Mitsuhito Ota

Specific Filaggrin Mutations Cause Ichthyosis Vulgaris and Are Significantly Associated with Atopic Dermatitis in Japan

Mutations in the gene encoding filaggrin (FLG) have been identified as the cause of ichthyosis vulgaris (IV) and shown to be major predisposing factors for atopic dermatitis (AD). However, these studies have been mainly carried out in European populations. In early 2007, we identified two Oriental-specific FLG mutations in four Japanese families with IV and reported that filaggrin mutations were also significant predisposing factors for AD in Japan. However, the frequency of FLG mutations observed in our Japanese AD cohort (5.6%), was much lower than that seen in Europeans (up to 48%). Here, we studied a further seven Japanese families with IV and identified two additional nonsense mutations in FLG, S2889X, and S3296X. We found that more than 20% of patients in our Japanese AD case series carry FLG mutations, and there is significant statistical association between the four mutations and AD (chi(2) P=8.4 x 10(-6); heterozygote odds ratio 7.57, 95% CI 2.84-23.03). These data emphasize that skin-barrier impairment due to reduced filaggrin expression plays an important role in the pathogenesis of AD and sheds further light on the genetic architecture of atopy in Japan.

Two cases of pyoderma gangrenosum complicated with nasal septal perforation

We report two patients with pyoderma gangrenosum complicated with nasal septal perforation. An 18‐year‐old woman and a 65‐year‐old man had typical lesions of pyoderma gangrenosum on the legs that responded well to oral prednisolone. Both patients complained of mild nasal discharge, and nasal fibroscopy revealed nasal septal perforation. Biopsy of the nasal lesions showed an active inflammatory infiltrate, mainly of neutrophils. Systemic investigations failed to show any pulmonary or renal lesions of Wegener’s granulomatosis. Cytoplasmic immunofluorescent pattern antineutrophil cytoplasmic antibody was negative. In both cases, intense neutrophilic infiltration was observed not only in skin lesions but also in nasal lesions, which may indicate that the nasal lesions had a pathogenesis in common with the skin lesions.

Self-healing congenital generalized skin creases: Michelin tire baby syndrome.

REFERENCES 1. Frieden IJ, Haggstrom AN, Drolet BA, Mancini AJ, Friedlander SF, Boon L, et al. Infantile hemangiomas: current knowledge, future directions. Proceedings of a research workshop on infantile hemangiomas: April 7-9, 2005, Bethesda, Maryland, USA. Pediatr Dermatol 2005;22:383-406. 2. Hall CB, Powell KR, MacDonald NE, Gala CL, Menegus ME, Suffin SC, et al. Respiratory syncytial viral infection in children with compromised immune function. N Engl J Med 1986;315:77-81. 3. Ebbert JO, Limper AH. Respiratory syncytial virus pneumonitis in immunocompromised adults: clinical features and outcome. Respiration 2005;72:263-9. 4. American Academy of Pediatrics Committee on Infectious Diseases and Committee on Fetus and Newborn. Revised indications for the use of palivizumab and respiratory syncytial virus immune globulin intravenous for the prevention of respiratory syncytial virus infections. Pediatrics 2003;112 (6 pt 1):1442-6. 5. Light M. Respiratory syncytial virus seasonality in southeast Florida: results from three area hospitals caring for children. Pediatr Infect Dis J 2007;26(11 suppl):S55-9.

Close association of dermatitis herpetiformis and chronic tonsillitis in a Japanese patient without gluten sensitivity.

Sir, Dermatitis herpetiformis (DH) is an intensely pruritic, papulovesicular dermatosis characterized by neutrophilic microabscesses and a granular deposit of IgA in the dermal papillae. In Caucasians, many patients with DH have gluten-sensitive enteropathy (GSE), which is important to DH pathogenesis (1). HLA studies have shown the high frequency of HLA-A1 B8 DR3 DQ2 in DH and patients with coeliac disease (2). Moreover, the skin lesions and IgA deposits disappear when patients are on a strict gluten-free diet and reappear following a gluten-containing diet (3). On the other hand, in Japanese patients neither the GSE nor the particular HLA haplotype is associated with DH (4–7). Therefore, a diVerent pathogenesis of Japanese patients with DH has been expected (4, 5). We present here a Japanese patient with DH closely associated with chronic tonsillitis.

