Researcher Profile and Settings

Master

Affiliation (Master)

• Faculty of Medicine　Specialized Medicine　Sensory Organ Medicine

Affiliation (Master)

• Faculty of Medicine　Specialized Medicine　Sensory Organ Medicine

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Profile and Settings

Profile and Settings

• Contact Point

  natsugamed.hokudai.ac.jp

• Name (Japanese)

  Natsuga

• Name (Kana)

  Ken

• Name

  201201020285751998

Alternate Names

https://www.derm-hokudai.jp/lab-02
https://www.derm-hokudai.jp/en/lab-02

Achievement

Research Areas

• Life sciences / Dermatology

Research Experience

• 2021/04 - Today Hokkaido University
• 2021/02 - Today 北海道大学大学院医学研究院皮膚科学分野 准教授
• 2016/08 - 2021/02 Hokkaido University Hokkaido University Hospital
• 2012/09 - 2016/07 Hokkaido University Hokkaido University Hospital
• 2011/04 - 2012/08 日本学術振興会 海外特別研究員
• 2010/09 - 2012/08 Cancer Research UK Cambridge Research Institute 客員研究員
• 2010/04 - 2010/08 Hokkaido University Hokkaido University Hospital
• 2006/04 - 2007/03 Hokkaido University Hokkaido University Hospital
• 2005/04 - 2006/03 北海道がんセンター レジデント
• 2003/05 - 2005/03 Hokkaido University School of Medicine

Education

• 2007/04 - 2010/03  Hokkaido University  Graduate School of Medicine
• 1997/04 - 2003/03  Hokkaido University  School of Medicine

Awards

• 2024/08 日本研究皮膚科学会 第25回日本研究皮膚科学会賞
   

  受賞者: 夏賀 健
• 2024/08 2023年度エクセレント・ティーチャー（優秀賞） 北海道大学医学部医学科
   

  受賞者: 夏賀 健
• 2023/08 北海道大学医学部医学科 2022年度エクセレント・ティーチャー（優秀賞）
   

  受賞者: 夏賀 健
• 2022/08 Sun Pharma RISING SUN AWARD 2022 日本研究皮膚科学会
   

  受賞者: 夏賀 健
• 2022/08 2021年度エクセレント・ティーチャー（優秀賞） 北海道大学医学部医学科
   

  受賞者: 夏賀 健
• 2021/08 北海道大学医学部医学科 2020年度エクセレント・ティーチャー（優秀賞）
   

  受賞者: 夏賀 健
• 2019/08 北海道大学医学部医学科 2018年度 エクセレント・ティーチャー（最優秀賞）
   

  受賞者: 夏賀 健
• 2018/03 マルホ・高木皮膚科学振興財団 第2回高木賞
   

  受賞者: 夏賀 健
• 2014/02 第6回ロート皮膚医学研究賞
   

  受賞者: 夏賀 健
• 2013/02 第4回日本研究皮膚科学会きさらぎ賞
   

  受賞者: 夏賀 健
• 2013/02 第32回高桑榮松奨学基金奨励賞
   

  受賞者: 夏賀 健
• 2013/01 平成24年度フラテ研究奨励賞
   

  受賞者: 夏賀 健
• 2009/12 日本皮膚科学会第380回北海道地方会ベストスライド賞
   

  受賞者: 夏賀 健
• 2009/06 ESDR-JSID Young Fellow Collegiality Award
   

  受賞者: 夏賀 健
• 2007 第106回日本皮膚科学会総会ポスター賞
   

  受賞者: 夏賀 健
• 2006/06 平成17年度日本皮膚科学会北海道地方会賞
   

  受賞者: 夏賀 健
• 2005/10 日本皮膚科学会第362回北海道地方会ベストスライド賞
   

  受賞者: 夏賀 健
• 2004/12 日本皮膚科学会第359回北海道地方会ベストスライド賞
   

  受賞者: 夏賀 健
• 2004/02 日本皮膚科学会第356回北海道地方会ベストスライド賞
   

  受賞者: 夏賀 健
• 1998/12 北海道大学レーン記念賞
   

  受賞者: 夏賀 健

Published Papers

• Spatial confinement induces reciprocating migration of epidermal keratinocytes and forms triphasic epithelia

  Takuma Nohara, Junichi Kumamoto, Yosuke Mai, Mayuna Shimano, Sora Kato, Hiroyuki Kitahata, Hideki Nakamura, Shota Takashima, Mika Watanabe, Masaharu Nagayama, Tsukasa Oikawa, Hideyuki Ujiie, Ken Natsuga

  2024/11/13 

  Epithelial cells undergo epithelial–mesenchymal transition (EMT) during migration and regain their epithelial phenotype in the post-migration phase (mesenchymal–epithelial transition; MET). We established an experimental system that reproduced three-dimensional triphasic epithelia, i.e., the original epithelium, its EMT, and MET. Keratinocytes (KCs), skin epithelial cells, placed on a microporous membrane migrated through 3.0-um or larger micropores. The 3.0-um-pored membrane induced an epithelial structure with three states: stratified KCs above the membrane, KCs showing EMT within the micropores, and a new stratified epithelium under the membrane. The membrane with larger micropores failed to maintain the triphasic epithelia. Live imaging revealed that KCs moved in a reciprocating manner, with actin-rich filopodia-like KC structures extending into and out of the 3.0-um micropores, while the cells migrated unidirectionally into larger micropores. Piezo1 and keratin 6 were identified as negative modulators of KC entry to and exit from the 3.0-um micropores. These results demonstrate that non-cancerous epithelial cells migrate through confined spaces in a reciprocating manner, which might help form triphasic epithelia, recapitulating wound healing processes.
• IL-7-dependent and -independent lineages of IL-7R-dependent human T cells.

  Carlos A Arango-Franco, Masato Ogishi, Susanne Unger, Ottavia M Delmonte, Julio César Orrego, Ahmad Yatim, Margarita M Velasquez-Lopera, Andrés F Zea-Vera, Jonathan Bohlen, Marwa Chbihi, Antoine Fayand, Juan Pablo Sánchez, Julian Rojas, Yoann Seeleuthner, Tom Le Voyer, Quentin Philippot, Kathryn J Payne, Adrian Gervais, Lucia V Erazo-Borrás, Luis A Correa-Londoño, Axel Cederholm, Alejandro Gallón-Duque, Pedro Goncalves, Jean-Marc Doisne, Liran Horev, Bénédicte Charmeteau-de Muylder, Jesús Á Álvarez, Diana M Arboleda, Lizet Pérez-Zapata, Estefanía Vásquez-Echeverri, Marcela Moncada-Vélez, Juan A López, Yolanda Caicedo, Boaz Palterer, Pablo J Patiño, Carlos J Montoya, Matthieu Chaldebas, Peng Zhang, Tina Nguyen, Cindy S Ma, Mohamed Jeljeli, Juan F Alzate, Felipe Cabarcas, Taushif Khan, Darawan Rinchai, Jean-Luc Prétet, Bertrand Boisson, Nico Marr, Ruba Ibrahim, Vered Molho-Pessach, Stéphanie Boisson-Dupuis, Dimitra Kiritsi, João T Barata, Nils Landegren, Bénédicte Neven, Laurent Abel, Andrea Lisco, Vivien Béziat, Emmanuelle Jouanguy, Jacinta Bustamante, James P Di Santo, Stuart G Tangye, Luigi D Notarangelo, Rémi Cheynier, Ken Natsuga, Andrés A Arias, José Luis Franco, Klaus Warnatz, Jean-Laurent Casanova, Anne Puel

  The Journal of clinical investigation 134 (19) 2024/10/01 [Refereed]
   

  Infants with biallelic IL7R loss-of-function variants have severe combined immune deficiency (SCID) characterized by the absence of autologous T lymphocytes, but normal counts of circulating B and NK cells (T-B+NK+ SCID). We report 6 adults (aged 22 to 59 years) from 4 kindreds and 3 ancestries (Colombian, Israeli Arab, Japanese) carrying homozygous IL7 loss-of-function variants resulting in combined immunodeficiency (CID). Deep immunophenotyping revealed relatively normal counts and/or proportions of myeloid, B, NK, and innate lymphoid cells. By contrast, the patients had profound T cell lymphopenia, with low proportions of innate-like adaptive mucosal-associated invariant T and invariant NK T cells. They also had low blood counts of T cell receptor (TCR) excision circles, recent thymic emigrant T cells and naive CD4+ T cells, and low overall TCR repertoire diversity, collectively indicating impaired thymic output. The proportions of effector memory CD4+ and CD8+ T cells were high, indicating IL-7-independent homeostatic T cell proliferation in the periphery. Intriguingly, the proportions of other T cell subsets, including TCRγδ+ T cells and some TCRαβ+ T cell subsets (including Th1, Tfh, and Treg) were little affected. Peripheral CD4+ T cells displayed poor proliferation, but normal cytokine production upon stimulation with mitogens in vitro. Thus, inherited IL-7 deficiency impairs T cell development less severely and in a more subset-specific manner than IL-7R deficiency. These findings suggest that another IL-7R-binding cytokine, possibly thymic stromal lymphopoietin, governs an IL-7-independent pathway of human T cell development.
• Association between nail psoriasis and obesity: A cross-sectional study at a single institution.

  Sota Itamoto, Hajime Miyazawa, Ken Natsuga, Misako Yamaga, Hiroaki Iwata, Mika Watanabe, Hideyuki Ujiie

  The Journal of dermatology 2024/09/30 [Refereed]
  
• Type XVII collagen-specific CD4+ T cells induce bullous pemphigoid by producing IL-5.

  Norihiro Yoshimoto, Ken Muramastsu, Takamasa Ito, Miao Zheng, Kentaro Izumi, Ken Natsuga, Hiroaki Iwata, Yoshinori Hasegawa, Hideyuki Ujiie

  The Journal of investigative dermatology 2024/09/19 [Refereed]
   

  Bullous pemphigoid (BP) is an autoimmune subepidermal blistering disease caused by anti-type XVII collagen (COL17) antibodies. BP has some immunological features such as eosinophilic infiltration and the deposition of IgE autoantibodies in the skin; however, the mechanism behind such features remains largely unclear. We focused on the autoantigen-specific CD4+ T cells, which are considered to regulate immune response. We established COL17-specific CD4+ T cell lines in vitro. Wild-type mice were immunized with synthesized peptides that include a pathogenic epitope of COL17, and lymphocytes were subjected to a limiting dilution assay. We established 5 T cell lines and examined the pathogenicity by transferring them with COL17-primed B cells into Rag-2-/-/COL17-humanized mice that express human COL17 but not mouse COL17 in the skin. Notably, 3 lines induced BP-like skin changes associated with subepidermal separation and eosinophilic infiltration histologically, and the production of anti-COL17 antibodies. The other 2 lines did not induce such phenotypes. RNA-sequencing analysis revealed that Th2 cytokines, particularly IL-5, were highly expressed in the pathogenic T cell lines. Anti-IL-5 antibody administration significantly reduced the skin changes and attenuated the production of autoantibodies. Thus, the production of IL-5 is critical for COL17-specific CD4+ T cells to induce BP phenotypes in vivo.
• Patterning in stratified epithelia depends on cell-cell adhesion.

  Yosuke Mai, Yasuaki Kobayashi, Hiroyuki Kitahata, Takashi Seo, Takuma Nohara, Sota Itamoto, Shoko Mai, Junichi Kumamoto, Masaharu Nagayama, Wataru Nishie, Hideyuki Ujiie, Ken Natsuga

  Life science alliance 7 (9) 2024/09 [Refereed]
   

  Epithelia consist of proliferating and differentiating cells that often display patterned arrangements. However, the mechanism regulating these spatial arrangements remains unclear. Here, we show that cell-cell adhesion dictates multicellular patterning in stratified epithelia. When cultured keratinocytes, a type of epithelial cell in the skin, are subjected to starvation, they spontaneously develop a pattern characterized by areas of high and low cell density. Pharmacological and knockout experiments show that adherens junctions are essential for patterning, whereas the mathematical model that only considers local cell-cell adhesion as a source of attractive interactions can form regions with high/low cell density. This phenomenon, called cell-cell adhesion-induced patterning (CAIP), influences cell differentiation and proliferation through Yes-associated protein modulation. Starvation, which induces CAIP, enhances the stratification of the epithelia. These findings highlight the intrinsic self-organizing property of epithelial cells.
• Blaschkoid granulomatous pigmented purpuric dermatosis in childhood.

  George Miura, Mika Watanabe, Hideyuki Kosumi, Takuma Nohara, Ken Natsuga, Hideyuki Ujiie

  The Journal of dermatology 2024/07/17 [Refereed]
  
• Atypical manifestations of hemangiomas in epidermolysis bullosa.

  S Itamoto, K Natsuga, S Takashima, H Ujiie

  The Journal of dermatology 2024/06/14 [Refereed]
  
• Collagen 17A1 in the Urothelium Regulates Epithelial Cell Integrity and Local Immunologic Responses in Obstructive Uropathy.

  Takashi Namba, Osamu Ichii, Ken Natsuga, Teppei Nakamura, Yuki Otani, Yasuhiro Kon

  The American journal of pathology 194 (8) 1550 - 1570 2024/05/18 [Refereed]
   

  Collagen 17A1 (COL17A1), an epidermal hemidesmosome component, is ectopically induced in the urothelium of mouse and human renal pelvis (RP) in parallel with urinary tract-associated lymphoid structure development. Here, we found that COL17A1 was induced in obstructive uropathy-prone ureter of humans and cats. To ascertain its function, murine urinary organs with unilateral ureteral obstruction (UUO) were analyzed during 1 week after surgery. One day after UUO, COL17A1 expression increased in urothelial cells of RP and ureter, and was positively correlated with renal tubulointerstitial lesions. A portion of RP where the smooth muscle layer from the ureter was interrupted was sensitive to urothelium deciduation and COL17A1 induction, showing urine leaked from the RP lumen into the parenchyma. After urine stimulation, cultured immune cells expressed Cxcl2, also up-regulated in CD11b+ cells following COL17A1 stimulation. One day after UUO, CXCL2+ CD11b+ cells infiltrated the urothelium-disrupted area; however, these numbers were significantly lower in Col17a1-deficient mice. COL17A1+ urothelial cells partially co-expressed cytokeratin-14, a progenitor cell marker for urothelium, whereas Col17a1-deficient mice had lower numbers of cytokeratin-14+ cells. Gene Ontology analysis revealed that expression of epithelial- and immune-associated genes was up-regulated and down-regulated, respectively, in the ureter of Col17a1-deficient mice 4 days after UUO. Thus, COL17A1 maintains urothelium integrity by regulating urothelial cell adhesion, proliferation, and differentiation, and activates local immune responses during obstructive uropathy in mammals.
• Nail growth arrest under low body temperature during hibernation.

  Taiga Ishimoto, Hideyuki Kosumi, Ken Natsuga, Yoshifumi Yamaguchi

  The journal of physiological sciences : JPS 74 (1) 27 - 27 2024/04/27 [Refereed]
   

  Growth and differentiation are reduced or stopped during hibernation, an energy conserving strategy in harsh seasons by lowered metabolism and body temperature. However, few studies evaluated this in a same individual using a non-invasive method. In this study, we applied a non-invasive tracking method of the nail growth throughout the hibernation period in the same hibernating animals, the Syrian hamster (Mesocricetus auratus). We found that nail growth was markedly suppressed during the hibernation period but rapidly recovered by the exit from the hibernation period. Our data suggest that nail growth was arrested during deep torpor, a hypometabolic and hypothermic state, but recovered during periodic arousal, a euthermic phase. Consistent with this, nail stem cells located in the nail matrix did not exit the cell cycle in the deep torpor. Thus, hibernation stops nail growth in a body temperature-dependent manner.
• Successful treatment of multicentric Castleman's disease associated with dystrophic epidermolysis bullosa using anti-interleukin-6 receptor antibody.

