ABSTRACT
Nagashima-type palmoplantar keratosis (NPPK) is an autosomal recessive skin disorder with a high, unmet medical need that is caused by mutations in SERPINB7. Almost all NPPK patients carry the founder nonsense mutation c.796C>T (p.Arg266Ter) in the last exon of SERPINB7. Here we sought to determine whether topical nonsense-suppression (readthrough) therapy using gentamicin is applicable to NPPK. First, we demonstrated that gentamicin enhanced readthrough activity in cells transfected with SERPINB7 cDNA carrying the mutation and promoted full-length SERPINB7 protein synthesis in NPPK keratinocytes. We next conducted an investigator-blinded, randomized, bilaterally controlled compassionate use study of topical gentamicin in which five NPPK patients with c.796C>T were enrolled. Patients' self-reported improvement of hyperkeratosis was significantly greater on the gentamicin side than the control side (P = 0.0349). In two patients, hyperkeratosis was improved on the gentamicin side, as determined by a blinded-investigator assessment. These results indicate the therapeutic potential of topical gentamicin for NPPK. Unexpectedly, we also found that mutant SERPINB7 mRNAs harboring r.796c>u were degraded by nonsense-mediated mRNA decay. Furthermore, the truncated SERPINB7 protein was degraded via a proteasome-mediated pathway. These findings provide important insights into the mRNA/protein quality-control system in humans, which could be a potential therapeutic target for genetic diseases.
Subject(s)
DNA/genetics , Gentamicins/administration & dosage , Keratoderma, Palmoplantar/genetics , Mutation , Nonsense Mediated mRNA Decay/drug effects , Serpins/genetics , Administration, Topical , Codon, Nonsense , DNA Mutational Analysis , Humans , Keratinocytes/drug effects , Keratinocytes/pathology , Keratoderma, Palmoplantar/drug therapy , Keratoderma, Palmoplantar/pathology , Protein Synthesis Inhibitors/administration & dosage , Serpins/metabolismSubject(s)
Adaptor Proteins, Vesicular Transport/genetics , Etretinate/therapeutic use , Keratoderma, Palmoplantar/drug therapy , Keratoderma, Palmoplantar/genetics , Keratolytic Agents/therapeutic use , Mutation , Aged, 80 and over , Biopsy , DNA Mutational Analysis , Female , Genetic Predisposition to Disease , Humans , Immunohistochemistry , Keratoderma, Palmoplantar/diagnosis , Phenotype , Remission Induction , Treatment OutcomeSubject(s)
Lymphomatoid Papulosis/diagnosis , Skin Neoplasms/diagnosis , CD8 Antigens/metabolism , Diagnosis, Differential , Female , Humans , Ki-1 Antigen/metabolism , Lymphoma, T-Cell, Cutaneous/diagnosis , Lymphomatoid Papulosis/metabolism , Recurrence , Skin Neoplasms/metabolism , T-Lymphocytes, Cytotoxic , Young AdultABSTRACT
BACKGROUND: Atopic dermatitis (AD), Netherton syndrome (NS) and peeling skin syndrome type B (PSS) may show some clinical phenotypic overlap. Corneodesmosomes are crucial for maintaining stratum corneum integrity and the components' localization can be visualized by immunostaining tape-stripped corneocytes. In normal skin, they are detected at the cell periphery. OBJECTIVE: To determine whether AD, NS, PSS and ichthyosis vulgaris (IV) have differences in the corneodesmosomal components' distribution and corneocytes surface areas. METHODS: Corneocytes were tape-stripped from a control group (n=12) and a disease group (37 AD cases, 3 IV cases, 4 NS cases, and 3 PSS cases), and analyzed with immunofluorescent microscopy. The distribution patterns of corneodesmosomal components: desmoglein 1, corneodesmosin, and desmocollin 1 were classified into four types: peripheral, sparse diffuse, dense diffuse and partial diffuse. Corneocyte surface areas were also measured. RESULTS: The corneodesmosome staining patterns were abnormal in the disease group. Other than in the 3 PSS cases, all three components showed similar patterns in each category. In lesional AD skin, the dense diffuse pattern was prominent. A high rate of the partial diffuse pattern, loss of linear cell-cell contacts, and irregular stripping manners were unique to NS. Only in PSS was corneodesmosin staining virtually absent. The corneocyte surface areas correlated significantly with the rate of combined sparse and dense diffuse patterns of desmoglein 1. CONCLUSION: This method may be used to assess abnormally differentiated corneocytes in AD and other diseases tested. In PSS samples, tape stripping analysis may serve as a non-invasive diagnostic test.
Subject(s)
Dermatitis, Atopic/pathology , Desmosomes/pathology , Adult , Case-Control Studies , Child , Dermatitis, Atopic/metabolism , Dermatitis, Exfoliative/metabolism , Dermatitis, Exfoliative/pathology , Desmocollins/metabolism , Desmoglein 1/metabolism , Desmosomes/metabolism , Epidermis/metabolism , Epidermis/pathology , Glycoproteins/metabolism , Humans , Ichthyosis Vulgaris/metabolism , Ichthyosis Vulgaris/pathology , Intercellular Signaling Peptides and Proteins , Microscopy, Fluorescence , Netherton Syndrome/metabolism , Netherton Syndrome/pathology , Skin Diseases, Genetic/metabolism , Skin Diseases, Genetic/pathologyABSTRACT
Recessive dystrophic epidermolysis bullosa (RDEB) is an inherited skin disorder caused by mutations in the COL7A1 gene, which encodes collagen VII (COL7). Skin ulcers in RDEB patients are sometimes slow to heal. We describe here the therapeutic response of intractable skin ulcers in two patients with generalized RDEB to treatment with an allogeneic cultured dermal substitute (CDS). Skin ulcers in both patients epithelialized by 3-4 weeks after this treatment. Immunohistochemical studies demonstrated that the COL7 expression level remained reduced with respect to the control skin and that it did not differ significantly between graft-treated and untreated areas. Electron microscopy showed aberrant anchoring fibrils beneath the lamina densa of both specimens. In conclusion, CDS is a promising modality for treatment of intractable skin ulcers in patients with RDEB, even though it does not appear to increase COL7 expression.
