-
Claire A Scott,
Vincent Plagnol,
Daniela Nitoiu,
Philip J Bland,
Diana C Blaydon,
Catherine M Chronnell,
Daniel S Poon,
David Bourn,
László Gárdos,
Andrea Császár, [......],
Malcolm Rustin,
Nigel P Burrows,
Chris Bennett, John I Harper,
Bernard Conrad,
Ishwar C Verma,
Saleem M Taibjee,
Celia Moss,
Edel A O'Toole,
David P Kelsell
Journal of Investigative Dermatology 09/2012; · 6.31 Impact Factor
-
Diana C Blaydon,
Paolo Biancheri,
Wei-Li Di,
Vincent Plagnol,
Rita M Cabral,
Matthew A Brooke,
David A van Heel,
Franz Ruschendorf,
Mark Toynbee,
Amanda Walne,
Edel A O'Toole,
Joanne E Martin,
Keith Lindley,
Tom Vulliamy,
Dominic J Abrams,
Thomas T MacDonald, John I Harper,
David P Kelsell
[show abstract]
[hide abstract]
ABSTRACT: We performed genetic and immunohistochemical studies in a sister and brother with autosomal recessive neonatal inflammatory skin and bowel lesions. The girl died suddenly at 12 years of age from parvovirus B19-associated myocarditis; her brother had mild cardiomyopathy. We identified a loss-of-function mutation in ADAM17, which encodes a disintegrin and metalloproteinase 17 (also called tumor necrosis factor α [TNF-α]-converting enzyme, or TACE), as the probable cause of this syndrome. Peripheral-blood mononuclear cells (PBMCs) obtained from the brother at 17 years of age showed high levels of lipopolysaccharide-induced production of interleukin-1β and interleukin-6 but impaired release of TNF-α. Despite repeated skin infections, this young man has led a relatively normal life. (Funded by Barts and the London Charity and the European Commission Seventh Framework Programme.).
New England Journal of Medicine 10/2011; 365(16):1502-8. · 53.30 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Gene-modified skin grafts, produced through gene transfer to human keratinocyte stem cells, offer the possibility of therapeutic benefit for inherited skin diseases. We have previously described efficient lentiviral vector-mediated gene transfer to keratinocyte stem cells and the generation of human skin grafts for the inherited skin disease, Netherton syndrome, which arises due to mutations in serine protease inhibitor Kazal-type 5 (SPINK5). Vectors incorporating an internal murine retroviral-derived promoter [spleen focus-forming virus (SFFV)] in combination with a codon-optimized SPINK5 transgene supported high levels of reconstitution and robust correction of skin architecture. Subsequent longer-term experiments have uncovered unanticipated silencing phenomena, with loss of SPINK5 gene expression over time. The inadvertent introduction of CpG sites during codon optimization appears to have rendered vectors susceptible to silencing due to methylation across the promoter-transgene boundary. Substitution of the methylation-susceptible SFFV promoter with a 572-bp minimal human involucrin promoter (INVOp), which encodes very few CpG sites, prevented repression of the SPINK5 transgene and resulted in durable and highly compartment-specific reconstitution of lympho-epithelial Kazal-type-related inhibitor (LEKTI) in human skin grafted onto immunodeficient mice. We conclude that skin grafts modified with lentiviral vectors encoding INVOp offer a suitable platform for therapeutic gene therapy in Netherton syndrome, and our experience highlights unanticipated effects of transgene codon optimization.
Human gene therapy 09/2011; 23(1):83-90. · 4.20 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Netherton syndrome (NS) is a debilitating congenital skin disorder caused by mutations in the SPINK5 gene encoding the lymphoepithelial Kazal-type-related inhibitor (LEKTI). It is characterized by defective keratinization, recurrent infections, and hypernatraemic dehydration with a mortality rate of about 10% in the first year of life. Currently, there are no curative treatments for NS. We have developed a HIV-1 based, self-inactivating lentiviral vector to express SPINK5 in keratinocytes as part of an ex-vivo gene therapy strategy for NS. High transduction efficiency was achieved in NS keratinocytes and reconstitution of LEKTI expression was confirmed in previously deficient cells. These genetically corrected keratinocytes were further tested in an in vitro organotypic culture (OTC) system and in vivo mouse/human skin engraftment model. Results showed correction of epidermal architecture in both OTCs and regenerated skin grafts. Importantly, the results from corrected skin grafts indicated that even where detectable LEKTI expression was restored to a limited numbers of cells, a wider bystander benefit occurred around these small populations. As LEKTI is a secreted protein, the genetically modified graft may provide not only an immediate local protective barrier, but also act as a source of secreted LEKTI providing a generalized benefit following ex-vivo gene therapy.