  Satsuki Naruse, Shota Takashima, Yasuyuki Fujita, Hiroshi Kataoka, Nobuaki Kawamura, Ken Natsuga, Hideyuki Ujiie

  The Journal of dermatology 51 (8) e268-e269  2024/03/14 [Refereed]
  
• Variable clinical consequences of mosaicism for c.1167dupC in IKBKG in male and female patients with incontinentia pigmenti and related phenotypes.

  Roy Luister C Acos, Yi-Han Chang, Yu-Chen Lin, Dianne Katherine R Salazar-Paras, Yu Fujimura, Hajime Nakano, Eijiro Akasaka, Ken Natsuga, Bryan Edgar K Guevara, John A McGrath, Chao-Kai Hsu

  Clinical and experimental dermatology 49 (3) 298 - 301 2024/02/14 [Refereed]
  
• Transcriptomic response of peripheral blood mononuclear cells to dupilumab in a 65-year-old patient with bullous pemphigoid.

  Meng-Ling Li, Yi-Kai Hong, Yu-Chen Lin, Ken Natsuga, Hideyuki Ujiie, Kentaro Izumi, Hiroaki Iwata, Chao-Kai Hsu

  Clinical and experimental dermatology 49 (1) 73 - 74 2023/12/19 [Refereed]
  
• Dermal discoloration due to osmium tetroxide.

  Satsuki Naruse, Shota Takashima, Ken Natsuga, Hideyuki Ujiie

  Clinical toxicology (Philadelphia, Pa.) 61 (11) 1004 - 1005 2023/11 [Refereed]
   

  INTRODUCTION: Osmium tetroxide is a strong oxidizing agent. After dermal exposure to osmium tetroxide, skin discoloration and red papules can occur. We describe a patient with skin discoloration due to osmium tetroxide. CASE SUMMARY: A 25-year-old postgraduate student unintentionally exposed his hand to osmium tetroxide while working in a laboratory setting. After immediate washing, he sought medical care due to left middle finger discoloration. He reported no discomfort in the affected area. Thorough water rinsing was continued, and corticosteroid ointment was applied. IMAGES: Our patient developed dark brown pigmentation on the ventral side of the left middle finger. The pigmentation disappeared one week later. CONCLUSION: Osmium tetroxide may induce dark brown skin discoloration.
• Full-thickness Skin Grafts for Hand Contractures in an Adult Patient with Junctional Epidermolysis Bullosa: A Case Report

  Nakamura Sayaka, Saito Susumu, Ishida Yoshihiro, Kaku Yo, Natsuga Ken, Morimoto Naoki

  Journal of Plastic and Reconstructive Surgery 2 (4) 156 - 162 2023/10/27 [Refereed]
   

  Epidermolysis bullosa is a group of inherited skin fragility disorders with blister formation in the basement membrane zone. Chronic scarring after repeated blistering of the hands causes narrowing of the first web, flexion contractures of the digits, and pseudosyndactyly. Junctional epidermolysis bullosa is a type of epidermolysis bullosa that is characterized by blister formation in the lamina lucida. This condition is associated with survival into adulthood. In adult survivors, hand function might be required in order to participate in normal social activities. However, there is a lack of literature on the management of hand surgery for adult patients with junctional epidermolysis bullosa. We herein describe an adult patient with junctional epidermolysis bullosa who had pseudosyndactyly and flexion contractures in both hands. Five surgeries were performed. Contractures were released and reconstructed using split- or full-thickness skin grafts. Delayed wound healing was always observed due to epidermal necrosis in the graft. After epithelialization, a satisfactory functional outcome was obtained.
• Development of a nutritionally balanced, melt‐in‐the‐mouth chocolate for patients with epidermolysis bullosa

  Kosei Nakamura, Shota Takashima, Takuma Nohara, Mika Watanabe, Ken Natsuga, Hideyuki Ujiie

  The Journal of Dermatology 50 (12) 1640 - 1643 0385-2407 2023/09/07 [Refereed]
   

  Abstract Epidermolysis bullosa (EB) is a group of inherited blistering disorders that primarily affect the skin and mucous membranes of the digestive tract, which can lead to poor nutritional status. Dietary supplements and nutritional support methods, such as nasogastric tubes and gastrostomy, have been employed to improve the nutritional status of patients with EB; however, few foods are suitable for enjoyable eating with family and friends. Here, we introduce a nutritionally balanced, melt‐in‐the‐mouth chocolate called andew, which was specifically designed for patients with EB. The andew chocolate is nutritionally superior and melts more easily than traditional chocolates, thus it is suitable for patients with EB, who are prone to oral erosions. Patients responded more favorably to the taste and texture of andew than to those of other dietary supplements. Not only does andew provide nutritional benefits, but it also promotes enjoyable eating with family members and friends, which could positively impact patients' mental health.
• Native Autoantigen Complex Detects Pemphigoid Autoantibodies.

  Shoko Mai, Kentaro Izumi, Yosuke Mai, Ken Natsuga, Norito Ishii, Daisuke Sawamura, Franziska Schauer, Dimitra Kiritsi, Wataru Nishie, Hideyuki Ujiie

  JID innovations : skin science from molecules to population health 3 (3) 100193 - 100193 2023/05 [Refereed]
   

  Pemphigoid diseases are a group of autoimmune disorders characterized by subepidermal blistering in the skin and mucosa. Among them, mucous membrane pemphigoid (MMP) autoantibodies are characterized by targeting multiple molecules in the hemidesmosomes, including collagen XVII, laminin-332, and integrin a6/β4. Traditionally, recombinant proteins of the autoantigens have been employed to identify circulating autoantibodies by immune assays. However, developing an efficient detection system for MMP autoantibodies has been challenging because the autoantibodies have heterogeneous profiles and the antibody titers are typically low. In this study, we introduce an ELISA that takes advantage of a native autoantigen complex rather than simple recombinant proteins. We generated HaCaT keratinocytes with a DDDDK-tag knocked in at the COL17A1 locus by CRISPR/Cas9-mediated gene editing. Immunoprecipitation using the DDDDK-tag isolated a native complex that contained full-length and processed collagen XVII and integrin α6/β4. Then, we used the complex proteins to prepare an ELISA system and enrolled 55 MMP cases to validate its diagnostic performance. The sensitivity and specificity of the ELISA for detecting MMP autoantibodies were 70.9% and 86.7%, respectively, far superior to those of conventional assays. In autoimmune diseases such as MMP, in which autoantibodies target various molecules, isolating the antigen-protein complexes can help establish a diagnostic system.
• Tissue memory relies on stem cell priming in distal undamaged areas.

  Chiara Levra Levron, Mika Watanabe, Valentina Proserpio, Gabriele Piacenti, Andrea Lauria, Stefan Kaltenbach, Annalaura Tamburrini, Takuma Nohara, Francesca Anselmi, Carlotta Duval, Luca Elettrico, Daniela Donna, Laura Conti, Denis Baev, Ken Natsuga, Tzachi Hagai, Salvatore Oliviero, Giacomo Donati

  Nature cell biology 25 (5) 740 - 753 2023/05 [Refereed]
   

  Epithelial cells that participated in wound repair elicit a more efficient response to future injuries, which is believed to be locally restricted. Here we show that cell adaptation resulting from a localized tissue damage has a wide spatial impact at a scale not previously appreciated. We demonstrate that a specific stem cell population, distant from the original injury, originates long-lasting wound memory progenitors residing in their own niche. Notably, these distal memory cells have not taken part in the first healing but become intrinsically pre-activated through priming. This cell state, maintained at the chromatin and transcriptional level, leads to an enhanced wound repair that is partially recapitulated through epigenetic perturbation. Importantly wound memory has long-term harmful consequences, exacerbating tumourigenesis. Overall, we show that sub-organ-scale adaptation to injury relies on spatially organized memory-dedicated progenitors, characterized by an actionable cell state that establishes an epigenetic field cancerization and predisposes to tumour onset.
• Zonula occludens‐1 distribution and barrier functions are affected by epithelial proliferation and turnover rates

  Keisuke Imafuku, Hiroaki Iwata, Ken Natsuga, Makoto Okumura, Yasuaki Kobayashi, Hiroyuki Kitahata, Akiharu Kubo, Masaharu Nagayama, Hideyuki Ujiie

  Cell Proliferation 56 (9) e13441  0960-7722 2023/03/14 [Refereed]
   

  Zonula occludens-1 (ZO-1) is a scaffolding protein of tight junctions, which seal adjacent epithelial cells, that is also expressed in adherens junctions. The distribution pattern of ZO-1 differs among stratified squamous epithelia, including that between skin and oral buccal mucosa. However, the causes for this difference, and the mechanisms underlying ZO-1 spatial regulation, have yet to be elucidated. In this study, we showed that epithelial turnover and proliferation are associated with ZO-1 distribution in squamous epithelia. We tried to verify the regulation of ZO-1 by comparing normal skin and psoriasis, known as inflammatory skin disease with rapid turnover. We as well compared buccal mucosa and oral lichen planus, known as an inflammatory oral disease with a longer turnover interval. The imiquimod (IMQ) mouse model, often used as a psoriasis model, can promote cell proliferation. On the contrary, we peritoneally injected mice mitomycin C, which reduces cell proliferation. We examined whether IMQ and mitomycin C cause changes in the distribution and appearance of ZO-1. Human samples and mouse pharmacological models revealed that slower epithelial turnover/proliferation led to the confinement of ZO-1 to the uppermost part of squamous epithelia. In contrast, ZO-1 was widely distributed under conditions of faster cell turnover/proliferation. Cell culture experiments and mathematical modelling corroborated these ZO-1 distribution patterns. These findings demonstrate that ZO-1 distribution is affected by epithelial cell dynamics.
• Interleukin-18 as a severity marker and novel potential therapeutic target for epidermolytic ichthyosis.

  Osamu Ansai, Toshinari Miyauchi, Ryota Hayashi, Tatsuya Katsumi, Tomoki Nishiguchi, Akito Hasegawa, Satoru Shinkuma, Ken Natsuga, Toshifumi Nomura, Yutaka Shimomura, Riichiro Abe

  Clinical and experimental dermatology 48 (3) 199 - 210 2023/03/01 [Refereed]
   

  BACKGROUND: Epidermolytic ichthyosis (EI) is a major form of nonsyndromic inherited ichthyosis, characterized by erythroderma, marked hyperkeratosis and scale, bulla and erosion at birth, associated with KRT1/KRT10 mutations. The cytokine and chemokine profiles in EI are poorly understood, and specific treatment options have not been established. AIM: To explore novel biomarkers and therapeutic targets in patients with EI. METHODS: We analysed cytokine levels in serum and skin samples from 10 patients with inherited ichthyosis, including seven patients with EI. Wild-type and mutant KRT1 constructs were established and transfected into HaCaT cells, an immortalized keratinocyte cell line, for in vitro immunoblotting and immunocytochemistry analyses. RESULTS: Multiplex cytokine/chemokine analysis revealed that 10 cytokines/chemokines [interleukin (IL)-1β, IL-4, IL-17A, IL-16, IL-18, IL-1 receptor-α, macrophage colony-stimulating factor, interferon-α2, basic fibroblast growth factor and monocyte chemotactic protein-3] were significantly increased in patients with EI. Furthermore, IL-18 levels were significantly higher in patients with EI [n = 7; 2714.1 (1438.0) pg mL-1] than in healthy controls [n = 11; 218.4 (28.4) pg mL-1, P < 0.01]. Immunohistochemical analyses showed that IL-18 expression was elevated in skin samples from patients with EI. Serum IL-18 levels correlated with the severity of ichthyosis, as measured by the Ichthyosis Scoring System. Immunoblotting analysis revealed that mature IL-18 levels were increased in the supernatant of mutant KRT1 expressing HaCaT cells. Additionally, these cells showed NLRP3 aggregation in the cytoplasm and ASC clustered around mutant keratin aggregations. These findings suggest that mutant keratin might promote the activation of the NLRP3 inflammasome and its downstream caspase-1-mediated IL-18 release in keratinocytes from patients with EI. CONCLUSIONS: Our results suggest that serum IL-18 is a severity marker released from the skin of patients with EI. Blockade of IL-18 may be a useful novel therapeutic option for patients with EI.
• A case of epidermolysis bullosa simplex-Ogna with nail involvement.

  Shingo Takei, Ryota Hayashi, Tatsuya Katsumi, Osamu Ansai, Akari Sakai, Ken Natsuga, Riichiro Abe

  The Journal of dermatology 50 (6) e177-e178  2022/12/30 [Refereed]
  
• Mutational analysis of epidermolysis bullosa in Taiwan by whole-exome sequencing complemented by RNA sequencing: a series of 77 patients.

  Wei-Ting Tu, Ping-Chen Hou, Peng-Chieh Chen, Wan-Rung Chen, Hsin-Yu Huang, Jing-Yu Wang, Yi-Ting Huang, Yi-Huei Wu, Chun-Lin Su, Yen-An Tang, Hiroaki Iwata, Ken Natsuga, Sheau-Chiou Chao, H Sunny Sun, Ming-Jer Tang, Julia Yu-Yun Lee, John A McGrath, Chao-Kai Hsu

  Orphanet journal of rare diseases 17 (1) 451 - 451 2022/12/28 [Refereed]
   

  BACKGROUND: Epidermolysis bullosa (EB) is a heterogeneous group of hereditary skin diseases characterized by skin fragility. Primary data on Taiwanese population remain scarce. METHODS: We gathered clinical information from EB patients at National Cheng Kung University Hospital from January, 2012, to June, 2021. Diagnostic tests including transmission electron microscopy, immunofluorescence studies, and whole-exome sequencing (WES) were performed. The pathogenicity of novel splice-site mutations was determined through reverse transcriptase-PCR of skin mRNA followed by Sanger and/or RNA sequencing. RESULTS: Seventy-seven EB patients from 45 families were included: 19 EB simplex, six junctional EB, and 52 dystrophic EB. Pathogenic variants were identified in 37 of 38 families (97.4%), in which WES was used as a first-line tool for mutational analysis; RNA sequencing determined pathogenic variants in the remaining one family. A total of 60 mutations in EB-related genes were identified, including 22 novel mutations. The mutations involved KRT5, KRT14, PLEC, COL17A1, LAMB3, LAMA3, ITGB4, and COL7A1. Over one-quarter of DEB patients had EB pruriginosa. CONCLUSIONS: The distinct clinical presentation and molecular pathology of EB in Taiwan expand our understanding of this disorder. WES was an effective first-line diagnostic tool for identifying EB-associated variants. RNA sequencing complemented WES when multiple potentially pathogenic splice-site mutations were found.
• Acquired perforating dermatosis induced by necitumumab.

  A Kimura, H Kosumi, K Natsuga, T Goda, H Ujiie

  Journal of the European Academy of Dermatology and Venereology : JEADV 36 (10) e822-e823  2022/10 [Refereed]
  
• Apremilast for lithium-associated psoriasis

  Joohyung Youh, Hajime Miyazawa, Hiroaki Iwata, Ken Natsuga, Hideyuki Ujiie

  Dermatologica Sinica 1027-8117 2022/07 [Refereed]
  
• Collagen XVII deficiency alters epidermal patterning.

