Therapeutic siRNAs for dominant genetic skin disorders including pachyonychia congenita.

The field of science and medicine has experienced a flood of data and technology associated with the human genome project. Over 10,000 human diseases have been genetically defined, but little progress has been made with respect to the clinical application of this knowledge. A notable exception to this exists for pachyonychia congenita (PC), a rare, dominant-negative keratin disorder. The establishment of a non-profit organization, PC Project, has led to an unprecedented coalescence of patients, scientists, and physicians with a unified vision of developing novel therapeutics for PC. Utilizing the technological by-products of the human genome project, such as RNA interference (RNAi) and quantitative RT-PCR (qRT-PCR), physicians and scientists have collaborated to create a candidate siRNA therapeutic that selectively inhibits a mutant allele of KRT6A, the most commonly affected PC keratin. In vitro investigation of this siRNA demonstrates potent inhibition of the mutant allele and reversal of the cellular aggregation phenotype. In parallel, an allele-specific quantitative real-time RT-PCR assay has been developed and validated on patient callus samples in preparation for clinical trials. If clinical efficacy is ultimately demonstrated, this "first-in-skin" siRNA may herald a paradigm shift in the treatment of dominant-negative genetic disorders.

[1]  J. Rothnagel,et al.  Mutation of a type II keratin gene (K6a) in pachyonychia congenita , 1995, Nature Genetics.

[2]  C. Babinet,et al.  Introducing a Null Mutation in the Mouse K6α and K6β Genes Reveals Their Essential Structural Role in the Oral Mucosa , 2000, The Journal of cell biology.

[3]  E. Lane,et al.  Keratin 16 and keratin 17 mutations cause pachyonychia congenita , 1995, Nature Genetics.

[4]  D. Roop,et al.  Discovery of a novel murine keratin 6 (K6) isoform explains the absence of hair and nail defects in mice deficient for K6a and K6b , 2001, The Journal of cell biology.

[5]  D. Leake,et al.  Single-nucleotide-specific siRNA targeting in a dominant-negative skin model. , 2008, The Journal of investigative dermatology.

[6]  Judy Lieberman,et al.  Interfering with disease: a progress report on siRNA-based therapeutics , 2007, Nature Reviews Drug Discovery.

[7]  D. Leake,et al.  Development of therapeutic siRNAs for pachyonychia congenita. , 2008, The Journal of investigative dermatology.

[8]  T. Tuschl,et al.  Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells , 2001, Nature.

[9]  Michele De Luca,et al.  Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells , 2006, Nature Medicine.

[10]  Mouse Genome Sequencing Consortium Initial sequencing and comparative analysis of the mouse genome , 2002, Nature.

[11]  J. Schelter,et al.  Designing siRNA That Distinguish between Genes That Differ by a Single Nucleotide , 2006, PLoS genetics.

[12]  John J. Rossi,et al.  Strategies for silencing human disease using RNA interference , 2007, Nature Reviews Genetics.

[13]  D. Roop,et al.  Delayed Wound Healing in Keratin 6a Knockout Mice , 2000, Molecular and Cellular Biology.

[14]  J. Lieberman,et al.  Knocking down Disease with siRNAs , 2006, Cell.

[15]  T. Strachan,et al.  A gene for pachyonychia congenita is closely linked to the keratin gene cluster on 17q12-q21. , 1994, Journal of medical genetics.

[16]  R. Kaspar Challenges in developing therapies for rare diseases including pachyonychia congenita. , 2005, The journal of investigative dermatology. Symposium proceedings.

[17]  D. Roop,et al.  Focal Activation of a Mutant Allele Defines the Role of Stem Cells in Mosaic Skin Disorders , 2001, The Journal of cell biology.

[18]  E. Birgitte Lane,et al.  New consensus nomenclature for mammalian keratins , 2006, The Journal of cell biology.

[19]  E. Lane,et al.  Clinical and pathological features of pachyonychia congenita. , 2005, The journal of investigative dermatology. Symposium proceedings.

[20]  P. Glazer,et al.  Gene therapy for autosomal dominant disorders of keratin. , 2005, The journal of investigative dermatology. Symposium proceedings.

[21]  M. Hesse,et al.  Comprehensive analysis of keratin gene clusters in humans and rodents. , 2004, European journal of cell biology.

[22]  J. Uitto,et al.  A mutation in human keratin K6b produces a phenocopy of the K17 disorder pachyonychia congenita type 2. , 1998, Human molecular genetics.

[23]  M. Hesse,et al.  Genes for intermediate filament proteins and the draft sequence of the human genome: novel keratin genes and a surprisingly high number of pseudogenes related to keratin genes 8 and 18. , 2001, Journal of cell science.

[24]  A. Letai,et al.  Disease severity correlates with position of keratin point mutations in patients with epidermolysis bullosa simplex. , 1993, Proceedings of the National Academy of Sciences of the United States of America.