Persistent red and swollen eyelids

A 44-year-old woman presented with a 3-year history of having swollen and red eyelids. The problem was not associated with pruritus or pain. She had been previously treated with topical steroids and antibiotics but with little effect. Her past medical history was unremarkable. During physical examination, she was found to have scaly, atrophic, erythematous plaques— with the loss of eyelashes on the lateral aspects—on both lower eyelids (figure). Additionally, the scaly erythema was accompanied by hyperpigmentation on the left lateral canthus (figure). There were no other skin or mucosal lesions. Histological examination of the lesions showed follicular plugging, epidermal atrophy, liquefaction degeneration of epidermal basal cells, and dermal perivascular and periadnexal lymphocytic infiltration. Direct immunofluorescence revealed linear deposition of immunoglobulin M (IgM) at the dermoepidermal junction. She was diagnosed as having discoid lupus erythematosus (DLE); she did not meet the criteria for systemic lupus erythematosus (SLE). She was advised to avoid ultraviolet light by wearing sun glasses and sunscreen. She was also treated with oral hydroxychloroquine 300 mg/day for 16 weeks. The DLE lesions completely remitted without the improve ment of the madarosis. DLE is the most common form of chronic cutaneous lupus erythematosus, accounting for up to 80% of all cases. As many as 28% of DLE patients may go on to develop SLE. Specific risk factors for progression to SLE include female gender, widespread DLE lesions, positive antinuclear antibodies, and increased numbers of SLE criteria. Ocular complications can occur in DLE. Eyelid involvement is seen in 6% of patients. The lesion usually develops bilaterally on the lower eyelids. It may be asymptomatic; however, some patients complain of photosensitivity, burning, or swelling. Ocular DLE is commonly accompanied by facial DLE lesions—but it can be the initial or sole manifestation. DLE management begins by protecting the skin from sunlight. First-line medications are oral antimalarials such as hydroxychloroquine. Topical cortico steroids, intralesional corticosteroids, and topical cal ci neurin inhibitors may be effective for isolated manifesta tions. Early treatment is important to prevent poor cos metic results, cicatricial dysfunction, and rarely the develop ment of squamous cell carcinoma on the DLE lesion.

Multiple Hyperpigmented Macules in a Child

A 7-month-old female infant presented with a 2-month history of brown macules on the back and thighs, without prodromal erythema or trauma. Her birth history was unremarkable, and her growth and development were normal. Her family history was unremarkable. Physical examination revealed 7 sharply marginated, irregularly shaped, polymorphic hyperpigmented macules of 10 to 15 mm in diameter, without scales, on the back and thighs. The hyperpigmented macules were of orange hue, with unevenness of the pigmentation (Figure 1). Vital signs were normal. There was no lymphadenopathy or hepatosplenomegaly. Laboratory test results, including complete blood cell count, liver function tests, and renal function, were normal.

Facial Rash, Fever, and Anemia in a Newborn

A 19-day-old Japanese male infant presented with a 12-day history of fevers and facial rash. He was born by spontaneous vaginal delivery at 36 weeks’ gestational age with a birthweight of 3048 g (6.7 lb) with Apgar scores of 9 and 10. There were no complications during the prenatal and perinatal periods. His mother had no history of ringworm infection, positive IgG antibodies against rubella virus and herpes zoster virus, and serology findings negative for human immunodeficiency virus, hepatitis B virus, and syphilis. His family history was unremarkable.

Keratinocyte lipid transporter ABCA12 is highly expressed In the late stages of human epidermal development