  Yunan Wang, Hiroyuki Kitahata, Hideyuki Kosumi, Mika Watanabe, Yu Fujimura, Shota Takashima, Shin-Ichi Osada, Tomonori Hirose, Wataru Nishie, Masaharu Nagayama, Hiroshi Shimizu, Ken Natsuga

  Laboratory investigation; a journal of technical methods and pathology 102 (6) 581 - 588 2022/06 [Refereed]
   

  Vertebrates exhibit patterned epidermis, exemplified by scales/interscales in mice tails and grooves/ridges on the human skin surface (microtopography). Although the role of spatiotemporal regulation of stem cells (SCs) has been implicated in this process, the mechanism underlying the development of such epidermal patterns is poorly understood. Here, we show that collagen XVII (COL17), a niche for epidermal SCs, helps stabilize epidermal patterns. Gene knockout and rescue experiments revealed that COL17 maintains the width of the murine tail scale epidermis independently of epidermal cell polarity. Skin regeneration after wounding was associated with slender scale epidermis, which was alleviated by overexpression of human COL17. COL17-negative skin in human junctional epidermolysis bullosa showed a distinct epidermal pattern from COL17-positive skin that resulted from revertant mosaicism. These results demonstrate that COL17 contributes to defining mouse tail scale shapes and human skin microtopography. Our study sheds light on the role of the SC niche in tissue pattern formation.
• Wnt/β-Catenin Signaling Stabilizes Hemidesmosomes in Keratinocytes.

  Hideyuki Kosumi, Mika Watanabe, Satoru Shinkuma, Takuma Nohara, Yu Fujimura, Tadasuke Tsukiyama, Giacomo Donati, Hiroaki Iwata, Hideki Nakamura, Hideyuki Ujiie, Ken Natsuga

  The Journal of investigative dermatology 142 (6) 1576 - 1586 2022/06 [Refereed]
   

  Hemidesmosomes (HDs) are adhesion complexes that promote epithelial-stromal attachment in stratified and complex epithelia, including the epidermis. In various biological processes, such as differentiation and migration of epidermal keratinocytes during wound healing or carcinoma invasion, quick assembly and disassembly of HDs are prerequisites. In this study, we show that inhibition of Wnt/β-catenin signaling disturbs HD organization in keratinocytes. Screening with inhibitors identified the depletion of HD components and HD-like structures through Wnt inhibition, but keratinocyte differentiation was not affected. Wnt inhibition significantly diminished plectin and type XVII collagen expression in the basal side of Wnt-inhibited cells and the dermo-epidermal junction of the Wnt-inactive murine basal epidermis. Similar to Wnt inhibition, PLEC-knockout cells or cells with plectin-type XVII collagen binding defects showed type XVII collagen reduction in the basal side of the cells, implying the possible involvement of Wnt/β-catenin signaling in HD assembly. Atypical protein kinase C inhibition ameliorated the phenotypes of Wnt-inhibited cells. These findings show that Wnt/β-catenin signaling regulates the localization of HD components in keratinocytes and that the atypical protein kinase C pathway is involved in Wnt inhibition‒induced HD disarrangement. Our study suggests that the Wnt signaling pathway could be a potential therapeutic target for treating HD-defective diseases, such as epidermolysis bullosa.
• Cas9-guided haplotyping of three truncation variants in autosomal recessive disease.

  Ken Natsuga, Yoshikazu Furuta, Shota Takashima, Takuma Nohara, Hsin-Yu Huang, Satoru Shinkuma, Hideki Nakamura, Yousuke Katsuda, Hideaki Higashi, Chao-Kai Hsu, Satoshi Fukushima, Hideyuki Ujiie

  Human mutation 43 (7) 877 - 881 2022/04/21 [Refereed]
   

  An autosomal recessive disease is caused by biallelic loss-of-function mutations. However, when more than two disease-causing variants are found in a patient's gene, it is challenging to determine which two of the variants are responsible for the disease phenotype. Here, to decipher the pathogenic variants by precise haplotyping, we applied nanopore Cas9-targeted sequencing (nCATS) to three truncation COL7A1 variants detected in a patient with recessive dystrophic epidermolysis bullosa (EB). The distance between the most 5' and 3' variants was approximately 19 kb at the level of genomic DNA. nCATS successfully demonstrated that the most 5' and 3' variants were located in one allele while the variant in between was located in the other allele. Interestingly, the proband's mother, who was phenotypically intact, was heterozygous for the allele that harbored the two truncation variants, which could otherwise be misinterpreted as those of typical recessive dystrophic EB. Our study highlights the usefulness of nCATS as a tool to determine haplotypes of complicated genetic cases. Haplotyping of multiple variants in a gene can determine which variant should be therapeutically targeted when nucleotide-specific gene therapy is applied.
• Detection of revertant mosaicism in epidermolysis bullosa through Cas9-targeted long-read sequencing.

  Ken Natsuga, Yoshikazu Furuta, Shota Takashima, Takuma Nohara, Hideyuki Kosumi, Yosuke Mai, Hideaki Higashi, Hideyuki Ujiie

  Human mutation 43 (4) 529 - 536 2022/04 [Refereed]
   

  Revertant mosaicism (RM) is a phenomenon in which inherited mutations are spontaneously corrected in somatic cells. RM occurs in some congenital skin diseases, but genetic validation of RM in clinically revertant skin has been challenging, especially when homologous recombination (HR) is responsible for RM. Here, we introduce nanopore Cas9-targeted sequencing (nCATS) for identifying HR in clinically revertant skin. We took advantage of compound heterozygous COL7A1 mutations in a patient with recessive dystrophic epidermolysis bullosa who showed revertant skin spots. Cas9-mediated enrichment of genomic DNA (gDNA) covering the two mutation sites (>8 kb) in COL7A1 and subsequent MinION sequencing successfully detected intragenic crossover in the epidermis of the clinically revertant skin. This method enables the discernment of haplotypes of up to a few tens of kilobases of gDNA. Moreover, it is devoid of polymerase chain reaction amplification, which can technically induce recombination. We, therefore, propose that nCATS is a powerful tool for understanding complicated gene modifications, including RM.
• New insight of itch mediators and proinflammatory cytokines in epidermolysis bullosa

  Hong Ha Nguyen, Satoru Shinkuma, Ryota Hayashi, Tatsuya Katsumi, Tomoki Nishiguchi, Ken Natsuga, Yasuyuki Fujita, Riichiro Abe

  Journal of Cutaneous Immunology and Allergy Frontiers Media SA 5 (3) 78 - 87 2574-4593 2022/02/07 [Refereed]
   

  AbstractObjectivesEpidermolysis bullosa (EB) is a hereditary disorder characterized by mechanical stress‐induced blistering. The presence of extracutaneous complications such as cardiomyopathy and renal disease observed in severe EB subtypes and the fact that pruritus is a common symptom across all EB subtypes indicate that EB is not only a skin fragility disease but also a systemic inflammatory disorder. Our study aims to elucidate the basis of the systemic inflammation seen in EB patients.MethodsWe analyzed serum samples of 20 EB patients by Luminex bead‐based cytokine assays and enzyme‐linked immunosorbent assays.ResultsThe serum levels of sIL‐2R, IL‐6, HGF, M‐CSF, SCGF‐β, IL‐8, IL‐16, IFN‐γ, MIF, MIP‐1α, and thymic stromal lymphopoietin (TSLP) (p < .01, p < .01, p < .01, p < .01, p < .01, p < .05, p < .05, p < .05, p < .05, p < .05, and p = .01, respectively) were found to be significantly elevated in EB patients, whereas TNF‐β (p < .01) was decreased. Th2 cytokines including IL‐4, IL‐5, and IL‐13 were not elevated in EB patients. In contrast, TSLP was significantly increased, and IL‐31 and oncostatin M were not statistically significant but tended to be higher in the patients than in healthy controls. Among proinflammatory cytokines such as IL‐1β, IL‐6, and TNF‐α, IL‐6 was elevated in EB patients.ConclusionsThe imbalance of several itch mediators and proinflammatory cytokines was identified. Biologics targeting the cytokines found to be elevated in the sera of patients is considered as a beneficial treatment option for EB.
• Prevention of chemotherapy-induced hair loss by peroxisome proliferator-activated receptor-γ modulation.

  K Natsuga

  The British journal of dermatology 186 (1) 14 - 15 2022/01 [Refereed][Invited]
  
• IgA nephropathy preceded by erythroderma with eosinophilia.

  Mina Takatsu, Ken Natsuga, Fumihiko Hattanda, Hideyuki Ujiie

  European journal of dermatology : EJD 32 (1) 127 - 129 2022/01/01 [Refereed]
  
• Case of inherited epidermolysis bullosa simplex with KLHL24 gene mutation in Japan.

  Tomoko Miyake, Ken Natsuga, Takatsune Umayahara, Seiko Naito, Junko Yoshimoto, Akemi Senoo, Han-Tang Wang, Chao-Kai Hsu, Osamu Yamasaki, Shin Morizane

  The Journal of dermatology 49 (1) e24-e25  2022/01 [Refereed]
  
• Current topics in Epidermolysis bullosa: Pathophysiology and therapeutic challenges.

  Ken Natsuga, Satoru Shinkuma, Chao-Kai Hsu, Yasuyuki Fujita, Akira Ishiko, Katsuto Tamai, John A McGrath

  Journal of dermatological science 104 (3) 164 - 176 2021/12 [Refereed][Invited]
   

  Epidermolysis bullosa (EB) is a group of inherited skin and mucosal fragility disorders resulting from mutations in genes encoding basement membrane zone (BMZ) components or proteins that maintain the integrity of BMZ and adjacent keratinocytes. More than 30 years have passed since the first causative gene for EB was identified, and over 40 genes are now known to be responsible for the protean collection of mechanobullous diseases included under the umbrella term of EB. Through the elucidation of disease mechanisms using human skin samples, animal models, and cultured cells, we have now reached the stage of developing more effective therapeutics for EB. This review will initially focus on what is known about blister wound healing in EB, since recent and emerging basic science data are very relevant to clinical translation and therapeutic strategies for patients. We then place these studies in the context of the latest information on gene therapy, read-through therapy, and cell therapy that provide optimism for improved clinical management of people living with EB.
• Unilateral Naevoid Telangiectasia Associated with Oral Contraceptive.

  Takuma Nohara, Ken Natsuga, Atsushi Yasuoka, Hideyuki Ujiie

  Acta dermato-venereologica 101 (11) adv00595  2021/11/17 [Refereed]
  
• Complexity of Transcriptional and Translational Interference of Laminin-332 Subunits in Junctional Epidermolysis Bullosa with LAMB3 Mutations.

  Ping-Chen Hou, Ken Natsuga, Wei-Ting Tu, Hsin-Yu Huang, Brandon Chen, Liang-Yu Chen, Wan-Rung Chen, Yi-Kai Hong, Yen-An Tang, Julia Yu-Yun Lee, Peng-Chieh Chen, H Sunny Sun, John A McGrath, Chao-Kai Hsu

  Acta dermato-venereologica 101 (8) adv00522  2021/08/24 [Refereed]
  
• Successful kidney transplantation in a patient with neonatal-onset ILNEB.

  Takayuki Okamoto, Akie Nakamura, Asako Hayashi, Takeshi Yamaguchi, Yayoi Ogawa, Ken Natsuga, Kumiko Yanagi, Kiyohiko Hotta

  Pediatric transplantation 25 (5) e13971  2021/08 [Refereed]
   

  BACKGROUND: ILNEB constitute an autosomal recessive disorder caused by homozygous or compound heterozygous mutation of the gene for the ITGA3. To date, 8 ILNEB patients have been reported, but all 6 neonatal-onset ILNEB patients suffered early death within 2 years. The most common cause of death among previously reported ILNEB patients was exacerbation of the respiratory condition. METHODS: In this study, we describe a case of ILNEB with neonatal onset in a female patient and the genetic and histopathological testing performed. RESULTS: Our patient had a compound heterozygous mutation in ITGA3. Compared to previously reported patients, this patient exhibited milder clinical and histopathological characteristics. After experiencing a life-threatening respiratory infection at 8 months old, the patient started periodic subcutaneous immunoglobulin treatment once every 1-2 weeks for nephrotic-range proteinuria-induced secondary hypogammaglobulinemia. At the age of 3 years, proteinuria gradually increased with severe edema despite strict internal management. Therefore, our patient underwent unilateral nephrectomy and insertion of a peritoneal dialysis catheter followed by another unilateral nephrectomy. One month later, she underwent an ABO-compatible living-donor kidney transplantation at the age of 4 years. CONCLUSIONS: Our patient is a neonatal-onset ILNEB patient who survived for more than 2 years and underwent successful kidney transplantation.
• The CD44/COL17A1 pathway promotes the formation of multilayered, transformed epithelia.

  Kei Kozawa, Miho Sekai, Kenji Ohba, Shoko Ito, Hiroaki Sako, Takeshi Maruyama, Mai Kakeno, Takanobu Shirai, Keisuke Kuromiya, Tomoko Kamasaki, Koki Kohashi, Shinya Tanaka, Susumu Ishikawa, Nanami Sato, Shota Asano, Hironori Suzuki, Nobuyuki Tanimura, Yohei Mukai, Noriko Gotoh, Mishie Tanino, Shinya Tanaka, Ken Natsuga, Tomoyoshi Soga, Tomonori Nakamura, Yukihiro Yabuta, Mitinori Saitou, Takahiro Ito, Kenkyo Matsuura, Makoto Tsunoda, Toyone Kikumori, Tadashi Iida, Yasuyuki Mizutani, Yuki Miyai, Kozo Kaibuchi, Atsushi Enomoto, Yasuyuki Fujita

  Current biology : CB 31 (14) 3086 - 3097 2021/07/26 [Refereed]
   

  At the early stage of cancer development, oncogenic mutations often cause multilayered epithelial structures. However, the underlying molecular mechanism still remains enigmatic. By performing a series of screenings targeting plasma membrane proteins, we have found that collagen XVII (COL17A1) and CD44 accumulate in RasV12-, Src-, or ErbB2-transformed epithelial cells. In addition, the expression of COL17A1 and CD44 is also regulated by cell density and upon apical cell extrusion. We further demonstrate that the expression of COL17A1 and CD44 is profoundly upregulated at the upper layers of multilayered, transformed epithelia in vitro and in vivo. The accumulated COL17A1 and CD44 suppress mitochondrial membrane potential and reactive oxygen species (ROS) production. The diminished intracellular ROS level then promotes resistance against ferroptosis-mediated cell death upon cell extrusion, thereby positively regulating the formation of multilayered structures. To further understand the functional role of COL17A1, we performed comprehensive metabolome analysis and compared intracellular metabolites between RasV12 and COL17A1-knockout RasV12 cells. The data imply that COL17A1 regulates the metabolic pathway from the GABA shunt to mitochondrial complex I through succinate, thereby suppressing the ROS production. Moreover, we demonstrate that CD44 regulates membrane accumulation of COL17A1 in multilayered structures. These results suggest that CD44 and COL17A1 are crucial regulators for the clonal expansion of transformed cells within multilayered epithelia, thus being potential targets for early diagnosis and preventive treatment for precancerous lesions.
• Hair follicle stem cell progeny heal blisters while pausing skin development.

  Yu Fujimura, Mika Watanabe, Kota Ohno, Yasuaki Kobayashi, Shota Takashima, Hideki Nakamura, Hideyuki Kosumi, Yunan Wang, Yosuke Mai, Andrea Lauria, Valentina Proserpio, Hideyuki Ujiie, Hiroaki Iwata, Wataru Nishie, Masaharu Nagayama, Salvatore Oliviero, Giacomo Donati, Hiroshi Shimizu, Ken Natsuga

  EMBO reports 22 (7) e50882  2021/07/05 [Refereed]
   

  Injury in adult tissue generally reactivates developmental programs to foster regeneration, but it is not known whether this paradigm applies to growing tissue. Here, by employing blisters, we show that epidermal wounds heal at the expense of skin development. The regenerated epidermis suppresses the expression of tissue morphogenesis genes accompanied by delayed hair follicle (HF) growth. Lineage tracing experiments, cell proliferation dynamics, and mathematical modeling reveal that the progeny of HF junctional zone stem cells, which undergo a morphological transformation, repair the blisters while not promoting HF development. In contrast, the contribution of interfollicular stem cell progeny to blister healing is small. These findings demonstrate that HF development can be sacrificed for the sake of epidermal wound regeneration. Our study elucidates the key cellular mechanism of wound healing in skin blistering diseases.
• A computational model of the epidermis with the deformable dermis and its application to skin diseases.