Molecular Therapy 09/2010; 19(2):408-16. · 6.87 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The autosomal recessive congenital ichthyoses are a family of related diseases, causing a severe defect in the barrier function of the epidermis. Neonates are usually born as collodion babies, but later form scales characteristic of the disease, due to a combination of thickening of the cornified layer and an increase in the production of non-polar lipids. Current treatments of choice are exfoliative creams and moisturizing agents and the use of oral retinoids. The skin condition and treatment impact significantly on quality of life and, with oral retinoids, there are potential complications associated with long-term use. A greater understanding of the mechanisms that result in scaling should lead to better directed therapies, not only for the inherited ichthyoses, but also other hyperkeratotic disorders. Using siRNA knockdown of the principle gene mutated in lamellar ichthyosis (LI), transglutaminase-1, in rat keratinocytes, we created an in vitro organotypic culture model that closely mimics the disease. Interleukin-1 alpha (IL1A) expression was increased and there was a lack of loricrin cross-linking. All LI patients tested had an increased IL1A and treatment of wild-type organotypic cultures with IL1A was sufficient to induce hyperkeratosis. Treatment of disease mimic organotypic cultures with IL-1 receptor antagonist led to a dose-dependent decrease in hyperkeratosis without a reduction in non-polar lipids in the cornified layer, which has the potential to reduce scaling without the requirement to constantly apply emollients.
Human Molecular Genetics 04/2010; 19(13):2594-605. · 7.64 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Localised scleroderma (LS) is the most common form of scleroderma seen in children, and usually presents unilaterally. Infrared thermography (IRT) and laser Doppler (LD) have both been reported to be useful in assessing the active, inflammatory stage of LS. We developed and validated a protocol using these techniques for the assessment of unilateral LS activity in children.
We investigated the spatial variability and repeatability of LD measurements from adult control forearm skin, and the inter- and intra-operator reproducibility of both LD blood flow trace analysis and IRT skin temperature analysis. Software was developed to produce overlay images of thermograms onto digital photographs of skin sites. In a group of seven adult control subjects, we established the normal range for skin temperature and LD blood flow at six standardised sites (forehead, cheek, abdomen, back, arm and leg), and measured contralateral differences in readings. In a group of 34 children with LS, we investigated the skin temperature and LD blood flow in unaffected skin at the same six sites.
In adults, physiological variability in LD blood flow and skin temperature between the two sides of the body was found to be greater than the uncertainty introduced into the measurements by (inter alia) limited intra- or inter-operator reproducibility. The cheek displayed the highest mean asymmetry in both skin temperature (0.5 degrees C) and LD blood flow (40%).
Our protocol combines IRT, LD and photography for LS assessment in children, and establishes a normal range of readings in line with other authors.
Skin Research and Technology 09/2009; 15(3):346-56. · 1.71 Impact Factor
-
Archives of Pediatrics and Adolescent Medicine 11/2007; 161(10):1001; discussion 1002. · 4.14 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Assessment of disease activity is a major challenge in the management of children with localized scleroderma. The aim of this study was to evaluate the role of laser Doppler flowmetry (LDF) in comparison with infrared thermography in the detection of scleroderma disease activity.
In 41 children with localized scleroderma, 111 lesions were assessed on 2 separate occasions, by clinical examination, LDF, and thermography. Measurements from contralateral areas of unaffected skin served as intrapatient controls, and differences in blood flow and temperature were calculated between the corresponding sites. The sensitivity and specificity to detect clinically active lesions were compared between LDF and thermography.
