In vivo gene silencing following non-invasive siRNA delivery into the skin using a novel topical formulation

Vikas Hegde, Robyn P. Hickerson, Sitheswaran Nainamalai, Paul A. Campbell, Frances J. D. Smith, W. H. Irwin McLean (Lead / Corresponding author), Deena M. Leslie Pedrioli (Lead / Corresponding author)

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    Abstract

    Therapeutics based on short interfering RNAs (siRNAs), which act by inhibiting the expression of target transcripts, represent a novel class of potent and highly specific next-generation treatments for human skin diseases. Unfortunately, the intrinsic barrier properties of the skin combined with the large size and negative charge of siRNAs make epidermal delivery of these macromolecules quite challenging. To help evaluate the in vivo activity of these therapeutics and refine delivery strategies we generated an innovative reporter mouse model that predominantly expresses firefly luciferase (luc2p) in the paw epidermis - the region of murine epidermis that most closely models the tissue architecture of human skin. Combining this animal model with state-of-the-art live animal imaging techniques, we have developed a real-time in vivo analysis work-flow that has allowed us to compare and contrast the efficacies of a wide range nucleic acid-based gene silencing reagents in the skin of live animals. While inhibition was achieved with all of the reagents tested, only the commercially available "self-delivery" modified Accell-siRNAs (Dharmacon) produced potent and sustained in vivo gene silencing. Together, these findings highlight just how informative reliable reporter mouse models can be when assessing novel therapeutics in vivo. Using this work-flow, we developed a novel clinically-relevant topical formulation that facilitates non-invasive epidermal delivery of unmodified and "self-delivery" siRNAs. Remarkably, a sustained >40% luc2p inhibition was observed after two 1-hour treatments with Accell-siRNAs in our topical formulation. Importantly, our ability to successfully deliver siRNA molecules topically brings these novel RNAi-based therapeutics one-step closer to clinical use.

    Original languageEnglish
    Pages (from-to)355-362
    Number of pages8
    JournalJournal of Controlled Release: Official Journal of the Controlled Release Society, and of the Japan Society of Drug Delivery System
    Volume196
    DOIs
    Publication statusPublished - 28 Dec 2014

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    Gene Silencing
    Small Interfering RNA
    Skin
    Workflow
    Epidermis
    Firefly Luciferases
    Therapeutics
    Skin Diseases
    Nucleic Acids
    Animal Models

    Cite this

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    title = "In vivo gene silencing following non-invasive siRNA delivery into the skin using a novel topical formulation",
    abstract = "Therapeutics based on short interfering RNAs (siRNAs), which act by inhibiting the expression of target transcripts, represent a novel class of potent and highly specific next-generation treatments for human skin diseases. Unfortunately, the intrinsic barrier properties of the skin combined with the large size and negative charge of siRNAs make epidermal delivery of these macromolecules quite challenging. To help evaluate the in vivo activity of these therapeutics and refine delivery strategies we generated an innovative reporter mouse model that predominantly expresses firefly luciferase (luc2p) in the paw epidermis - the region of murine epidermis that most closely models the tissue architecture of human skin. Combining this animal model with state-of-the-art live animal imaging techniques, we have developed a real-time in vivo analysis work-flow that has allowed us to compare and contrast the efficacies of a wide range nucleic acid-based gene silencing reagents in the skin of live animals. While inhibition was achieved with all of the reagents tested, only the commercially available {"}self-delivery{"} modified Accell-siRNAs (Dharmacon) produced potent and sustained in vivo gene silencing. Together, these findings highlight just how informative reliable reporter mouse models can be when assessing novel therapeutics in vivo. Using this work-flow, we developed a novel clinically-relevant topical formulation that facilitates non-invasive epidermal delivery of unmodified and {"}self-delivery{"} siRNAs. Remarkably, a sustained >40{\%} luc2p inhibition was observed after two 1-hour treatments with Accell-siRNAs in our topical formulation. Importantly, our ability to successfully deliver siRNA molecules topically brings these novel RNAi-based therapeutics one-step closer to clinical use.",
    author = "Vikas Hegde and Hickerson, {Robyn P.} and Sitheswaran Nainamalai and Campbell, {Paul A.} and Smith, {Frances J. D.} and McLean, {W. H. Irwin} and {Leslie Pedrioli}, {Deena M.}",
    note = "Crown Copyright {\circledC} 2014. Published by Elsevier B.V. All rights reserved.",
    year = "2014",
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    doi = "10.1016/j.jconrel.2014.10.022",
    language = "English",
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    T1 - In vivo gene silencing following non-invasive siRNA delivery into the skin using a novel topical formulation

    AU - Hegde, Vikas

    AU - Hickerson, Robyn P.

