TY - JOUR
T1 - Use of self-delivery siRNAs to inhibit gene expression in an organotypic pachyonychia congenita model
AU - Hickerson, Robyn P.
AU - Flores, Manuel A.
AU - Leake, Devin
AU - Lara, Maria F.
AU - Contag, Christopher H.
AU - Leachman, Sancy A.
AU - Kaspar, Roger L.
N1 - Funding Information:
We thank the participating PC patients, without whom this study would not have been possible. We are indebted to Heini Ilves for expert assistance in preparation of lentiviruses and transductions. We would like to thank Peter Girling (CELLnTEC) for expert advice regarding preparation of the epidermal equivalents, Irwin McLean (University of Dundee, UK) for expert editing of the manuscript, Saleema Hassanali for technical support, and Rebecca Kaspar for proofreading, assistance in preparation of figures, and technical support. We are grateful for the generous support, collegiality, and useful suggestions from members of The International Pachyonychia Congenita Consortium (IPCC) and Pachyonychia Congenita Project, a patient advocacy group focused on finding a cure for PC. This study was funded in part by grants (RLK) from the National Institutes of Health (R43AR059474 and RC2AR058955).
PY - 2011/5
Y1 - 2011/5
N2 - Although RNA interference offers therapeutic potential for treating skin disorders, delivery hurdles have hampered clinical translation. We have recently demonstrated that high pressure, resulting from intradermal injection of large liquid volumes, facilitated nucleic acid uptake by keratinocytes in mouse skin. Furthermore, similar intradermal injections of small interfering RNA (siRNA; TD101) into pachyonychia congenita (PC) patient foot lesions resulted in improvement. Unfortunately, the intense pain associated with hypodermic needle administration to PC lesions precludes this as a viable delivery option for this disorder. To investigate siRNA uptake by keratinocytes, an organotypic epidermal model, in which pre-existing endogenous gene or reporter gene expression can be readily monitored, was used to evaluate the effectiveness of "self-delivery" siRNA (i.e., siRNA chemically modified to enhance cellular uptake). In this model system, self-delivery siRNA treatment resulted in reduction of pre-existing fluorescent reporter gene expression under conditions in which unmodified controls had little or no effect. Additionally, treatment of PC epidermal equivalents with self-delivery "TD101" siRNA resulted in marked reduction of mutant keratin 6a mRNA with little or no effect on wild-type expression. These results indicate that chemical modification of siRNA may overcome certain limitations to transdermal delivery (specifically keratinocyte uptake) and may have clinical utility for inhibition of gene expression in the skin.
AB - Although RNA interference offers therapeutic potential for treating skin disorders, delivery hurdles have hampered clinical translation. We have recently demonstrated that high pressure, resulting from intradermal injection of large liquid volumes, facilitated nucleic acid uptake by keratinocytes in mouse skin. Furthermore, similar intradermal injections of small interfering RNA (siRNA; TD101) into pachyonychia congenita (PC) patient foot lesions resulted in improvement. Unfortunately, the intense pain associated with hypodermic needle administration to PC lesions precludes this as a viable delivery option for this disorder. To investigate siRNA uptake by keratinocytes, an organotypic epidermal model, in which pre-existing endogenous gene or reporter gene expression can be readily monitored, was used to evaluate the effectiveness of "self-delivery" siRNA (i.e., siRNA chemically modified to enhance cellular uptake). In this model system, self-delivery siRNA treatment resulted in reduction of pre-existing fluorescent reporter gene expression under conditions in which unmodified controls had little or no effect. Additionally, treatment of PC epidermal equivalents with self-delivery "TD101" siRNA resulted in marked reduction of mutant keratin 6a mRNA with little or no effect on wild-type expression. These results indicate that chemical modification of siRNA may overcome certain limitations to transdermal delivery (specifically keratinocyte uptake) and may have clinical utility for inhibition of gene expression in the skin.
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U2 - 10.1038/jid.2010.426
DO - 10.1038/jid.2010.426
M3 - Article
C2 - 21248764
AN - SCOPUS:79954976090
SN - 0022-202X
VL - 131
SP - 1037
EP - 1044
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 5
ER -