title: Inhibition of CD44 gene expression in human skin models, using self-delivery short interfering RNA administered by dissolvable microneedle arrays.
abstract: Treatment of skin disorders with short interfering RNA (siRNA)-based therapeutics requires the development of effective delivery methodologies that reach target cells in affected tissues. Successful delivery of functional siRNA to the epidermis requires (1) crossing the stratum corneum, (2) transfer across the keratinocyte membrane, followed by (3) incorporation into the RNA-induced silencing complex. We have previously demonstrated that treatment with microneedle arrays loaded with self-delivery siRNA (sd-siRNA) can achieve inhibition of reporter gene expression in a transgenic mouse model. Furthermore, treatment of human cultured epidermal equivalents with sd-siRNA resulted in inhibition of target gene expression. Here, we demonstrate inhibition of CD44, a gene that is uniformly expressed throughout the epidermis, by sd-siRNA both in vitro (cultured human epidermal skin equivalents) and in vivo (full-thickness human skin equivalents xenografted on immunocompromised mice). Treatment of human skin equivalents with CD44 sd-siRNA markedly decreased CD44 mRNA levels, which led to a reduction of the target protein as confirmed by immunodetection in epidermal equivalent sections with a CD44-specific antibody. Taken together, these results demonstrate that sd-siRNA, delivered by microneedle arrays, can reduce expression of a targeted endogenous gene in a human skin xenograft model.
authors.affiliation: TransDerm, Santa Cruz, CA 95060, USA.
authors.forename: Maria Fernanda
authors.initials: MF
authors.lastname: Lara
authors.forename: Emilio
authors.initials: E
authors.lastname: González-González
authors.forename: Tycho J
authors.initials: TJ
authors.lastname: Speaker
authors.forename: Robyn P
authors.initials: RP
authors.lastname: Hickerson
authors.forename: Devin
authors.initials: D
authors.lastname: Leake
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; D002478:Cells, Cultured; D005260:Female; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D009339:Needles; D019904:Polymethyl Methacrylate; D011142:Polyvinyl Alcohol; D034741:RNA, Small Interfering / Q000008:administration & dosage*; D012867:Skin / Q000378:metabolism*; D012995:Solubility; D014183:Transplantation, Heterologous
chemical_list: C497483:CD44 protein, human; D018960:Hyaluronan Receptors; D034741:RNA, Small Interfering; D011142:Polyvinyl Alcohol; D019904:Polymethyl Methacrylate