Seventy-five active lesions (34%) and 147 inactive lesions (66%) were identified clinically. The median relative increase in blood flow measured by LDF was +89% (range -69% to +449%) for clinically active lesions and +11% (range -46% to +302%) for clinically inactive lesions (P < 0.001). Thermography showed a median difference in temperature of +0.5 degrees C (range -0.1 degrees C to +4.1 degrees C) and +0.3 degrees C (range -1.9 degrees C to +2.7 degrees C) for clinically active lesions and clinically inactive lesions, respectively (P = 0.024). Using a cutoff level of 39% to indicate increase in blood flow, a sensitivity of 80% and specificity of 77% to detect clinically active lesions were observed; for thermography, no useful cutoff level was identified. The correlation between differences in blood flow and differences in temperature was small, but significant (r2 = 0.120, P < 0.001).
LDF is a helpful, noninvasive diagnostic technique that can be used to discriminate disease activity in children with localized scleroderma, and is more accurate than thermography for this purpose.
Arthritis & Rheumatism 11/2007; 56(10):3489-95. · 7.87 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The skin barrier (stratum corneum) is a major factor for determining the nature of immune response to antigens presented at the skin surface. Genetic abnormalities in skin barrier function are associated with allergy and atopic dermatitis, and removal of the skin barrier by tape stripping results in dominant Th2 responses to protein antigens. Langerhans cells take up antigen applied to skin from which the skin barrier has been removed; they then migrate to draining lymph nodes and initiate typical Th2 responses. These observations lead us to propose that the high frequency of allergic disease in the industrialized world might be due to lifestyle choices that result in loss of integrity of the skin barrier such as excessive washing and exfoliation of the skin.
Trends in Immunology 08/2007; 28(7):294-8. · 10.40 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Atopic dermatitis (AD) results from strong genetic and environmental interactions. AD shows genetic linkage to Chromosome 1q21. This region contains the epidermal differentiation complex (EDC), which consists of genes that form essential components of epidermal surfaces. Filaggrin (FLG) is one of these. Mutations in FLG/(R501X and 2282del4) are reported to be strongly associated with AD and to influence asthma accompanying AD. We investigated these effects in families recruited through a child with severe AD. We genotyped two panels of families, totalling 426, containing 990 affected and unaffected children. We found significant associations with AD (P=0.0001), asthma (P=0.006), and atopy (P=0.002). The FLG mutations were present in 26.7% of patients with AD, but were also present in 14.4% of children without AD. They were weakly associated with disease severity. The variants were not independently associated with asthma. The overall LOD score for genetic linkage of markers to the region was 3.57. This fell to 2.03 after accounting for the FLG mutations, indicating the presence of other genetic variants influencing AD at this locus. Our results provide further confirmation of the importance of mutations in FLG and the skin barrier in AD pathogenesis. The results indicate that investigations of other genes within the EDC should be undertaken.
Journal of Investigative Dermatology 08/2007; 127(7):1667-72. · 6.31 Impact Factor
-
The Journal of pediatrics 06/2007; 150(5):560. · 4.02 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To establish whether the prognosis of bilateral facial capillary malformation (BFCM) is worse compared with that of unilateral facial port-wine stain.
Retrospective study.
Paediatric Dermatology Department, Great Ormond Street Hospital for Children NHS Trust, a tertiary referral center for vascular anomalies.
A cohort of 350 children who presented with facial CM was seen between January 1, 1994, and June 30, 2004. Twenty-seven children with BFCM were identified. A control group of 27 children with unilateral CM was randomly selected from the total cohort.
Demographic, clinical, and radiographic characteristics were recorded and compared between the 2 groups: age at presentation, sex, distribution, extension, extrafacial lesions, glaucoma, ipsilateral leptomeningeal angiomatosis, and epilepsy. The recorded information was collected from the database of the Paediatric Dermatology Department, the hospital records, and the patients' photographs.
Compared with the 27 children with unilateral facial CM, the 27 with BFCM showed a higher frequency of association with extrafacial lesions (17 [63%] vs 6 [22%]), glaucoma (21 [78%] vs 2 [7%]), and ipsilateral leptomeningeal angiomatosis (14 [52%] vs 2 [7%]). All patients who had BFCM with bilateral and complete involvement of the ophthalmic area had ipsilateral leptomeningeal angiomatosis.