  Kota Ohno, Yasuaki Kobayashi, Masaaki Uesaka, Takeshi Gotoda, Mitsuhiro Denda, Hideyuki Kosumi, Mika Watanabe, Ken Natsuga, Masaharu Nagayama

  Scientific reports 11 (1) 13234 - 13234 2021/06/24 [Refereed]
   

  The skin barrier is provided by the organized multi-layer structure of epidermal cells, which is dynamically maintained by a continuous supply of cells from the basal layer. The epidermal homeostasis can be disrupted by various skin diseases, which often cause morphological changes not only in the epidermis but in the dermis. We present a three-dimensional agent-based computational model of the epidermis that takes into account the deformability of the dermis. Our model can produce a stable epidermal structure with well-organized layers. We show that its stability depends on the cell supply rate from the basal layer. Modeling the morphological change of the dermis also enables us to investigate how the stiffness of the dermis affects the structure and barrier functions of the epidermis. Besides, we show that our model can simulate the formation of a corn (clavus) by assuming hyperproliferation and rapid differentiation. We also provide experimental data for human corn, which supports the model assumptions and the simulation result.
• Altered replication stress response due to CARD14 mutations promotes recombination-induced revertant mosaicism.

  Toshinari Miyauchi, Shotaro Suzuki, Masae Takeda, Jin Teng Peh, Masayuki Aiba, Ken Natsuga, Yasuyuki Fujita, Takuya Takeichi, Taiko Sakamoto, Masashi Akiyama, Hiroshi Shimizu, Toshifumi Nomura

  American journal of human genetics 108 (6) 1026 - 1039 2021/06/03 [Refereed]
   

  Revertant mosaicism, or "natural gene therapy," refers to the spontaneous in vivo reversion of an inherited mutation in a somatic cell. Only approximately 50 human genetic disorders exhibit revertant mosaicism, implicating a distinctive role played by mutant proteins in somatic correction of a pathogenic germline mutation. However, the process by which mutant proteins induce somatic genetic reversion in these diseases remains unknown. Here we show that heterozygous pathogenic CARD14 mutations causing autoinflammatory skin diseases, including psoriasis and pityriasis rubra pilaris, are repaired mainly via homologous recombination. Rather than altering the DNA damage response to exogenous stimuli, such as X-irradiation or etoposide treatment, mutant CARD14 increased DNA double-strand breaks under conditions of replication stress. Furthermore, mutant CARD14 suppressed new origin firings without promoting crossover events in the replication stress state. Together, these results suggest that mutant CARD14 alters the replication stress response and preferentially drives break-induced replication (BIR), which is generally suppressed in eukaryotes. Our results highlight the involvement of BIR in reversion events, thus revealing a previously undescribed role of BIR that could potentially be exploited to develop therapeutics for currently intractable genetic diseases.
• Zonula occludens-1 demonstrates a unique appearance in buccal mucosa over several layers.

  Keisuke Imafuku, Mayumi Kamaguchi, Ken Natsuga, Hideki Nakamura, Hiroshi Shimizu, Hiroaki Iwata

  Cell and tissue research 384 (3) 691 - 702 2021/06 [Refereed]
   

  Tight junctions (TJs) firmly seal epithelial cells and are key players in the epithelial barrier. TJs consist of several proteins, including those of the transmembrane claudin family and the scaffold zonula occludens (ZO) family. Epithelial tissues are exposed to different conditions: to air in the stratified epithelium of the skin and to liquids in the monolayer of the intestine. The TJs in stratified oral mucosal epithelium have remained insufficiently elucidated in terms of distributions, appearances and barrier functions of TJ proteins in normal buccal mucosa. We investigated these and ZO-1 and claudin-1 were found to be expressed in the top third and in the bottom three quarters of the mucosal epithelium. ZO-1 in the buccal mucosa was found to have an irregular linear appearance. ZO-1 in the buccal mucosa continuously existed in several layers. Electron microscopy revealed the buccal mucosa to have kissing points. In a biotin permeation assay that sought to investigate inside-outside barrier function, the biotin tracer penetrated several ZO-1 layers but did not pass through all the ZO-1 layers. We found that the oral mucosal cell knockdown of TJP1 or CLDN1 resulted in decreases of TER but no significant change in FITC-dextran leakage. Our results suggest that the distribution and appearance of ZO-1 in the buccal mucosa differ from those in the skin. We were unable to prove barrier function in this study but we did show barrier function against small molecules in vivo and against ions in vitro.
• A case of non-bullous pemphigoid induced by IgG4 autoantibodies targeting BP230.

  N Yoshimoto, S Takashima, T Kawamura, E Inamura, T Sugai, I Ujiie, K Izumi, K Natsuga, W Nishie, H Shimizu, H Ujiie

  Journal of the European Academy of Dermatology and Venereology : JEADV 35 (4) e282-e285  2021/04 [Refereed]
  
• Intravenous allogeneic multilineage-differentiating stress-enduring cells in adults with dystrophic epidermolysis bullosa: a phase 1/2 open-label study.

  Y Fujita, T Nohara, S Takashima, K Natsuga, M Adachi, K Yoshida, S Shinkuma, T Takeichi, H Nakamura, O Wada, M Akiyama, A Ishiko, H Shimizu

  Journal of the European Academy of Dermatology and Venereology : JEADV 2021/03/03 [Refereed]
  
• A case of mucous membrane pemphigoid with anti-BP230 autoantibodies alone.

  Norihiro Yoshimoto, Inkin Ujiie, Emi Inamura, Ken Natsuga, Wataru Nishie, Hiroshi Shimizu, Hideyuki Ujiie

  International journal of dermatology 60 (3) e92-e94  2021/03 [Refereed]
  
• Five novel mutations in SASH1 contribute to lentiginous phenotypes in Japanese families.

  Yuta Araki, Ken Okamura, Toru Saito, Kazuhiko Matsumoto, Ken Natsuga, Junko Nishimoto, Yoko Funasaka, Yaei Togawa, Tamio Suzuki

  Pigment cell & melanoma research 34 (2) 174 - 178 2021/03 [Refereed]
   

  SASH1 has been reported as a causal gene of lentiginous phenotypes with and without heredity, including an autosomal dominant type characterized by lentigines predominantly on sun-exposed areas such as the face and limbs. Recently, cases of dyschromatosis with SASH1 mutations have been reported worldwide; however, only one case has been reported from Japan. Here, we analyzed six Japanese patients who characteristically showed many lentigines on sun-exposed areas, using next-generation sequencing. We identified five novel heterozygous mutations in SASH1 (p.I586M, p.S531Y, p.R644W, p.T525R, and p.S516I) in our patients and their families. The p.R644W substitution identified in two unrelated families was the first mutation located in the sterile alpha motif 1 (SAM1) domain. The degree and location of the lentigines were variable across individuals, even if they shared the same SASH1 mutation. All mutations were predicted to be deleterious by six different algorithms used to evaluate the functional impact of a variation. In addition, immunohistopathological findings and RNA sequencing results suggested that SASH1 mutations were associated with an increase in the number of melanocytes, acceleration of melanogenesis, and upregulated hair keratin expression.
• Dominance of Methicillin-Resistant Staphylococcus aureus in a Japanese Infant with Recessive Dystrophic Epidermolysis Bullosa.

  Makoto Kondo, Shota Takashima, Hiroyuki Goto, Koji Habe, Ken Natsuga, Keiichi Yamanaka

  Case reports in dermatology 13 (2) 278 - 281 2021 [Refereed]
   

  A male infant had the very fragile skin and easily formed bullas by rubbing and scratching from his birth. He was diagnosed with severe recessive dystrophic epidermolysis bullosa (RDEB) due to the lack of type VII collagen by performing an immunofluorescence mapping method from a skin biopsy specimen of the patient's bulla. We analyzed the skin microbiome using next-generation sequencer. The species from the patient's skin revealed the dominance of Staphylococcus aureus (S. aureus) similar to the reports from Austria and Chile severe RDEB patients, and these results are same as the pattern isolated from the skin of atopic dermatitis (AD) patients with flares. The interaction of microbiome and skin microenvironment may be similar between RDEB and AD worldwide.
• Type XVII collagen interacts with the aPKC-PAR complex and maintains epidermal cell polarity.

  Mika Watanabe, Hideyuki Kosumi, Shin-Ichi Osada, Shota Takashima, Yunan Wang, Wataru Nishie, Tsukasa Oikawa, Tomonori Hirose, Hiroshi Shimizu, Ken Natsuga

  Experimental dermatology 30 (1) 62 - 67 2021/01 [Refereed]
   

  Type XVII collagen (COL17) is a transmembrane protein expressed in the basal epidermis. COL17 serves as a niche for epidermal stem cells, and although its reduction has been implicated in altering cell polarity and ageing of the epidermis, it is unknown how COL17 affects epidermal cell polarity. Here, we uncovered COL17 as a binding partner of the aPKC-PAR complex, which is a key regulating factor of cell polarity. Immunoprecipitation-immunoblot assay and protein-protein binding assay revealed that COL17 interacts with aPKC and PAR3. COL17 deficiency or epidermis-specific aPKCλ deletion destabilized PAR3 distribution in the epidermis, while aPKCζ knockout did not. Asymmetrical cell division was pronounced in COL17-null neonatal paw epidermis. These results show that COL17 is pivotal for maintaining epidermal cell polarity. Our study highlights the previously unrecognized role of COL17 in the basal keratinocytes.
• Calcinosis cutis in self-healing dominant dystrophic epidermolysis bullosa.

  Shota Takashima, Yasuyuki Fujita, Satoru Shinkuma, Satoko Shimizu, Tomoka Hasegawa, Norio Amizuka, Hiroshi Shimizu, Ken Natsuga

  The Journal of dermatology 47 (12) e457-e458  2020/12 [Refereed]
  
• Contribution of GATA6 to homeostasis of the human upper pilosebaceous unit and acne pathogenesis.

  Bénédicte Oulès, Christina Philippeos, Joe Segal, Matthieu Tihy, Matteo Vietri Rudan, Ana-Maria Cujba, Philippe A Grange, Sven Quist, Ken Natsuga, Lydia Deschamps, Nicolas Dupin, Giacomo Donati, Fiona M Watt

  Nature communications 11 (1) 5067 - 5067 2020/10/20 [Refereed]
   

  Although acne is the most common human inflammatory skin disease, its pathogenic mechanisms remain incompletely understood. Here we show that GATA6, which is expressed in the upper pilosebaceous unit of normal human skin, is down-regulated in acne. GATA6 controls keratinocyte proliferation and differentiation to prevent hyperkeratinisation of the infundibulum, which is the primary pathological event in acne. When overexpressed in immortalised human sebocytes, GATA6 triggers a junctional zone and sebaceous differentiation program whilst limiting lipid production and cell proliferation. It modulates the immunological repertoire of sebocytes, notably by upregulating PD-L1 and IL10. GATA6 expression contributes to the therapeutic effect of retinoic acid, the main treatment for acne. In a human sebaceous organoid model GATA6-mediated down-regulation of the infundibular differentiation program is mediated by induction of TGFβ signalling. We conclude that GATA6 is involved in regulation of the upper pilosebaceous unit and may be an actionable target in the treatment of acne.
• Multidisciplinary care of epidermolysis bullosa during the COVID-19 pandemic-Consensus: Recommendations by an international panel of experts.

  Dedee F Murrell, Anne W Lucky, Julio C Salas-Alanis, David T Woodley, Francis Palisson, Ken Natsuga, Milos Nikolic, Mae Ramirez-Quizon, Amy S Paller, Irene Lara-Corrales, Mohammadreza Amir Barzegar, Eli Sprecher, Cristina Has, Martin Laimer, Anna L Bruckner, Asli Bilgic, Arti Nanda, Diana Purvis, Alain Hovnanian, Slobodna Murat-Sušić, Johannes Bauer, Johannes S Kern, Christine Bodemer, Linda K Martin, Jemima Mellerio, Cezary Kowaleski, Susan J Robertson, Leena Bruckner-Tuderman, Elena Pope, M Peter Marinkovich, Jean Y Tang, John Su, Jouni Uitto, Lawrence F Eichenfield, Joyce Teng, Mark Jean Aan Koh, Sang Eun Lee, Phuong Khuu, Heather I Rishel, Mette Sommerlund, Karen Wiss, Chao-Kai Hsu, Tor Wo Chiu, Anna E Martinez

  Journal of the American Academy of Dermatology 83 (4) 1222 - 1224 2020/10 [Refereed]
  
• Plectin Missense Mutation p.Leu319Pro in the Pathogenesis of Autosomal Recessive Epidermolysis Bullosa Simplex.

  Wei-Ting Tu, Peng-Chieh Chen, Ping-Chen Hou, Hsin-Yu Huang, Jing-Yu Wang, Sheau-Chiou Chao, Julia Yu-Yun Lee, John A McGrath, Ken Natsuga, Chao-Kai Hsu

  Acta dermato-venereologica 100 (15) adv00242  2020/08/18 [Refereed]
   

  is missing (Short communication).
• Linear IgA/IgG bullous dermatosis with autoantibodies directing the native and processed forms of BP180.

  E Inamura, W Nishie, Y Yamaguchi, Y Fujimura, H Ujiie, K Natsuga, H Shimizu

  The British journal of dermatology 182 (4) 1061 - 1062 2020/04 [Refereed][Not invited]
  
• Speckled lentiginous nevus in a patient with Hermansky-Pudlak syndrome type 1.

  Shoko Mai, Wataru Nishie, Yosuke Mai, Ken Natsuga, Toshifumi Nomura, Shotaro Suzuki, Yuta Araki, Tamio Suzuki, Hiroshi Shimizu

  The Journal of dermatology 47 (1) e20-e21  2020/01 [Refereed][Not invited]
  
• Life before and beyond blistering: The role of collagen XVII in epidermal physiology.

  Ken Natsuga, Mika Watanabe, Wataru Nishie, Hiroshi Shimizu

  Experimental dermatology 28 (10) 1135 - 1141 0906-6705 2019/10 [Refereed][Invited]
   

  Type XVII collagen (COL17) is a transmembranous protein that is mainly expressed in the epidermal basal keratinocytes. Epidermal-dermal attachment requires COL17 expression at the hemidesmosomes of the epidermal basement membrane zone because congenital COL17 deficiency leads to junctional epidermolysis bullosa and acquired autoimmunity to COL17 induces bullous pemphigoid. Recently, in addition to facilitating epidermal-dermal attachment, COL17 has been reported to serve as a niche for hair follicle stem cells, to regulate proliferation in the interfollicular epidermis and to be present along the non-hemidesmosomal plasma membrane of epidermal basal keratinocytes. This review focuses on the physiological properties of COL17 in the epidermis, its role in maintaining stem cells and its association with signalling pathways. We propose possible solutions to unanswered questions in this field.
• Efficient Gene Reframing Therapy for Recessive Dystrophic Epidermolysis Bullosa with CRISPR/Cas9.

  Shota Takashima, Satoru Shinkuma, Yasuyuki Fujita, Toshifumi Nomura, Hideyuki Ujiie, Ken Natsuga, Hiroaki Iwata, Hideki Nakamura, Artem Vorobyev, Riichiro Abe, Hiroshi Shimizu

  The Journal of investigative dermatology 139 (8) 1711 - 1721 0022-202X 2019/08 [Refereed][Not invited]
   

  The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system induces site-specific double-strand breaks, which stimulate cellular DNA repair through either the homologous recombination or non-homologous end-joining pathways. The non-homologous end-joining pathway, which is activated more frequently than homologous recombination, is prone to introducing small insertions and/or deletions at the double-strand break site, leading to changes in the reading frame. We hypothesized that the non-homologous end-joining pathway is applicable to genetic diseases caused by a frameshift mutation through restoration of the reading frame. Recessive dystrophic epidermolysis bullosa is a hereditary skin disorder caused by mutations in COL7A1. In this study, we applied gene reframing therapy to a recurrent frameshift mutation, c.5819delC, in COL7A1, which results in a premature termination codon. CRISPR/Cas9 targeting this specific mutation site was delivered to recessive dystrophic epidermolysis bullosa patient fibroblasts. After genotyping a large collection of gene-edited fibroblast clones, we identified a significant number (17/50) of clones in which the frameshift in COL7A1 was restored. The reframed COL7 was functional, as shown by triple-helix formation assay in vitro, and was correctly distributed in the basement membrane zone in mice. Our data suggest that mutation site-specific non-homologous end-joining might be a highly efficient gene therapy for inherited disorders caused by frameshift mutations.
• Mutant Lef1 controls Gata6 in sebaceous gland development and cancer.

