title: Visualization of plasmid delivery to keratinocytes in mouse and human epidermis.
abstract: The accessibility of skin makes it an ideal target organ for nucleic acid-based therapeutics; however, effective patient-friendly delivery remains a major obstacle to clinical utility. A variety of limited and inefficient methods of delivering nucleic acids to keratinocytes have been demonstrated; further advances will require well-characterized reagents, rapid noninvasive assays of delivery, and well-developed skin model systems. Using intravital fluorescence and bioluminescence imaging and a standard set of reporter plasmids we demonstrate transfection of cells in mouse and human xenograft skin using intradermal injection and two microneedle array delivery systems. Reporter gene expression could be detected in individual keratinocytes, in real-time, in both mouse skin as well as human skin xenografts. These studies revealed that non-invasive intravital imaging can be used as a guide for developing gene delivery tools, establishing a benchmark for comparative testing of nucleic acid skin delivery technologies.
authors.affiliation: Molecular Imaging Program at Stanford (MIPS), Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Emilio
authors.initials: E
authors.lastname: González-González
authors.forename: Yeu-Chun
authors.initials: YC
authors.lastname: Kim
authors.forename: Tycho J
authors.initials: TJ
authors.lastname: Speaker
authors.forename: Robyn P
authors.initials: RP
authors.lastname: Hickerson
authors.forename: Ryan
authors.initials: R
authors.lastname: Spitler
authors.forename: James C
authors.initials: JC
authors.lastname: Birchall
authors.forename: Maria Fernanda
authors.initials: MF
authors.lastname: Lara
authors.forename: Rong-Hua
authors.initials: RH
authors.lastname: Hu
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Liang
authors.forename: Nancy
authors.initials: N
authors.lastname: Kirkiles-Smith
authors.forename: Mark R
authors.initials: MR
authors.lastname: Prausnitz
authors.forename: Leonard M
authors.initials: LM
authors.lastname: Milstone
authors.forename: Christopher H
authors.initials: CH
authors.lastname: Contag
authors.forename: Roger L
authors.initials: RL
authors.lastname: Kaspar
mesh_terms: D000818:Animals; D016503:Drug Delivery Systems; D004817:Epidermis / Q000378:metabolism; D005260:Female; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements; D051379:Mice; D008845:Microinjections; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016038:Skin Transplantation; D014162:Transfection; D014183:Transplantation, Heterologous
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D008156:Luciferases