Patients with BFCM must be considered as a group with a worse prognosis compared with patients with unilateral facial CM.
Archives of Dermatology 09/2006; 142(8):994-8. · 3.89 Impact Factor
-
Archives of Dermatology 04/2006; 142(3):396-8. · 3.89 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Psoriasis and atopic dermatitis (AD) are strongly genetic and inherited as multi-factorial traits. In both diseases, linkage has been reported to chromosome 17q25. For psoriasis, the locus has been labelled PSORS2. Two peaks of association here contain the psoriasis candidate genes SLC9A3R (solute carrier family 9, isoform 3 regulatory factor), NAT9 (N-acetyltransferase superfamily), and RAPTOR (rapamycin (TOR)). We genotyped 14 of the most significantly associated single-nucleotide polymorphisms (SNPs) in these genes in a panel of 148 families (ECZ1) identified through a proband with active AD. The panel contains 350 siblings and 245 sib-pairs. Replication of positive findings was sought in a second panel, MRC-E, comprising of 278 families, 634 siblings, and 470 sib-pairs. SNP genotyping was carried out by Sequenom MassArray technology. Using family-based tests of association (transmission disequilibrium test), rs878906, in intron 3 of NAT9, was significantly associated with AD (P = 0.010) in the ECZ1 panel. In the MRC-E panel, rs895691, between the end of exon 6 of SLC9A3R1 and exon 7 of NAT9, was associated with AD (P = 0.037). These were not significant when multiple comparisons were taken into account. Haplotype analysis revealed no significant associations in either population. These results suggest that the psoriasis candidate genes do not account for previously observed linkage of the 17q25 PSORS2 locus to AD.
Journal of Investigative Dermatology 04/2006; 126(3):603-6. · 6.31 Impact Factor
-
David P Kelsell,
Elizabeth E Norgett,
Harriet Unsworth,
Muy-Teck Teh,
Thomas Cullup,
Charles A Mein,
Patricia J Dopping-Hepenstal,
Beverly A Dale,
Gianluca Tadini,
Philip Fleckman, [......],
Andrew Ilchyshyn,
Cameron T Kennedy,
Helen Goodyear,
Celia Moss,
David Paige, John I Harper,
Bryan D Young,
Irene M Leigh,
Robin A J Eady,
Edel A O'Toole
[show abstract]
[hide abstract]
ABSTRACT: Harlequin ichthyosis (HI) is the most severe and frequently lethal form of recessive congenital ichthyosis. Although defects in lipid transport, protein phosphatase activity, and differentiation have been described, the genetic basis underlying the clinical and cellular phenotypes of HI has yet to be determined. By use of single-nucleotide-polymorphism chip technology and homozygosity mapping, a common region of homozygosity was observed in five patients with HI in the chromosomal region 2q35. Sequencing of the ABCA12 gene, which maps within the minimal region defined by homozygosity mapping, revealed disease-associated mutations, including large intragenic deletions and frameshift deletions in 11 of the 12 screened individuals with HI. Since HI epidermis displays abnormal lamellar granule formation, ABCA12 may play a critical role in the formation of lamellar granules and the discharge of lipids into the intercellular spaces, which would explain the epidermal barrier defect seen in this disorder. This finding paves the way for early prenatal diagnosis. In addition, functional studies of ABCA12 will lead to a better understanding of epidermal differentiation and barrier formation.
The American Journal of Human Genetics 06/2005; 76(5):794-803. · 10.60 Impact Factor
-
Youming Zhang,
Nicholas I Leaves,
Gavin G Anderson,
Chris P Ponting,
John Broxholme,
Richard Holt,
Pauline Edser,
Sumit Bhattacharyya,
Andy Dunham,
Ian M Adcock, [......],
Gonçalo Abecasis,
Lon Cardon,
Melanie White,
John Burton,
Lucy Matthews,
Richard Mott,
Mark Ross,
Roger Cox,
Miriam F Moffatt,
William O C M Cookson
[show abstract]
[hide abstract]
ABSTRACT: Atopic or immunoglobulin E (IgE)-mediated diseases include the common disorders of asthma, atopic dermatitis and allergic rhinitis. Chromosome 13q14 shows consistent linkage to atopy and the total serum IgE concentration. We previously identified association between total serum IgE levels and a novel 13q14 microsatellite (USAT24G1; ref. 7) and have now localized the underlying quantitative-trait locus (QTL) in a comprehensive single-nucleotide polymorphism (SNP) map. We found replicated association to IgE levels that was attributed to several alleles in a single gene, PHF11. We also found association with these variants to severe clinical asthma. The gene product (PHF11) contains two PHD zinc fingers and probably regulates transcription. Distinctive splice variants were expressed in immune tissues and cells.