  Bénédicte Oulès, Emanuel Rognoni, Esther Hoste, Georgina Goss, Ryan Fiehler, Ken Natsuga, Sven Quist, Remco Mentink, Giacomo Donati, Fiona M Watt

  The EMBO journal 38 (9) 0261-4189 2019/05/02 [Refereed][Not invited]
   

  Mutations in Lef1 occur in human and mouse sebaceous gland (SG) tumors, but their contribution to carcinogenesis remains unclear. Since Gata6 controls lineage identity in SG, we investigated the link between these two transcription factors. Here, we show that Gata6 is a β-catenin-independent transcriptional target of mutant Lef1. During epidermal development, Gata6 is expressed in a subset of Sox9-positive Lef1-negative hair follicle progenitors that give rise to the upper SG Overexpression of Gata6 by in utero lentiviral injection is sufficient to induce ectopic sebaceous gland elements. In mice overexpressing mutant Lef1, Gata6 ablation increases the total number of skin tumors yet decreases the proportion of SG tumors. The increased tumor burden correlates with impaired DNA mismatch repair and decreased expression of Mlh1 and Msh2 genes, defects frequently observed in human sebaceous neoplasia. Gata6 specifically marks human SG tumors and also defines tumors with elements of sebaceous differentiation, including a subset of basal cell carcinomas. Our findings reveal that Gata6 controls sebaceous gland development and cancer.
• Multiple cutaneous reticulohistiocytomas after haematopoietic cell transplantation: contribution of donor- and host-derived cells.

  H Kosumi, K Natsuga, M Watanabe, K Okada, H Shimizu

  The British journal of dermatology 180 (3) 680 - 681 2019/03 [Refereed]
  
• Image Gallery: Multiple localized lipoatrophy in recessive dystrophic epidermolysis bullosa.

  T Nohara, Y Fujita, S Takashima, K Natsuga, H Shimizu

  The British journal of dermatology 180 (3) e64  2019/03 [Refereed]
  
• Fc-binding proteins enhance autoantibody-induced BP180 depletion in pemphigoid.

  Hiroaki Iwata, Mayumi Kamaguchi, Hideyuki Ujiie, Inkin Ujiie, Ken Natsuga, Wataru Nishie, Hiroshi Shimizu

  The Journal of pathology 247 (3) 371 - 380 0022-3417 2019/03 [Refereed][Not invited]
   

  Immunoglobulins (Igs) consist of two antigen-binding regions (Fab) and one constant region (Fc). Protein A and protein G are bacterial proteins used for the purification of IgG by virtue of their high affinities for the Fc fragment. Rheumatoid factors are autoantibodies against IgG Fc fragments, which are present in the body under physiological conditions. Little is known about the influence of Fc-binding proteins on the pathogenicity of antibody-induced autoimmune diseases. Pemphigoid diseases are a group of autoimmune subepidermal blistering disorders that includes bullous pemphigoid and mucous membrane pemphigoid. IgGs targeting the non-collagenous NC16A domain of the 180-kDa bullous pemphigoid antigen (BP180) are known to induce skin fragility in mice and the depletion of BP180 in keratinocytes. In this study, mAb against NC16A in combination with Fc-binding proteins was found to enhance BP180 depletion. Although mAb against the C-terminus of BP180 does not show pathogenicity in vivo or in vitro, mAb treatment with Fc-binding proteins clearly induced skin fragility in mice and BP180 depletion in keratinocytes. Anti-BP180 mAbs and Fc-binding proteins were colocalized in the cytoplasm and at the basement membrane zone. Cell adhesion strengths were decreased in parallel with BP180 amounts. Clinically, bullous pemphigoid patients had higher rheumatoid factor titers than controls. Anti-BP180 mAb in combination with high-titer rheumatoid factor serum was found to enhance BP180 depletion. Furthermore, saliva from mucous membrane pemphigoid patients contained larger quantities of bacteria and Fc-binding proteins than controls. Our results suggest that Fc-binding proteins (rheumatoid factor or protein G) may enhance the pathogenicity of autoantibodies in pemphigoid diseases. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
• A Nodular Lesion of the Foot Detected by 18F-FDG PET/CT in Mycosis Fungoides: A Plantar Wart.

  Yasuyuki Fujita, Ken Natsuga, Osamu Manabe, Kenji Hirata, Hiroshi Shimizu

  Clinical nuclear medicine 44 (3) 244 - 245 0363-9762 2019/03 [Refereed][Not invited]
   

  A 34-year-old Japanese woman presented with widespread scaly erythema that had enlarged over 2 years. A skin biopsy revealed the diagnosis of mycosis fungoides (patch stage, T1b N0 M0 B0), a most frequent cutaneous T-cell lymphoma. F-FDG PET/CT scan unexpectedly showed intense uptake on the left sole, which suggested a tumorous mycosis fungoides lesion (SUVmax = 6.2). Careful examination revealed the mass to be a typical plantar wart of 2 cm in diameter that the patient had not recognized. With repeated cryotherapy, the wart disappeared in 6 months, and follow-up F-FDG PET/CT showed no abnormal uptake on the left sole.
• Congenital nevi with hypomelanosis and fine scales.

  Ken Natsuga, Naoki Oiso, Ichiro Kurokawa, Airo Tsubura, Hideki Nakamura, Yuka Maya, Wataru Nishie, Akira Kawada, Hiroshi Shimizu

  European journal of dermatology : EJD 29 (1) 45 - 48 1167-1122 2019/02/01 [Refereed][Not invited]
   

  BACKGROUND: Nevus is a hamartoma or malformation of one or more skin components, resulting in aberrant differentiation of the cell lineage(s) mostly during developmental stages. Although multiple lineages may be involved in a nevus, the combination of melanocyte and keratinocyte abnormalities has been rarely discussed. OBJECTIVES: To present two cases of congenital nevi with hypomelanosis and superficial fine scales. MATERIALS & METHODS: Skin specimens of the patients were analysed by immunohistochemistry and electron microscopy. RESULTS: Morphological and immunohistochemical studies indicated aberrant epidermal differentiation in the lesional skin specimens. Electron microscopy showed defective melanosome maturation in the melanocytes of the nevi samples. CONCLUSION: These results demonstrate that both epidermal and melanocytic lineages can concomitantly contribute to the formation of a nevus lesion.
• Immune Reaction to Type XVII Collagen Induces Intramolecular and Intermolecular Epitope Spreading in Experimental Bullous Pemphigoid Models.

  Hideyuki Ujiie, Norihiro Yoshimoto, Ken Natsuga, Ken Muramatsu, Hiroaki Iwata, Wataru Nishie, Hiroshi Shimizu

  Frontiers in immunology 10 1410 - 1410 2019 [Refereed][Not invited]
   

  Bullous pemphigoid (BP), the most common autoimmune blistering disease, is induced by autoantibodies to type XVII collagen (COL17). Previous studies demonstrated that COL17 harbors several epitopes targeted by autoreactive T and B cells and that the target epitopes change sequentially during the disease course. To elucidate the details of the humoral immune response to COL17, we used an active BP mouse model in which BP is induced by the adoptive transfer of spleen cells from wild-type mice immunized with human COL17-expressing skin grafting to immunodeficient COL17-humanized (Rag-2-/-, mouse Col17-/-, human COL17+) mice. By immunoblot analysis, antibodies to the NC16A domain and other extracellular domains (ECDs) of COL17 were detected earlier than antibodies to intracellular domains (ICDs) in the active BP model. Time course analysis by enzyme-linked immunosorbent assay demonstrated a delayed peak of antibodies to ICD epitopes in active BP model. The blockade of CD40-CD40 ligand interaction soon after the adoptive transfer suppressed the production of antibodies to the non-collagenous 16A (NC16A) domain but not to an ICD epitope, suggesting the sequential activation from T and B cells against the ECD epitopes including the NC16A domain to those against ICD epitopes in vivo. Both wild-type mice immunized with a fragment of the NC16A domain and the recipients of those spleen cells produced IgG antibodies to ICD and ECD epitopes, showing intramolecular epitope spreading from the NC16A domain to other epitopes of COL17. Furthermore, we found that a portion of the active BP model mice show intermolecular epitope spreading from human COL17 to murine BP230. The appearance of antibodies to ICD epitopes of COL17 or of antibodies to murine BP230 did not correlate with the skin changes in the mice, suggesting that those antibodies have low pathogenicity. These results suggest that the immune response to the ECD epitopes of COL17, especially to the NC16A domain, triggers intramolecular, and intermolecular epitope spreading to ICD epitopes of COL17 and to murine BP230. These novel findings provide insight into the mechanism of epitope spreading in organ-specific, antibody-mediated autoimmune disorders.
• The direct binding of collagen XVII and collagen IV is disrupted by pemphigoid autoantibodies.

  Mayumi Kamaguchi, Hiroaki Iwata, Wataru Nishie, Ellen Toyonaga, Hideyuki Ujiie, Ken Natsuga, Yoshimasa Kitagawa, Hiroshi Shimizu

  Laboratory investigation; a journal of technical methods and pathology 99 (1) 48 - 57 0023-6837 2019/01 [Refereed][Not invited]
   

  The basement membrane zone (BMZ) is framed by hemidesmosomes and extracellular matrix (ECM) including collagen IV (COL4). Hemidesmosomes are multiprotein complexes that include collagen XVII (COL17). BMZ proteins can be targeted in autoimmune subepidermal blistering diseases, e.g., pemphigoid targeting COL17. The blistering mechanisms in pemphigoid have not been fully elucidated, especially in mucous membrane pemphigoid (MMP), which mainly affects the mucosa. In this study, we showed that oral lesions in pemphigoid may be attributed to the inhibition of protein-protein interactions by autoantibodies. Using immunoprecipitation, we revealed that COL17 directly binds to COL4 in normal human keratinocytes and normal human oral keratinocytes. In particular, the C-terminus of COL17 is binding site to COL4 in oral keratinocytes. The precise COL4-binding region on COL17 was determined by protein-protein binding assay to be from amino acid Gly1175 to Asp1340 on the C-terminus. MMP-IgG or mAb recognizing the C-terminus hindered the interaction of COL17 with COL4 in oral keratinocytes. Furthermore, keratinocyte adhesion strength to COL4-coated plates was significantly reduced by the treatment of mAb against the C-terminus. In addition, the inflammatory infiltrates around perilesions were significantly less in MMP compared to BP. These results indicate that pemphigoid IgG targeting the C-terminus plays a pathogenic role in blister formation in the oral mucosa to inhibit protein interactions with less inflammation.
• Bullous Pemphigoid IgG Induces Cell Dysfunction and Enhances the Motility of Epidermal Keratinocytes via Rac1/Proteasome Activation.

  Duerna Tie, Xia Da, Ken Natsuga, Nanako Yamada, Osamu Yamamoto, Eishin Morita

  Frontiers in immunology 10 200 - 200 2019 [Refereed][Not invited]
   

  Bullous pemphigoid (BP) is an autoimmune disease characterized by the formation of blisters, in which autoantibodies mainly target type XVII collagen (ColXVII) expressed in basal keratinocytes. BP IgG is known to induce the internalization of ColXVII from the plasma membrane of keratinocytes through macropinocytosis. However, the cellular dynamics following ColXVII internalization have not been completely elucidated. BP IgG exerts a precise effect on cultured keratinocytes, and the morphological/functional changes in BP IgG-stimulated cells lead to the subepidermal blistering associated with BP pathogenesis. Based on the electron microscopy examination, BP IgG-stimulated cells exhibit alterations in the cell membrane structure and the accumulation of intracellular vesicles. These morphological changes in the BP IgG-stimulated cells are accompanied by dysfunctional mitochondria, increased production of reactive oxygen species, increased motility, and detachment. BP IgG triggers the cascade leading to metabolic impairments and stimulates cell migration in the treated keratinocytes. These cellular alterations are reversed by pharmacological inhibitors of Rac1 or the proteasome pathway, suggesting that Rac1 and proteasome activation are involved in the effects of BP IgG on cultured keratinocytes. Our study highlights the role of keratinocyte kinetics in the direct functions of IgG in patients with BP.
• Interplay between epidermal stem cell dynamics and dermal deformation

  Yasuaki Kobayashi, Yusuke Yasugahira, Hiroyuki Kitahata, Mika Watanabe, Ken Natsuga, Masaharu Nagayama

  npj Computational Materials 4 2018/12 [Refereed][Not invited]
   

  Tissue growth is a driving force of morphological changes in living systems. Whereas the buckling instability is known to play a crutial role for initiating spatial pattern formations in such growing systems, little is known about the rationale for succeeding morphological changes beyond this instability. In mammalian skin, the dermis has many protrusions toward the epidermis, and the epidermal stem cells are typically found on the tips of these protrusions. Although the initial instability may well be explained by the buckling involving the dermis and the basal layer, which contains proliferative cells, it does not dictate the direction of these protrusions, nor the spatial patterning of epidermal stem cells. Here we introduce a particle-based model of self-replicating cells on a deformable substrate composed of the dermis and the basement membrane, and investigate the relationship between dermal deformation and epidermal stem cell pattering on it. We show that our model reproduces the formation of dermal protrusions directing from the dermis to the epidermis, and preferential epidermal stem cell distributions on the tips of the dermal protrusions, which the basic buckling mechanism fails to explain. We argue that cell-type-dependent adhesion strengths of the cells to the basement membrane are crucial factors influencing these patterns.
• Regulatory T-cell dysfunction induces autoantibodies to bullous pemphigoid antigens in mice and human subjects.

  Ken Muramatsu, Hideyuki Ujiie, Ichiro Kobayashi, Wataru Nishie, Kentaro Izumi, Takamasa Ito, Norihiro Yoshimoto, Ken Natsuga, Hiroaki Iwata, Hiroshi Shimizu

  The Journal of allergy and clinical immunology 142 (6) 1818 - 1830 0091-6749 2018/12 [Refereed][Not invited]
   

  BACKGROUND: Regulatory T (Treg) cells play a crucial role in peripheral immune tolerance in multiple organs, including the skin. Thus far, the effect of peripheral immune tolerance failure on autoantibody-related autoimmune reactions to the skin is unclear. OBJECTIVE: We sought to elucidate the target autoantigens in the skin under the condition of Treg cell dysfunction caused by forkhead box P3 (Foxp3) gene mutations in scurfy mice and patients with immunodysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome. METHODS: Sera and skin from scurfy mice and sera from patients with IPEX syndrome were analyzed to detect target autoantigens by using immunofluorescence studies, ELISAs, and immunoblotting. The pathogenicity of scurfy IgG was examined by using a passive transfer experiment. CD4+ T cells from scurfy mice were transferred to immunodeficient mice to examine their pathogenicity. Signal transducer and activator of transcription 6 (Stat6)-/- scurfy mice were analyzed to further clarify the molecular pathway of autoantibody production. Follicular helper T-cell counts are measured in Stat6-/- scurfy mice and scurfy mice. RESULTS: Scurfy mice spontaneously generated IgG autoantibodies to the dermal-epidermal junction, which had been class-switched from IgM within 12 days after birth. The target autoantigens were murine BP230 and type XVII collagen (COL17). The scurfy polyclonal autoantibodies did not induce skin fragility in neonatal mice. Autoantibody production was induced by CD4+ T cells from scurfy mice and was ameliorated by Stat6 gene knockout in association with a decrease of follicular helper T cells. We also identified autoantibodies to COL17 and BP230 in patients with IPEX syndrome and found an association between production of autoantibodies to COL17 and an eczematous skin phenotype. CONCLUSIONS: Dysregulation of Treg cells generates autoantibodies to COL17 and BP230 in vivo.
• Novel COL7A1 mutation in a family with bullous dermolysis of the newborn: Phenotypic variability associated with a COL7A1 mutation within the same family.

