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Single-nucleotide-specific siRNA targeting in a dominant-negative skin model.

Authors:
Robyn P Hickerson Frances J D Smith Robert E Reeves Christopher H Contag Devin Leake Sancy A Leachman Leonard M Milstone W H Irwin McLean Roger L Kaspar

J Invest Dermatol 2008 Mar 11;128(3):594-605. Epub 2007 Oct 11.

TransDerm Inc., Santa Cruz, California, USA.

RNA interference offers a novel approach for developing therapeutics for dominant-negative genetic disorders. The ability to inhibit expression of the mutant allele without affecting wild-type gene expression could be a powerful new treatment option. Targeting the single-nucleotide keratin 6a (K6a) N171K mutation responsible for the rare monogenic skin disorder pachyonychia congenita (PC), we demonstrate that small interfering RNAs (siRNAs) can potently and selectively block expression of mutant K6a. To test whether lead siRNAs could discriminate mutant mRNA in the presence of both wild-type and mutant forms, a dominant-negative PC cell culture model was developed. As predicted for a dominant-negative disease, simultaneous expression of both wild-type and mutant K6a resulted in defective keratin filament formation. Addition of mutant-specific siRNAs allowed normal filament formation, suggesting selective inhibition of mutant K6a. The effectiveness of our siRNA in skin was tested by co-delivering a firefly luciferase/mutant K6a bicistronic reporter construct and mutant-specific siRNAs to mouse footpads. Potent inhibition of the fluorescent reporter was demonstrated using the Xenogen IVIS200 in vivo imaging system. Additionally, wild type-specific siRNAs knocked down the expression of pre-existing endogenous K6a in human keratinocytes. These results suggest that efficient delivery of these "designer siRNAs" may allow effective treatment of numerous genetic disorders including PC.

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http://dx.doi.org/10.1038/sj.jid.5701060DOI Listing
March 2008

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J Cell Biol 2018 12 2;217(12):4314-4330. Epub 2018 Nov 2.

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Non-Invasive Intravital Imaging of siRNA-Mediated Mutant Keratin Gene Repression in Skin.

Authors:
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Mol Imaging Biol 2016 Feb;18(1):34-42

TransDerm Inc., 2161 Delaware Ave., Santa Cruz, CA, 95060, USA.

Purpose: Small interfering RNAs (siRNAs) specifically and potently inhibit target gene expression. Pachyonychia congenita (PC) is a skin disorder caused by mutations in genes encoding keratin (K) 6a/b, K16, and K17, resulting in faulty intermediate filaments. A siRNA targeting a single nucleotide, PC-relevant mutation inhibits K6a expression and has been evaluated in the clinic with encouraging results. Read More

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Pachyonychia congenita patients with mutations in KRT6A have more extensive disease compared with patients who have mutations in KRT16.

Authors:
K M Spaunhurst A M Hogendorf F J D Smith B Lingala M E Schwartz A Cywinska-Bernas K J Zeman J Y Tang

Br J Dermatol 2012 Apr;166(4):875-8

Department of Dermatology, Stanford University School of Medicine, Stanford, CA, USA.

Background: Pachyonychia congenita (PC) is an autosomal dominant, very rare keratin disorder caused by mutations in any of at least four genes (KRT6A, KRT6B, KRT16 or KRT17), which can lead to hypertrophic nail dystrophy and palmoplantar keratoderma, among other manifestations. Classically, patients with mutations in KRT6A and KRT16 have been grouped to the PC-1 subtype (Jadassohn-Lewandowsky type) and KRT6B and KRT17 to PC-2 (Jackson-Lawler type).

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J Invest Dermatol 2011 May 30;131(5):1029-36. Epub 2010 Dec 30.

TransDerm, Santa Cruz, California 95060, USA.

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Department of Dermatology, Huntsman Cancer Institute, University of Utah, Salt Lake City, Utah 84112-5550, USA.

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