Nature Genetics 07/2003; 34(2):181-6. · 35.53 Impact Factor
-
Emmanuelle Bitoun,
Stéphane Chavanas,
Alan D Irvine,
Lorne Lonie,
Christine Bodemer,
Mauro Paradisi,
Dominique Hamel-Teillac,
Shin-ichi Ansai,
Yoshihiko Mitsuhashi,
Alain Taïeb,
Yves de Prost,
Giovanna Zambruno, John I Harper,
Alain Hovnanian
[show abstract]
[hide abstract]
ABSTRACT: Netherton syndrome is a severe autosomal recessive skin disorder characterized by congenital erythroderma, a specific hair-shaft abnormality, and atopic manifestations with high IgE levels. Recently, we identified SPINK5, which encodes the serine protease inhibitor Kazal-type 5 protein (LEKTI), as the defective gene in Netherton syndrome. Here we describe the intron-exon organization of the gene and characterize the SPINK5 mutations in patients from 21 families of different geographic origin, using denaturing high performance liquid chromatography and direct sequencing. We identified 18 mutations, of which 13 were novel and seven (39%) were recurrent. The majority of the mutations were clustered between exons 1-8 and exons 21-26. They comprised four nonsense mutations (22%), eight frameshift insertions or deletions (44%), and six splice-site defects (33%). All mutations predict the formation of premature termination codons. Northern blot analysis showed variable reduction of SPINK5 mutant transcript levels, suggesting variable efficiency of nonsense-mediated mRNA decay. Seven patients were homozygotes, eight were compound heterozygotes, and five were heterozygotes with only one identifiable SPINK5 mutation. Five mutations, one of which resulted in perinatal lethal disease in three families, were associated with certain ethnic groups. We also describe 45 intragenic polymorphisms in the patients studied. The clinical features of erythroderma, trichorrhexis invaginata, and atopic manifestations were present in the majority of affected individuals and ichthyosis linearis circumflexa was seen in 12 out of 24 patients. Interfamilial and intrafamilial variation in disease severity was observed, with no clear correlation between mutations and phenotype, suggesting that the degree of severity may be affected by other factors.
Journal of Investigative Dermatology 03/2002; 118(2):352-61. · 6.31 Impact Factor
-
Andrew J. Walley,
St|[eacute]|phane Chavanas,
Miriam F. Moffatt,
Robert M. Esnouf,
Baljinder Ubhi,
Robert Lawrence,
Kenny Wong,
Gon|[ccedil]|alo R Abecasis,
E. Yvonne Jones, John I. Harper,
Alain Hovnanian,
William O.C.M. Cookson
[show abstract]
[hide abstract]
ABSTRACT: Atopic dermatitis (AD) and asthma are characterized by IgE-mediated atopic (allergic) responses to common proteins (allergens), many of which are proteinases. Loci influencing atopy have been localized to a number of chromosomal regions1, including the chromosome 5q31 cytokine cluster2,
3,
4. Netherton disease is a rare recessive skin disorder in which atopy is a universal accompaniment5. The gene underlying Netherton disease (SPINK5)6 encodes a 15-domain serine proteinase inhibitor (LEKTI) which is expressed in epithelial and mucosal surfaces and in the thymus6,
7. We have identified six coding polymorphisms in SPINK5 (Table 1) and found that a Glu420Lys variant shows significant association with atopy and AD in two independent panels of families. Our results implicate a previously unrecognized pathway for the development of common allergic illnesses.
Nature Genetics. 09/2001; 29(2):175-178.