  Shota Takashima, Satoru Shinkuma, Yasuyuki Fujita, Ken Natsuga, Toshifumi Nomura, Tokimasa Hida, Shuku Ishikawa, Hideki Nakamura, Riichiro Abe, Hiroshi Shimizu

  The Journal of dermatology 45 (9) e260-e261 - e261 0385-2407 2018/09 [Refereed][Not invited]
  
• Remission of Cutaneous Involvement in Splenic Marginal Zone Lymphoma after Splenectomy.

  Shoko Mai, Ken Natsuga, Souichi Shiratori, Yoshimasa Takahashi, Hiroshi Shimizu

  Acta dermato-venereologica 98 (8) 801 - 802 0001-5555 2018/08/29 [Refereed][Not invited]
  
• High Expression of Collagen XVII Compensates for its Depletion Induced by Pemphigoid IgG in the Oral Mucosa.

  Mayumi Kamaguchi, Hiroaki Iwata, Hideyuki Ujiie, Ken Natsuga, Wataru Nishie, Yoshimasa Kitagawa, Hiroshi Shimizu

  The Journal of investigative dermatology 138 (8) 1707 - 1715 0022-202X 2018/08 [Refereed][Not invited]
   

  The basement membrane zone consists of multiple components, including collagen XVII (COL17), which is the target of bullous pemphigoid. To our knowledge, no research has addressed the differences in basement membrane zone components between the skin and oral mucosa; therefore, we investigated the basement membrane zone proteins, with a focus on COL17. The mRNA and protein expression levels of COL17 were significantly higher in oral keratinocytes than in skin keratinocytes. Hemidesmosomal COL17 expression was markedly higher in oral keratinocytes than in skin keratinocytes, and its level was associated with adhesion strength. Oral keratinocytes adhered to the extracellular matrix more tightly than did skin keratinocytes in vitro. Based on these results, we attempt to explain the clinical diversity of bullous pemphigoid. COL17 depletion was more prominent in skin keratinocytes than in oral keratinocytes after treatment with COL17-NC16A mAbs, which have in vivo pathogenicity. COL17 C-terminus mAbs, which are not pathogenic, facilitated COL17 depletion in combination treatment with COL17-NC16A mAbs in both types of keratinocytes. In summary, the greater amount of COL17 in oral keratinocytes than in skin keratinocytes is associated with the higher strength of oral keratinocyte hemidesmosomal adhesion at the basement membrane zone. Our results may explain why bullous pemphigoid blistering tends to be more prevalent in the skin than in the oral mucosa.
• Acute vascular reaction due to lipo-prostaglandin E1.

  Y Fujimura, M Watanabe, K Natsuga, H Shimizu

  Journal of the European Academy of Dermatology and Venereology : JEADV 32 (7) e273-e274  2018/07 [Refereed]
  
• Sweet's Syndrome Mimicking Anti-Neutrophil Cytoplasmic Antibodies-Associated Vasculitis.

  Hideyuki Kosumi, Mika Watanabe, Ken Natsuga, Toshinari Miyauchi, Chihiro Shiiya, Hideyuki Ujiie, Hiroshi Shimizu

  The American journal of medicine 131 (6) e241-e242 - e242 0002-9343 2018/06 [Refereed][Not invited]
  
• Intravenous IgG Reduces Pathogenic Autoantibodies, Serum IL-6 Levels, and Disease Severity in Experimental Bullous Pemphigoid Models.

  Tetsumasa Sasaoka, Hideyuki Ujiie, Wataru Nishie, Hiroaki Iwata, Makoto Ishikawa, Hiroshi Higashino, Ken Natsuga, Satoru Shinkuma, Hiroshi Shimizu

  The Journal of investigative dermatology 138 (6) 1260 - 1267 0022-202X 2018/06 [Refereed][Not invited]
   

  Bullous pemphigoid (BP) is an autoimmune blistering disease characterized by autoantibodies to COL17. Currently, systemic corticosteroids are used as first-line treatments for BP; alternatively, intravenous administration of high-dose IgG (IVIG) has been shown to be effective for patients with steroid-resistant BP in clinical practice. However, the effect of IVIG on BP has not fully been investigated. To examine the effects and mechanisms of action of IVIG against BP, we performed IVIG experiments using two experimental BP mouse models. One is a passive-transfer BP model that reproduces subepidermal separation in neonatal mice by the passive transfer of IgGs against COL17, such as polyclonal or monoclonal mouse IgG or IgG from BP patients. The other is an active BP model that continuously develops a disease phenotype in adult mice. IVIG decreased pathogenic IgG and the disease scores in both models. Injected IVIG distributed throughout the dermis and the intercellular space of the lower epidermis. Notably, IVIG inhibited the increase of IL-6 in both models, possibly by suppressing the production of IL-6 by keratinocytes. These results suggest that the inhibitory effects of IVIG on BP are associated with the reduction of pathogenic IgG and the modulation of cytokine production.
• Epidermal aspects of type VII collagen: Implications for dystrophic epidermolysis bullosa and epidermolysis bullosa acquisita.

  Mika Watanabe, Ken Natsuga, Satoru Shinkuma, Hiroshi Shimizu

  The Journal of dermatology 45 (5) 515 - 521 0385-2407 2018/05 [Refereed][Not invited]
   

  Type VII collagen (COL7), a major component of anchoring fibrils in the epidermal basement membrane zone, has been characterized as a defective protein in dystrophic epidermolysis bullosa and as an autoantigen in epidermolysis bullosa acquisita. Although COL7 is produced and secreted by both epidermal keratinocytes and dermal fibroblasts, the role of COL7 with regard to the epidermis is rarely discussed. This review focuses on COL7 physiology and pathology as it pertains to epidermal keratinocytes. We summarize the current knowledge of COL7 production and trafficking, its involvement in keratinocyte dynamics, and epidermal carcinogenesis in COL7 deficiency and propose possible solutions to unsolved issues in this field.
• Portable negative-pressure wound therapy for pyoderma gangrenosum: Report of two cases.

  Yasuyuki Yamaguchi, Teruki Yanagi, Kazumasa Sato, Norihiro Yoshimoto, Yu Hirata, Inkin Ujiie, Machiko Nishimura, Ken Natsuga, Chihiro Shiiya, Ichiro Tsukinaga, Hiroshi Shimizu

  The Journal of dermatology 45 (4) 483 - 486 0385-2407 2018/04 [Refereed][Not invited]
   

  Pyoderma gangrenosum is a chronic non-infectious neutrophilic dermatosis that causes undermining ulcers. Topical therapies for the deep ulcers of pyoderma gangrenosum have not been established. To investigate whether negative-pressure wound therapy is effective for a pyoderma gangrenosum ulcer, we used the PICO single use negative-pressure wound therapy system (Smith & Nephew, London, UK) for two pyoderma gangrenosum patients. In these cases, the ulcers decreased in size and necrolytic tissue was removed notably. Moreover, there were no secondary infections nor was there Koebner phenomena. Our cases suggest that portable negative-pressure wound therapy can be a treatment option for deep, intractable ulcers caused by pyoderma gangrenosum. Because portable negative-pressure wound therapy devices afford increased mobility to patients, they can give the patient a better quality of life than standard negative-pressure wound therapy systems do.
• Oral mucosa is a useful substrate for detecting autoantibodies of mucous membrane pemphigoid

  M. Kamaguchi, H. Iwata, H. Ujiie, K. Izumi, K. Natsuga, W. Nishie, T. Asaka, Y. Kitagawa, H. Shimizu

  British Journal of Dermatology 178 (2) e119 - e121 1365-2133 2018/02/01 [Refereed][Not invited]
  
• Extensive Eruptive Syringoma After Liver Transplantation.

  Takuya Maeda, Ken Natsuga, Wataru Nishie, Kenichiro Yamashita, Hiroshi Shimizu

  Acta dermato-venereologica 98 (1) 119 - 120 0001-5555 2018/01/12 [Refereed][Not invited]
  
• Appearance of antidesmocollin 1 autoantibodies leading to a vegetative lesion in a patient with pemphigus vulgaris

  Y. Yamaguchi, S. Shinkuma, N. Ishii, S. Takashima, K. Natsuga, H. Ujiie, H. Iwata, T. Nomura, Y. Fujita, A. Hamasaka, K. Hamasaka, T. Hashimoto, H. Shimizu

  British Journal of Dermatology 178 (1) 294 - 295 1365-2133 2018/01/01 [Refereed][Not invited]
  
• C-Terminal Processing of Collagen XVII Induces Neoepitopes for Linear IgA Dermatosis Autoantibodies

  Ellen Toyonaga, Wataru Nishie, Kentaro Izumi, Ken Natsuga, Hideyuki Ujiie, Hiroaki Iwata, Jun Yamagami, Yoshiaki Hirako, Daisuke Sawamura, Wataru Fujimoto, Hiroshi Shimizu

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 137 (12) 2552 - 2559 0022-202X 2017/12 [Refereed][Not invited]
   

  Transmembrane collagen XVII (COL17) is a hemidesmosomal component of basal keratinocytes that can be targeted by autoantibodies in autoimmune blistering disorders, including linear IgA dermatosis (LAD). COL17 can be physiologically cleaved within the juxtamembranous extracellular NC16A domain, and LAD autoantibodies preferentially react with the processed ectodomains, indicating that the processing induces neoepitopes. However, the details of how neoepitopes develop have not been elucidated. In this study, we show that C-terminal processing of COL17 also plays a role in inducing neoepitopes for LAD autoantibodies. First, the mAb hC17-ect15 targeting the 15th collagenous domain of COL17 was produced, which showed characteristics similar to LAD autoantibodies. The mAbs preferentially reacted with C-terminally deleted (up to 682 amino acids) recombinant COL17, suggesting that C-terminal processing shows neoepitopes on the 15th collagenous domain. The LAD autoantibodies also react with C-terminal deleted COL17. Therefore, neoepitopes for LAD autoantibodies also develop after C-terminal processing. Finally, the passive transfer of the mAb hC17-ect15 into human COL17-expressing transgenic mice failed to induce blistering disease, suggesting that neoepitope-targeting antibodies are not always pathogenic. In summary, this study shows that C-terminal processing induces dynamic structural changes and neoepitopes for LAD autoantibodies on COL17.
• Loss of interaction between plectin and type XVII collagen results in epidermolysis bullosa simplex

  Ken Natsuga, Wataru Nishie, Machiko Nishimura, Satoru Shinkuma, Mika Watanabe, Kentaro Izumi, Hideki Nakamura, Yoshiaki Hirako, Hiroshi Shimizu

  HUMAN MUTATION 38 (12) 1666 - 1670 1059-7794 2017/12 [Refereed][Not invited]
   

  Plectin is a linker protein that interacts with intermediate filaments and 4 integrin in hemidesmosomes of the epidermal basement membrane zone (BMZ). Type XVII collagen (COL17) has been suggested as another candidate plectin binding partner in hemidesmosomes. Here, we demonstrate that plectin-COL17 binding helps to maintain epidermal BMZ organization. We identified an epidermolysis bullosa (EB) simplex patient as having markedly diminished expression of plectin and COL17 in skin. The patient is compound heterozygous for sequence variants in the plectin gene (PLEC); one is a truncation and the other is a small in-frame deletion sequence variant. The in-frame deletion is located in the putative COL17-binding domain of plectin and abolishes the plectin-COL17 interaction in vitro. These results imply that disrupted interaction between plectin and COL17 is involved in the development of EB. Our study suggests that protein-protein binding defects may underlie EB in patients with unidentified disease-causing sequence variants.
• Autoantibodies of non-inflammatory bullous pemphigoid hardly deplete type XVII collagen of keratinocytes

  Keisuke Imafuku, Hiroaki Iwata, Mayumi Kamaguchi, Kentaro Izumi, Ken Natsuga, Hideyuki Ujiie, Wataru Nishie, Hiroshi Shimizu

  Experimental Dermatology 26 (12) 1171 - 1174 1600-0625 2017/12/01 [Refereed][Not invited]
   

  Type XVII collagen (COL17) and the non-collagenous 16A (NC16A) domain is regarded as the major pathogenic domains for bullous pemphigoid (BP). Some patients with BP have autoantibodies against parts of COL17 outside the NC16A domain (hereinafter the non-NC16A domain) and show less inflammatory manifestations. There were no significant differences in titres and IgG subclasses between NC16A-BP and non-NC16A-BP as determined by indirect immunofluorescent microscopy. The neutrophil activation capacities determined by ROS release did not differ between NC16A-BP and non-NC16A-BP. However, NC16A-BP IgG depleted COL17 in a dose-dependent manner. Treatment with NC16A-BP IgG, but not with non-NC16A-BP IgG, significantly decreased the adhesion strength. We speculate that the differences in clinical severity between NC16A-BP and non-NC16A-BP relate to the degree of COL17 depletion.
• Cutis verticis gyrata fluctuation with atopic dermatitis disease activity

  Hideyuki Kosumi, Kentaro Izumi, Ken Natsuga, Yasuyuki Yamaguchi, Akira Itami, Hiroshi Shimizu

  Acta Dermato-Venereologica 97 (10) 1245 - 1246 1651-2057 2017/11/01 [Refereed][Not invited]
  
• Detection of mucous membrane pemphigoid autoantibodies by full-length BP180 enzyme-linked immunosorbent assay

  Kentaro Izumi, Wataru Nishie, Yosuke Mai, Hideyuki Ujiie, Hiroaki Iwata, Ken Natsuga, Hiroshi Shimizu

  JOURNAL OF DERMATOLOGICAL SCIENCE 88 (2) 247 - 248 0923-1811 2017/11 [Refereed][Not invited]
  
• A case of recessive dystrophic epidermolysis bullosa with a novel c.6885_6898de114 mutation in the COL7A1 gene

  Satoru Shinkuma, Tae Masunaga, Saori Miyawaki, Shota Takashima, Ken Natsuga, Toshifumi Nomura, Yasuyuki Fujita, Hideki Nakamura, Hiroshi Shimizu

  JOURNAL OF DERMATOLOGICAL SCIENCE 88 (1) 139 - 141 0923-1811 2017/10 [Refereed][Not invited]
  
• Generalized Pustular Psoriasis

  Hideyuki Kosumi, Takamasa Ito, Yasuyuki Fujita, Kentaro Izumi, Yuka Maya, Teruki Yanagi, Ken Natsuga, Hideyuki Ujiie, Satoru Shinkuma, Toshifumi Nomura, Naoya Sadanobu, Hiroshi Shimizu

  JOURNAL OF PEDIATRICS 188 305 - + 0022-3476 2017/09 [Refereed][Not invited]
  
• Type XVII collagen coordinates proliferation in the interfollicular epidermis

  Mika Watanabe, Ken Natsuga, Wataru Nishie, Yasuaki Kobayashi, Giacomo Donati, Shotaro Suzuki, Yu Fujimura, Tadasuke Tsukiyama, Hideyuki Ujiie, Satoru Shinkuma, Hideki Nakamura, Masamoto Murakami, Michitaka Ozaki, Masaharu Nagayama, Fiona M. Watt, Hiroshi Shimizu

  ELIFE 6 2050-084X 2017/07 [Refereed][Not invited]
   