The transfer of normal genes into somatic cells is one strategy to treat patients with genetic diseases. However, this strategy has still encounters technical problems including effi cacy of gene transfer rate and practical clinical safety. Thus, other strategies including pharmacological therapy or gene correction, are receiving increasing attention. Dystrophic epidermolysis bullosa (DEB) is caused by mutations in the COL7A1 encoding type VII collagen. This study examined the feasibility of antisense oligoribonucleotide (AON) therapy for DEB. AON was designed to induce skipping of a targeted exon containing a premature termination codon mutation, resulting in restoration of the open reading frame. We targeted exon 70 of COL7A1 since a recurrent mutation 5818delC was localized to exon 70 in Japanese DEB patients. We initially designed and synthesized two AONs to modulate splicing of exon 70 and found that one AON induced effective skipping of normal exon 70 containing the 16 amino acids. Attachment and migration analyses showed that recombinant collagen without contribution of exon 70 had similar effect to normal type VII collagen. Next, we synthesized mutation-specifi c AON by deleting cytosine at 5818. Introduction of this AON into DEB keratinocytes harboring 5818delC without expression of type VII collagen showed that the AON induced skipping of exon 70 in the abnormal 5818delC allele. Furthermore, 6.2 % of the DEB keratinocytes started to express type VII collagen in vitro after application of the mutation-specifi c AON. Injection of the AON into rat model grafted the DEB keratinocytes and fi broblasts induced detectable of type VII collagen expression at the basement membrane zone. We conclude that skipping of targeted exons using mutation-specifi c AON may show potential for future gene therapy for DEB patients.We recently reported that ABCA12 works as an epidermal keratinocyte lipid transporter, and that defective ABCA12 results in a loss of the skin lipid barrier, leading to harlequin ichthyosis (HI), one of the most devastating genodermatoses. In the present study, precise expression pattern of ABCA12 was studied in human embryonic and fetal skin of 7-22 weeks estimated gestational age (EGA) and newborn skin samples. For controls, we also studied the expression of transglutaminase 1 (TGase1) that is known to cross-link several precursor proteins in the formation of the cornifi ed cell envelope during keratinocyte terminal differentiation. In twolayered epidermis (about 6-9 weeks EGA), both ABCA12 and TGase1 were only expressed in periderm cells. In three-layered epidermis (10-13 weeks EGA), ABCA12 staining was seen not only in periderm, but also throughout the entire epidermis, while TGase1 staining was restricted to the periderm. A similar pattern was observed during four- or more-layered epidermal development (14-22 weeks EGA). In newborn skin, ABCA12 and TGase1 were seen only in the upper epidermal layers, mainly the granular layer. These staining patterns were similar to those in normal adult skin. Next, we studied ABCA12 mRNA expression in extracts of the fetal skin (at 10, 14, 15 weeks EGA). In 15 weeks EGA, the expression level of ABCA12 mRNA was signifi cantly increased compared with that in the early development (10 and 14 weeks EGA). This increasing pattern of ABCA12 mRNA expression is consistent with ABCA12 immunofl uorescent staining during human epidermal development. The unique pattern of ABCA12 expression during human epidermal development might imply severe symptoms of HI patients with ABCA12 mutations around the birth. 2006 ESDR ABSTRACTS www.Mutations in ABCA12 lead to harlequin ichthyosis and lamellar ichthyosis. The keratinocyte lipid transporter ABCA12 is a member of the ATP-binding cassette transporter family, and members of the ABCA subfamily have closely related functions as lipid transporters. Previously, we reported that the pathomechanism of harlequin ichthyosis involves the defective function of the lipid transporter ABCA12.To further elucidate the precise distribution pattern and function of ABCA12, we performed double-labeling immunofl uorescence staining for ABCA12 and for Golgi-associated or lamellar granule-associated molecules both on normal human epidermis and cultured normal human keratinocytes. We studied the precise localization of ABCA12 and other molecules using confocal laser scanning microscope. In normal human epidermis, ABCA12 was observed mainly in the granular layers with glucosylceramide (one of the major lamellar granule contents) and transglutaminase 1 (a cornifi ed cell envelope-associated keratinization marker), but not always colocalized with GM130 and TGN46 (Golgi-related molecules) that were expressed from the lower epidermis. In normal human keratinocytes cultured in high Ca++ concentration medium, ABCA12 colocalized with GM130 (a cis-Golgi- associated molecule), TGN46 (a trans-Golgi-associated molecule) and glucosylceramide. Transglutaminase 1 was restricted to the cell membrane and ABCA12 localization was within the cytoplasm distinct from transglutaminase 1 localization. The present results suggest that ABCA12 is mainly expressed in differentiated, granular layer keratinocytes with glucosylceramide and transglutaminase 1, and, at the subcellular level, ABCA12 is distributed from the cisside of the Golgi apparatus to trans-Golgi network, lamellar granules at the cell periphery. Our results suggest that ABCA12 may play an important role in lipid transport from the cis-side of the Golgi apparatus through the trans-Golgi network, to the cell periphery via lamellar granules in human epidermal granular layer keratinocytesHarlequin ichthyosis (HI) is a devastating genodermatosis that is often fatal during the neonatal period. Until the identifi cation of ABCA12-encoding a keratinocyte lipid transporter, as the causative gene for HI, prenatal diagnosis (PNDx) had been performed for more than 20 years by electron microscopic examination of fetal skin biopsy samples. We report here the fi rst case of DNA-based PNDx for HI. The proband, the fi rst child of healthy non-consanguineous French parents, showed a typical HI phenotype and died soon after birth. ABCA12 immunostaining was markedly reduced in the proband’s skin. Direct sequence analysis of ABCA12 revealed that the proband was a compound heterozygote for two novel mutations: a maternal nonsense mutation p.Ser1249X in exon 26 and a paternal missense mutation p.Arg2479Lys occurring at the last codon of exon 50. p.Ser1249X leads to an approximate 52 % truncation of the ABCA12 protein losing both ATP-binding cassette active sites. p.Arg2479Lys involves a highly conserved codon among diverse species in the second ABCA12 ATP-binding cassette. For their third pregnancy, the parents requested PNDx. Direct sequence analysis using fetal genomic DNA from amniotic fl uid cells at 17 weeks of pregnancy revealed that the fetus was a compound heterozygote for both mutations. The fetus was predicted to be affected and the parents requested the pregnancy to be terminated. The aborted fetus showed typical signs of HI. Analysis of ABCA12 transcripts of cultured keratinocytes from the abortus showed the presence of six abnormally spliced products arising from the allele carrying the missense mutation. Four of them lead to premature termination codons while the two others produced deleted proteins missing 21 and 31 amino acids in the second ATP-binding cassette. These results indicated residual expression of ABCA12. The present report paves the way for molecular PNDx of HI in the earlier stages of pregnancy