TY - JOUR
T1 - RNA Aptamer Delivery through Intact Human Skin
AU - Lenn, Jon D
AU - Neil, Jessica
AU - Donahue, Christine
AU - Demock, Kellie
AU - Tibbetts, Caitlin Vestal
AU - Cote-Sierra, Javier
AU - Smith, Susan H
AU - Rubenstein, David
AU - Therrien, Jean-Philippe
AU - Pendergrast, P Shannon
AU - Killough, Jason
AU - Brown, Marc B
AU - Williams, Adrian C
N1 - This document is the Accepted Manuscript of the following article: Jon D. Lenn, et al, ‘RNA Aptamer Delivery through Intact Human Skin’, Journal of Investigative Dermatology, Vol. 138 (2): 282-290, February 2018. Under embargo until 20 September 2018.
The final, published version is available online at DOI:
https://doi.org/10.1016/j.jid.2017.07.851.
PY - 2018/2/1
Y1 - 2018/2/1
N2 - It is generally recognized that only relatively small molecular weight (typically < ∼ 500 Da) drugs can effectively permeate through intact stratum corneum. Here, we challenge this orthodoxy using a 62-nucleotide (molecular weight = 20,395 Da) RNA-based aptamer, highly specific to the human IL-23 cytokine, with picomolar activity. Results demonstrate penetration of the aptamer into freshly excised human skin using two different fluorescent labels. A dual hybridization assay quantified aptamer from the epidermis and dermis, giving levels far exceeding the cellular half maximal inhibitory concentration values (>100,000-fold), and aptamer integrity was confirmed using an oligonucleotide precipitation assay. A T helper 17 response was stimulated in freshly excised human skin resulting in significantly upregulated IL-17f, and IL-22; topical application of the IL-23 aptamer decreased both IL-17f and IL-22 by approximately 45% but did not result in significant changes to IL-23 mRNA levels, confirming that the aptamer did not globally suppress mRNA levels. This study demonstrates that very-large-molecular-weight RNA aptamers can permeate across the intact human skin barrier to therapeutically relevant levels into both the epidermis and dermis and that the skin-penetrating aptamer retains its biologically active conformational structure capable of binding to endogenous IL-23.
AB - It is generally recognized that only relatively small molecular weight (typically < ∼ 500 Da) drugs can effectively permeate through intact stratum corneum. Here, we challenge this orthodoxy using a 62-nucleotide (molecular weight = 20,395 Da) RNA-based aptamer, highly specific to the human IL-23 cytokine, with picomolar activity. Results demonstrate penetration of the aptamer into freshly excised human skin using two different fluorescent labels. A dual hybridization assay quantified aptamer from the epidermis and dermis, giving levels far exceeding the cellular half maximal inhibitory concentration values (>100,000-fold), and aptamer integrity was confirmed using an oligonucleotide precipitation assay. A T helper 17 response was stimulated in freshly excised human skin resulting in significantly upregulated IL-17f, and IL-22; topical application of the IL-23 aptamer decreased both IL-17f and IL-22 by approximately 45% but did not result in significant changes to IL-23 mRNA levels, confirming that the aptamer did not globally suppress mRNA levels. This study demonstrates that very-large-molecular-weight RNA aptamers can permeate across the intact human skin barrier to therapeutically relevant levels into both the epidermis and dermis and that the skin-penetrating aptamer retains its biologically active conformational structure capable of binding to endogenous IL-23.
UR - http://www.scopus.com/inward/record.url?scp=85041662034&partnerID=8YFLogxK
U2 - 10.1016/j.jid.2017.07.851
DO - 10.1016/j.jid.2017.07.851
M3 - Article
C2 - 28942363
SN - 0022-202X
VL - 138
SP - 282
EP - 290
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 2
ER -