to reduced scarring and fibrosis. Here, we have developed a receptor-targeted liposome-peptide-siRNA nanoparticle as a non-viral delivery system for MRTF-B siRNA in conjunctival fibrosis. Using 50 nM siRNA, the MRTF-B gene was efficiently silenced by 76% and 72% with LYR and LER nanoparticles, respectively. The silencing efficiency was low when non-targeting peptides or siRNA alone or liposome-siRNA alone were used. LYR and LER nanoparticles also showed higher silencing efficiency than PEGylated LYR-P and LER-P nanoparticles. The nanoparticles were not cytotoxic using different liposomes, targeting peptides, and 50 nM siRNA. Three-dimensional fibroblast-populated collagen matrices were also used as a functional assay to measure contraction in vitro, and showed that MRTF-B LYR nanoparticles completely blocked matrix contraction after a single transfection treatment. In conclusion, this is the first study to develop and show that receptor-targeted liposome-peptide-siRNA nanoparticles represent an efficient and safe non-viral siRNA delivery system that could be used to prevent fibrosis after glaucoma filtration surgery and other contractile scarring conditions in the eye.
authors.affiliation: National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Yu-Wai-Man
authors.affiliation: Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, London, United Kingdom.
authors.forename: Aristides D
authors.initials: AD
authors.lastname: Tagalakis
authors.affiliation: Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, London, United Kingdom.
authors.forename: Maria D
authors.initials: MD
authors.lastname: Manunta
authors.affiliation: Wolfson Centre for Gene Therapy of Childhood Disease, UCL Institute of Child Health, London, United Kingdom.
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Hart
authors.affiliation: National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom.
authors.forename: Peng T
authors.initials: PT
authors.lastname: Khaw
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D004337:Drug Carriers / Q000737:chemistry*; D005128:Eye Diseases / Q000378:metabolism / Q000473:pathology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005355:Fibrosis; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000737:chemistry / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D004337:Drug Carriers; D008081:Liposomes; D010455:Peptides; D034741:RNA, Small Interfering; D014157:Transcription Factors; D011092:Polyethylene Glycols