  Type XVII collagen (COL17) is a transmembrane protein located at the epidermal basement membrane zone. COL17 deficiency results in premature hair aging phenotypes and in junctional epidermolysis bullosa. Here, we show that COL17 plays a central role in regulating interfollicular epidermis (IFE) proliferation. Loss of COL17 leads to transient IFE hypertrophy in neonatal mice owing to aberrant Wnt signaling. The replenishment of COL17 in the neonatal epidermis of COL17-null mice reverses the proliferative IFE phenotype and the altered Wnt signaling. Physical aging abolishes membranous COL17 in IFE basal cells because of inactive atypical protein kinase C signaling and also induces epidermal hyperproliferation. The overexpression of human COL17 in aged mouse epidermis suppresses IFE hypertrophy. These findings demonstrate that COL17 governs IFE proliferation of neonatal and aged skin in distinct ways. Our study indicates that COL17 could be an important target of anti-aging strategies in the skin.
• Crusted impetigo-like lesion on the face: a case of IgG/IgA pemphigus

  R. Moriuchi, T. Ito, K. Kikuchi, N. Nakashita, R. Muramatsu, K. Natsuga, H. Shimizu, S. Shimizu

  JOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY AND VENEREOLOGY 31 (6) E289 - E290 0926-9959 2017/06 [Refereed][Not invited]
   

  Moriuchi R, Ito T, Kikuchi K, Nakashita N, Muramatsu R, Natsuga K, Shimizu H, Shimizu S, Journal of the European Academy of Dermatology and Venereology : JEADV, 2016, 2016
• Psoriasiform mycosis fungoides mas-querading as tumourous plaques

  Yasuyuki Yamaguchi, Yasuyuki Fujita, Yu Hirata, Machiko Nishimura, Satoru Shinkuma, Ken Natsuga, Toshifumi Nomura, Tokimasa Hida, Naoko Kato, Hiroshi Shimizu

  EUROPEAN JOURNAL OF DERMATOLOGY 27 (3) 295 - 296 1167-1122 2017/05 [Refereed][Not invited]
   

  Yamaguchi Y, Fujita Y, Hirata Y, Nishimura M, Shinkuma S, Natsuga K, Nomura T, Hida T, Kato N, Shimizu H, European journal of dermatology : EJD, 2017, 2017
• Early severe pachyonychia congenita subtype PC-K6a with a novel mutation in the KRT6A gene

  F. Cammarata-Scalisi, K. Natsuga, E. Toyonaga, W. Nishie, H. Shimizu, A. Avendano, D. Araque, G. Da Silva, E. Bellacchio, M. Callea

  JOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY AND VENEREOLOGY 31 (2) E94 - E96 0926-9959 2017/02 [Refereed][Not invited]
   

  Cammarata-Scalisi F, Natsuga K, Toyonaga E, Nishie W, Shimizu H, Avendaño A, Araque D, Da Silva G, Bellacchio E, Callea M, Journal of the European Academy of Dermatology and Venereology : JEADV, 2017, vol. 31, no. 2, pp. e94-e96, 2017
• Lichenoid drug eruption caused by clonazepam

  K. Muramatsu, H. Ujiie, K. Natsuga, W. Nishie, H. Shimizu

  JOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY AND VENEREOLOGY 31 (2) E117 - E118 0926-9959 2017/02 [Refereed][Not invited]
   

  Muramatsu K, Ujiie H, Natsuga K, Nishie W, Shimizu H, Journal of the European Academy of Dermatology and Venereology : JEADV, 2017, vol. 31, no. 2, pp. e117-e118, 2017
• Macropinocytosis of type XVII collagen induced by bullous pemphigoid IgG is regulated via protein kinase C

  Hiroaki Iwata, Mayumi Kamaguchi, Hideyuki Ujiie, Machiko Nishimura, Kentaro Izumi, Ken Natsuga, Satoru Shinkuma, Wataru Nishie, Hiroshi Shimizu

  LABORATORY INVESTIGATION 96 (12) 1301 - 1310 0023-6837 2016/12 [Refereed][Not invited]
   

  Macropinocytosis is an endocytic pathway that is involved in the nonselective fluid uptake of extracellular fluid. Bullous pemphigoid (BP) is an autoimmune subepidermal blistering disease associated with autoantibodies to type XVII collagen (COL17), which is a component of hemidesmosome. When keratinocytes are treated with BP-IgG, COL17 internalizes into cells by way of the macropinocytosis. We investigated the mechanism of COL17 macropinocytosis using DJM-1 cells, a cutaneous squamous cell carcinoma cell line. First, non-hemidesmosomal COL17 was preferentially depleted by stimulation with the BP-IgG in the DJM-1 cells. To investigate the signaling involved in COL17-macropinocytosis, the inhibition of small GTPase family members Rac1 and Cdc42 was found to strongly repress COL17 internalization; in addition, the Rho inhibitor also partially blocked that internalization, suggesting these small GTPases are involved in signaling to mediate COL17-macropinocytosis. Western blotting using Phostag-SDS-PAGE demonstrated high levels of COL17 phosphorylation in DJM-1 cells under steady-state condition. Treatment with BP-IgG increased the intracellular calcium level within a minute, and induced the overabundant phosphorylation of COL17. The overabundant phosphorylation of COL17 was suppressed by a protein kinase C (PKC) inhibitor. In addition, PKC inhibitor repressed COL17 endocytosis using cell culture and organ culture systems. Finally, the depletion of COL17 was not observed in the HEK293 cells transfected COL17 without intracellular domain. These results suggest that COL17 internalization induced by BP-IgG may be mediated by a PKC pathway. In summary, BP-IgG initially binds to COL17 distributed on the plasma membrane, and COL17 may be internalized by means of a macropinocytic pathway related to the phosphorylation of the intracellular domain by PKC.
• Generalized acute subcutaneous edema as a rare cutaneous manifestation of severe dermatomyositis

  M. Watanabe, K. Natsuga, K. Arita, R. Abe, H. Shimizu

  JOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY AND VENEREOLOGY 30 (11) E151 - E152 0926-9959 2016/11 [Refereed][Not invited]
   

  Watanabe M, Natsuga K, Arita K, Abe R, Shimizu H, Journal of the European Academy of Dermatology and Venereology : JEADV, 2015
• Two cases of erosive oral lichen planus with autoantibodies to desmoglein 3

  Ken Muramatsu, Wataru Nishie, Ken Natsuga, Yasuyuki Fujita, Hiroaki Iwata, Tamaki Yamada, Emi Yamashita, Takuya Asaka, Hiroshi Shimizu

  JOURNAL OF DERMATOLOGY 43 (11) 1350 - 1353 0385-2407 2016/11 [Refereed][Not invited]
   

  Oral lichen planus (OLP) is a chronic inflammatory disorder of the oral mucosa of unknown etiology. Clinically, the erosive type of OLP (erosive OLP) can show features similar to those of pemphigus vulgaris (PV), an autoimmune blistering disorder in which desmoglein (Dsg) 3 is targeted. In addition to clinical and histopathological findings, immunological studies, including direct immunofluorescence (IF), indirect IF and enzyme-linked immunosorbent assay (ELISA) that detect autoantibodies to Dsg3, are helpful in differentiating erosive OLP from PV. Here, we show two cases of erosive OLP with autoantibodies to Dsg3. Patient 1 was a 68-year-old woman with chronic erosions of the oral mucosa, in which elevated levels of immunoglobulin (Ig)G autoantibodies to Dsg1 and Dsg3 were detected by ELISA. Patient 2 was an 85-year-old woman with white striae with erosions on the lateral sides of the buccal mucosa with elevated levels of IgG autoantibodies to Dsg3 detected by ELISA. Histopathological findings from both cases showed lichenoid dermatitis, and both direct and indirect IF showed no tissue-bound IgG autoantibodies. From these findings, the diagnosis of erosive OLP was made. Immunological assays revealed both cases to have IgG-directing calcium-independent linear epitopes on Dsg3, which are suggestive of non-pathogenic autoantibodies. In addition, autoantibodies to Dsg3 in patient 2 reacted with a prosequence-possessing precursor form of Dsg3 but not with the mature form of the molecule. The present study suggests that erosive OLP may develop anti-Dsg3 autoantibodies, which should be carefully assessed.
• Autoantibody Profile Differentiates between Inflammatory and Noninflammatory Bullous Pemphigoid

  Kentaro Izumi, Wataru Nishie, Yosuke Mai, Mayumi Wada, Ken Natsuga, Hideyuki Ujiie, Hiroaki Iwata, Jun Yamagami, Hiroshi Shimizu

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 136 (11) 2201 - 2210 0022-202X 2016/11 [Refereed][Not invited]
   

  Bullous pemphigoid (BP) is a major autoimmune blistering skin disorder, in which a majority of the autoantibodies (autoAbs) target the juxtamembranous extracellular noncollagenous 16A domain (NC16A) domain of hemidesmosomal collagen XVII. BP-autoAbs may target regions of collagen XVII other than the NC16A domain; however, correlations between epitopes of BP-autoAbs and clinical features have not been fully elucidated. To address correlations between the clinical features and specific epitopes of BP-autoAbs, we evaluated the epitope profiles of BP-autoAbs in 121 patients. A total of 87 patients showed a typical inflammatory phenotype with erythema and autoAbs targeting the anti-NC16A domain, whereas 14 patients showed a distinct noninflammatory phenotype, in which autoAbs specifically targeted the midportion of collagen XVII, but not NC16A. Interestingly, this group clinically showed significantly reduced erythema associated with scant lesional infiltration of eosinophils. Surprisingly, 7 of the 14 cases (50.0%) received dipeptidyl peptidase-IV inhibitors for the treatment of diabetes. Dipeptidyl peptidase-IV inhibitors were used in 3 of 76 (3.9%) typical cases of BP with autoAbs targeting NC16A; thus, dipeptidyl peptidase-IV inhibitors are thought to be involved in the development of atypical noninflammatory BP. This study shows that the autoAb profile differentiates between inflammatory and noninflammatory BP, and that noninflammatory BP may be associated with dipeptidyl peptidase-IV inhibitors.
• Galectin-6 is a novel skin anti-microbial peptide that is modulated by the skin barrier and microbiome

  Ken Natsuga, Fiona M. Watt

  JOURNAL OF DERMATOLOGICAL SCIENCE 84 (1) 97 - 99 0923-1811 2016/10 [Refereed][Not invited]
   

  Natsuga K, Watt FM, Journal of dermatological science, 2016, vol. 84, no. 1, pp. 97-99, 2016
• Stem cell markers of skin appendages in human skin tumours

  K. Natsuga

  BRITISH JOURNAL OF DERMATOLOGY 175 (3) 459 - 459 0007-0963 2016/09 [Not refereed][Invited]
   

  Linked Article: Quist et al. Br J Dermatol 2016; 175:520-530.
• Silicone medical adhesive removers for hyperkeratosis in epidermolysis bullosa

  Yasuyuki Fujita, Satoru Shinkuma, Wakana Matsumura, Ken Natsuga, Hiroo Hata, Toshifumi Nomura, Hiroshi Shimizu

  EUROPEAN JOURNAL OF DERMATOLOGY 26 (5) 501 - 502 1167-1122 2016/09 [Refereed][Not invited]
   

  Fujita Y, Shinkuma S, Matsumura W, Natsuga K, Hata H, Nomura T, Shimizu H, European journal of dermatology : EJD, 2016, vol. 26, no. 5, pp. 501-502, 2016
• mTOR expression correlates with invasiveness and progression of extramammary Paget's disease

  H. Hata, S. Kitamura, Y. Inamura, K. Imafuku, E. Homma, K. Muramatsu, K. Natsuga, R. Abe, H. Shimizu

  JOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY AND VENEREOLOGY 30 (7) 1238 - 1239 0926-9959 2016/07 [Refereed][Not invited]
   

  Hata H, Kitamura S, Inamura Y, Imafuku K, Homma E, Muramatsu K, Natsuga K, Abe R, Shimizu H, Journal of the European Academy of Dermatology and Venereology : JEADV, 2015
• Verrucous hyperplasia associated with neuropathy from schistorrhachis

  K. Imafuku, H. Hata, Y. Inamura, S. Kitamura, K. Natsuga, H. Iwata, H. Shimizu

  JOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY AND VENEREOLOGY 30 (5) 876 - 878 0926-9959 2016/05 [Refereed][Not invited]
   

  Imafuku K, Hata H, Inamura Y, Kitamura S, Natsuga K, Iwata H, Shimizu H, Journal of the European Academy of Dermatology and Venereology : JEADV, 2015
• Epitope-Dependent Pathogenicity of Antibodies Targeting a Major Bullous Pemphigoid Autoantigen Collagen XVII/BP180

  Mayumi Wada, Wataru Nishie, Hideyuki Ujiie, Kentaro Izumi, Hiroaki Iwata, Ken Natsuga, Hideki Nakamura, Yoshimasa Kitagawa, Hiroshi Shimizu

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 136 (5) 938 - 946 0022-202X 2016/05 [Refereed][Not invited]
   

  In bullous pemphigoid, the common autoimmune blistering disorder, IgG autoantibodies target various epitopes on hemidesmosomal transmembrane collagen XVII (COL17)/BP180. Antibodies (Abs) targeting the extracellular noncollagenous 16th A domain of COL17 may be pathogenic; however, the pathogenic roles of Abs targeting non-noncollagenous 16th A regions are poorly understood. In this study using a pathogenic and a nonpathogenic monoclonal antibody (mAb) targeting the noncollagenous 16th A domain (mAb TS39-3) and the C-terminus domain (mAb C17-C1), respectively, we show that endocytosis of immune complexes after binding of Abs to cell surface COL17 is a key phenomenon that induces skin fragility. Passive transfer of IgG1 mouse mAb TS39-3 but not mAb C17-C1 induces dermal-epidermal separation in neonatal human COL17-expressing transgenic mice. Interestingly, mAb C17-C1 strongly binds with the dermal-epidermal junction of the recipient mice skin, suggesting that binding of Abs with COL17 is insufficient to induce skin fragility. In cultured normal human epidermal keratinocytes treated with these mAbs, mAb TS39-3 but not mAb C17-C1 internalizes immune complexes after binding with cell surface COL17 via macropinocytosis, resulting in reduced COL17 expression. This study shows that pathogenicity of Abs targeting COL17 is epitope dependent, which is associated with macropinocytosis-mediated endocytosis of immune complexes and finally results in the depletion of COL17 expression in basal keratinocytes.
• Mucosal hyperpigmentation from prophylactic minocycline for EGFR inhibitor

  K. Imafuku, K. Natsuga, S. Aoyagi, H. Shimizu

  JOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY AND VENEREOLOGY 30 (4) 690 - 692 0926-9959 2016/04 [Refereed][Not invited]
   

  Imafuku K, Natsuga K, Aoyagi S, Shimizu H, Journal of the European Academy of Dermatology and Venereology : JEADV, 2015
• Metal implant-induced skin ulcer mimicking scrofuloderma

  S. Kitamura, K. Natsuga, K. Imafuku, E. Homma, N. Yamane, S. Aoyagi, T. Matsumura, H. Shimizu

  JOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY AND VENEREOLOGY 30 (3) 449 - 450 0926-9959 2016/03 [Refereed][Not invited]
   

  Kitamura S, Natsuga K, Imafuku K, Homma E, Yamane N, Aoyagi S, Matsumura T, Shimizu H, Journal of the European Academy of Dermatology and Venereology : JEADV, 2014
• Concomitant neoplasms in the skin and stomach unveil the role of type IV collagen and E-cadherin in mucin core protein 5AC expression in vivo

  H. Hata, K. Natsuga, S. Kitamura, K. Imafuku, Y. Yamaguchi, Y. Ebihara, T. Shichinohe, S. Hirano, H. Shimizu

  BRITISH JOURNAL OF DERMATOLOGY 174 (2) 395 - 397 0007-0963 2016/02 [Refereed][Not invited]
   