    AU - Nainamalai, Sitheswaran

    AU - Campbell, Paul A.

    AU - Smith, Frances J. D.

    AU - McLean, W. H. Irwin

    AU - Leslie Pedrioli, Deena M.

    N1 - Crown Copyright © 2014. Published by Elsevier B.V. All rights reserved.

    PY - 2014/12/28

    Y1 - 2014/12/28

    N2 - Therapeutics based on short interfering RNAs (siRNAs), which act by inhibiting the expression of target transcripts, represent a novel class of potent and highly specific next-generation treatments for human skin diseases. Unfortunately, the intrinsic barrier properties of the skin combined with the large size and negative charge of siRNAs make epidermal delivery of these macromolecules quite challenging. To help evaluate the in vivo activity of these therapeutics and refine delivery strategies we generated an innovative reporter mouse model that predominantly expresses firefly luciferase (luc2p) in the paw epidermis - the region of murine epidermis that most closely models the tissue architecture of human skin. Combining this animal model with state-of-the-art live animal imaging techniques, we have developed a real-time in vivo analysis work-flow that has allowed us to compare and contrast the efficacies of a wide range nucleic acid-based gene silencing reagents in the skin of live animals. While inhibition was achieved with all of the reagents tested, only the commercially available "self-delivery" modified Accell-siRNAs (Dharmacon) produced potent and sustained in vivo gene silencing. Together, these findings highlight just how informative reliable reporter mouse models can be when assessing novel therapeutics in vivo. Using this work-flow, we developed a novel clinically-relevant topical formulation that facilitates non-invasive epidermal delivery of unmodified and "self-delivery" siRNAs. Remarkably, a sustained >40% luc2p inhibition was observed after two 1-hour treatments with Accell-siRNAs in our topical formulation. Importantly, our ability to successfully deliver siRNA molecules topically brings these novel RNAi-based therapeutics one-step closer to clinical use.

    AB - Therapeutics based on short interfering RNAs (siRNAs), which act by inhibiting the expression of target transcripts, represent a novel class of potent and highly specific next-generation treatments for human skin diseases. Unfortunately, the intrinsic barrier properties of the skin combined with the large size and negative charge of siRNAs make epidermal delivery of these macromolecules quite challenging. To help evaluate the in vivo activity of these therapeutics and refine delivery strategies we generated an innovative reporter mouse model that predominantly expresses firefly luciferase (luc2p) in the paw epidermis - the region of murine epidermis that most closely models the tissue architecture of human skin. Combining this animal model with state-of-the-art live animal imaging techniques, we have developed a real-time in vivo analysis work-flow that has allowed us to compare and contrast the efficacies of a wide range nucleic acid-based gene silencing reagents in the skin of live animals. While inhibition was achieved with all of the reagents tested, only the commercially available "self-delivery" modified Accell-siRNAs (Dharmacon) produced potent and sustained in vivo gene silencing. Together, these findings highlight just how informative reliable reporter mouse models can be when assessing novel therapeutics in vivo. Using this work-flow, we developed a novel clinically-relevant topical formulation that facilitates non-invasive epidermal delivery of unmodified and "self-delivery" siRNAs. Remarkably, a sustained >40% luc2p inhibition was observed after two 1-hour treatments with Accell-siRNAs in our topical formulation. Importantly, our ability to successfully deliver siRNA molecules topically brings these novel RNAi-based therapeutics one-step closer to clinical use.

    U2 - 10.1016/j.jconrel.2014.10.022

    DO - 10.1016/j.jconrel.2014.10.022

    M3 - Article

    C2 - 25449884

    VL - 196

    SP - 355

    EP - 362

    JO - Journal of Controlled Release: Official Journal of the Controlled Release Society, and of the Japan Society of Drug Delivery System

    JF - Journal of Controlled Release: Official Journal of the Controlled Release Society, and of the Japan Society of Drug Delivery System

    SN - 0168-3659

    ER -