  Mucin core protein (MUC) 5AC is a gel-forming glycoprotein that is expressed in different types of tumour cells. MUC5AC expression in cultured cells is regulated through the extracellular matrix and through remodelling by other membranous proteins such as type IV collagen (COL4) and E-cadherin. However, it has not been elucidated whether COL4 and E-cadherin affect MUC5AC expression in tumours in vivo. Here, by analysing a single individual with concomitant neoplasms in the skin [extramammary Paget disease (EMPD)] and the stomach (gastric cancer), we show that MUC5AC expression is reduced in COL4 and membranous E-cadherin-expressing EMPD specimens whereas MUC5AC is not abolished in gastric cancer with COL4 negativity and E-cadherin cytoplasmic localization. As the EMPD and gastric cancer specimens were derived from a single patient, each specimen had the same genetic background. These in vivo results support previous in vitro studies which showed that COL4 and E-cadherin down-regulated MUC5AC expression. Our study suggests that concomitant neoplasms in different organs of the same individual can serve as a strong tool for uncovering functional diversity in tumour markers in distinct cancer cells.
• Pyoderma Gangrenosum Associated with Acute Respiratory Distress Syndrome

  M. Watanabe, K. Natsuga, M. Ota, K. Ito

  American Journal of Medicine 129 (2) e17 - e18 1555-7162 2016/02/01 [Refereed][Not invited]
   

  Watanabe M, Natsuga K, Ota M, Ito K, The American journal of medicine, 2016, vol. 129, no. 2, pp. e17-8
• Epidermolysis Bullosa Acquisita Develops in Dominant Dystrophic Epidermolysis Bullosa

  Ryota Hayashi, Ken Natsuga, Mika Watanabe, Hiroaki Iwata, Satoru Shinkuma, Akiko Ito, Yukiko Masui, Masaaki Ito, Yutaka Shimomura

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 136 (1) 320 - 323 0022-202X 2016/01 [Refereed][Not invited]
   

  Hayashi R, Natsuga K, Watanabe M, Iwata H, Shinkuma S, Ito A, Masui Y, Ito M, Shimomura Y, The Journal of investigative dermatology, 2015
• Increased Bacterial Load and Expression of Antimicrobial Peptides in Skin of Barrier-Deficient Mice with Reduced Cancer Susceptibility

  Ken Natsuga, Sara Cipolat, Fiona M. Watt

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 136 (1) 99 - 106 0022-202X 2016/01 [Refereed][Not invited]
   

  Mice lacking three epidermal barrier proteins-envoplakin, periplakin, and involucrin (EPI-/- mice)-have a defective cornified layer, reduced epidermal gamma delta T cells, and increased dermal CD4(+) T cells. They are also resistant to developing skin tumors. The tumor-protective mechanism involves signaling between Rae-1 expressing keratinocytes and the natural killer group 2D receptor on immune cells, which also plays a role in host defenses against infection. Given the emerging link between bacteria and cancer, we investigated whether EPI-/- mice have an altered skin microbiota. The bacterial phyla were similar in wild-type and EPI-/- skin. However, bacteria were threefold more abundant in EPI-/- skin and penetrated deeper into the epidermis. The major epithelial defense mechanism against bacteria is production of antimicrobial proteins (AMPs). EPI-/- skin exhibited enhanced expression of antimicrobial peptides. However, reducing the bacterial load by antibiotic treatment or breeding mice under specific pathogen-free conditions did not reduce AMP expression or alleviate the abnormalities in T-cell populations. We conclude that the atopic characteristics of EPI-/- skin are a consequence of the defective barrier rather than a response to the increased bacterial load. It is therefore unlikely that the increase in skin microbiota contributes directly to the observed cancer resistance.
• Mucocutaneous pyoderma gangrenosum due to trisomy 8 neutrophilic infiltrates in a patient with myelodysplastic syndrome

  N. Haga, H. Iwata, Y. Yamaguchi, T. Shirato, K. Nishimura, N. Yamane, S. Shinkuma, K. Natsuga, T. Kondo, H. Shimizu

  BRITISH JOURNAL OF DERMATOLOGY 174 (1) 239 - 241 0007-0963 2016/01 [Refereed][Not invited]
   

  Haga N, Iwata H, Yamaguchi Y, Shirato T, Nishimura K, Yamane N, Shinkuma S, Natsuga K, Kondo T, Shimizu H, The British journal of dermatology, 2016, vol. 174, no. 1, pp. 239-241
• Anti-BP180-type mucous membrane pemphigoid: report of two cases

  Mayumi Wada, Jun Sato, Masanobu Shindoh, Hideyuki Ujiie, Ken Natsuga, Wataru Nishie, Hiroshi Shimizu, Yoshimasa Kitagawa

  ODONTOLOGY 104 (1) 114 - 118 1618-1247 2016/01 [Refereed][Not invited]
   

  We describe two patients with anti-BP180-type mucous membrane pemphigoid (MMP), who were correctly diagnosed and treated in early stages through the cooperation of dentists and dermatologists. Patient 1 was a 74-year-old woman who visited our dental department due to blisters over the oral mucosa and eruptions on the skin. She had also experienced bleeding of the gingiva and palate mucosa. Biopsy specimens from the oral mucosa revealed detachment of epithelial basement membrane and subepithelial lamina propria with slight chronic inflammation. Direct immunofluorescence (DIF) revealed linear IgG and IgA deposits along the basement membrane zone (BMZ). Indirect immunofluorescence (IIF) using 1 M-NaCl split normal human skin showed binding of IgG and IgA on the epidermal side. On immunoblot analysis, IgG and IgA autoantibodies reacted with the C-terminal protein of BP180. These findings indicated a diagnosis of anti-BP180-type MMP. Patient 2 was a 59-year-old woman who was referred to our dental department with a history of blisters and large erosions on the gingiva. Biopsy specimens from the oral mucosa revealed partial junctional separation at the level of the basement membrane. DIF showed linear depositions of IgG and C3 along the BMZ. IIF, using 1 M-NaCl split normal human skin, revealed circulating anti-BMZ-IgG antibodies bound to the epidermal side. These findings indicated a diagnosis of anti-BP180-type MMP. Both patients were treated successfully with systemic or topical steroids and oral health care. In conclusion, appropriate clinical examination and cooperation among medical specialists are important for the early diagnosis and treatment of patients with recurrent and chronic stomatitis and for their good prognosis.
• Anti-laminin-gamma 1 Pemphigoid with Generalized Pustular Psoriasis and Psoriasis Vulgaris

  Yu Fujimura, Ken Natsuga, Yohei Hamade, Yukiko Nomura, Yo Kaku, Ryuichi Muramatsu, Hiroshi Shimizu

  ACTA DERMATO-VENEREOLOGICA 96 (1) 120 - 121 0001-5555 2016 [Refereed][Not invited]
   

  Fujimura Y, Natsuga K, Hamade Y, Nomura Y, Kaku Y, Muramatsu R, Shimizu H, Acta dermato-venereologica, 2015
• Extracellular cleavage of collagen XVII is essential for correct cutaneous basement membrane formation

  Machiko Nishimura, Wataru Nishie, Yoshinori Shirafuji, Satoru Shinkuma, Ken Natsuga, Hideki Nakamura, Daisuke Sawamura, Keiji Iwatsuki, Hiroshi Shimizu

  HUMAN MOLECULAR GENETICS 25 (2) 328 - 339 0964-6906 2016/01 [Refereed][Not invited]
   

  In skin, basal keratinocytes in the epidermis are tightly attached to the underlying dermis by the basement membrane (BM). The correct expression of hemidesmosomal and extracellular matrix (ECM) proteins is essential for BM formation, and the null-expression of one molecule may induce blistering diseases associated with immature BM formation in humans. However, little is known about the significance of post-translational processing of hemidesmosomal or ECM proteins in BM formation. Here we show that the C-terminal cleavage of hemidesmosomal transmembrane collagen XVII (COL17) is essential for correct BM formation. The homozygous p.R1303Q mutation in COL17 induces BM duplication and blistering in humans. Although laminin 332, a major ECM protein, interacts with COL17 around p.R1303, the mutation leaves the binding of both molecules unchanged. Instead, the mutation hampers the physiological C-terminal cleavage of COL17 in the ECM. Consequently, non-cleaved COL17 ectodomain remnants induce the aberrant deposition of laminin 332 in the ECM, which is thought to be the major pathogenesis of the BM duplication that results from this mutation. As an example of impaired cleavage of COL17, this study shows that regulated processing of hemidesmosomal proteins is essential for correct BM organization in skin.
• Nodular morphoea: a first case associated with linear morphoea

  Shinichi Nakazato, Toshifumi Nomura, Naoko Yamane, Satoru Shinkuma, Ken Natsuga, Yasuyuki Fujita, Ken Arita, Hiroshi Shimizu

  EUROPEAN JOURNAL OF DERMATOLOGY 26 (1) 95 - 96 1167-1122 2016/01 [Refereed][Not invited]
   

  Nakazato S, Nomura T, Yamane N, Shinkuma S, Natsuga K, Fujita Y, Arita K, Shimizu H, European journal of dermatology : EJD, 2016
• Extensive Erythema and Hyperkeratosis on the Extremities and Lumbar Area as an Unusual Manifestation of Nagashima-type Palmoplantar Keratosis

  Toshinari Miyauchi, Toshifumi Nomura, Shotaro Suzuki, Yuka Ohg-Uchi, Yasuyuki Yamaguchi, Satoru Shinkuma, Ken Natsuga, Yasuyuki Fujita, Hiroshi Shimizu

  ACTA DERMATO-VENEREOLOGICA 96 (6) 856 - 858 0001-5555 2016 [Refereed][Not invited]
   

  Miyauchi T, Nomura T, Suzuki S, Ohguchi Y, Yamaguchi Y, Shinkuma S, Natsuga K, Fujita Y, Shimizu H, Acta dermato-venereologica, 2016
• Plasma cell cheilitis extending beyond vermillion border

  Yu Fujimura, Ken Natsuga, Riichiro Abe, Yusuke Morita, Toshifumi Nomura, Hiroshi Shimizu

  JOURNAL OF DERMATOLOGY 42 (9) 935 - 936 0385-2407 2015/09 [Refereed][Not invited]
   

  Fujimura Y, Natsuga K, Abe R, Morita Y, Nomura T, Shimizu H, The Journal of dermatology, 2015, vol. 42, no. 9, pp. 935-936
• Non-solar-induced elastotic bands on the forearm

  Naoya Haga, Riichiro Abe, Yusuke Morita, Yu Fujimura, Ken Natsuga, Toshifumi Nomura, Masanobu Kumakiri, Hiroshi Shimizu

  EUROPEAN JOURNAL OF DERMATOLOGY 25 (5) 508 - 509 1167-1122 2015/09 [Refereed][Not invited]
   

  Haga N, Abe R, Morita Y, Fujimura Y, Natsuga K, Nomura T, Kumakiri M, Shimizu H, European journal of dermatology : EJD, 2015, vol. 25, no. 5, pp. 508-509
• Context-Dependent Regulation of Collagen XVII Ectodomain Shedding in Skin

  Wataru Nishie, Ken Natsuga, Hiroaki Iwata, Kentaro Izumi, Hideyuki Ujiie, Ellen Toyonaga, Hiroo Hata, Hideki Nakamura, Hiroshi Shimizu

  AMERICAN JOURNAL OF PATHOLOGY 185 (5) 1361 - 1371 0002-9440 2015/05 [Refereed][Not invited]
   

  Pemphigoid is a common autoimmune blistering disorder in which autoantibodies target transmembrane collagen XVII (COL17), a component of hemidesmosomes in basal keratinocytes. The ectodomain of COL17 can be cleaved from the cell surface within the juxtamembranous extracellular NC16A domain, and, interestingly, certain autoantibodies of pemphigoid patients preferentially react with the shed ectodomain. These findings suggest that COL17 ectodomain shedding generates neoepitopes on the shed form; however, the regulatory mechanism of the shedding in in vivo skin and the pathogenicity of the neoepitope-targeting antibodies still are uncertain. To address these issues, we produced rabbit antibodies specifically reacting with N-terminal cleavage sites of the shed COL17 ectodomain. The antibodies showed that certain amounts of the human COL17 ectodomain are cleaved physiologically at Gln(525) in in vivo skin. In contrast, migrating human keratinocytes cleave COL17 at Leu(524) but not at Gln(525). The passive transfer of antibodies reacting with an N-terminal cleavage site of the mouse COL17 ectodomain into neonatal wild-type mice failed to induce blister formation, even though the antibodies bound to the dermal-epidermal junctions, indicating that cleavage site-specific antibodies have reduced or absent pathogenicity for blister formation. This study shows the ectodomain shedding of COL17 to be a physiological event in in vivo human skin that probably generates nonpathologic epitopes on the cleavage sites.
• Skipped exon in COL7A1 determines the clinical phenotypes of dystrophic epidermolysis bullosa

  E. Toyonaga, W. Nishie, M. Komine, S. Murata, S. Shinkuma, K. Natsuga, H. Nakamura, M. Ohtsuki, H. Shimizu

  BRITISH JOURNAL OF DERMATOLOGY 172 (4) 1141 - 1144 0007-0963 2015/04 [Refereed][Not invited]
   

  Toyonaga E, Nishie W, Komine M, Murata S, Shinkuma S, Natsuga K, Nakamura H, Ohtsuki M, Shimizu H, The British journal of dermatology, 2014
• In vivo analysis of IgE autoantibodies in bullous pemphigoid: A study of 100 cases

  Reine Moriuchi, Wataru Nishie, Hideyuki Ujiie, Ken Natsuga, Hiroshi Shimizu

  JOURNAL OF DERMATOLOGICAL SCIENCE 78 (1) 21 - 25 0923-1811 2015/04 [Refereed][Not invited]
   

  Background: Bullous pemphigoid (BP) is an acquired autoimmune blistering disease characterized by subepidermal blister formation, in vivo linear deposition of immunoglobulin G (IgG) and complements at the dermal-epidermal junction (DEJ). The circulating IgG autoantibodies are directed against two epidermal hemidesmosomal glycoproteins: BP180, also known as type XVII collagen (COL17), and BP230. In addition, recent studies have shown that IgE autoantibodies may be involved in the pathogenesis of BP, although in vivo IgE deposition in lesional skin has not been fully characterized in large numbers of BP patients. Objective: This study investigated the incidence of in vivo deposition of IgE autoantibodies at the DEJ in lesional skin from a large number of BP patients. Methods: Peri-lesional skin samples from 100 patients who met the clinical and histopathological criteria for BP were investigated by direct immunofluorescence for the deposition of autoantibodies and complement. Patients' sera were also investigated by enzyme-linked immunosorbent assay and indirect immunofluorescence. Results: 18% of BP patients were found to show IgE deposition at the DEJ. Disease severity, clinical course and outcome did not differ between IgE-positive and IgE-negative patients. In 3 IgE-positive cases, IgG was undetectable in vivo, and these cases showed atypical manifestations. Conclusion: The results of in vivo IgE deposition may not be useful in predicting the disease course of BP, although predominant IgE deposition could alter the pattern of clinical manifestations. (C) 2015 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.
• Plectin-related skin diseases

  Ken Natsuga

  JOURNAL OF DERMATOLOGICAL SCIENCE 77 (3) 139 - 145 0923-1811 2015/03 [Refereed][Invited]
   

  Plectin has been characterized as a linker protein that is expressed in many cell types and is distinctive in various isoforms in the N-terminus and around the rod domain due to complicated alternative splicing of PLEC, the gene encoding plectin. Plectin deficiency causes autosomal recessive epidermolysis bullosa simplex (EBS) with involvement of the skin and other organs, such as muscle and gastrointestinal tract, depending on the expression pattern of the defective protein. In addition, a point mutation in the rod domain of plectin leads to autosomal dominant EBS, called as EBS-Ogna. Plectin can be targeted by circulating autoantibodies in subepidermal autoimmune blistering diseases. This review summarizes plectin-related skin diseases, from congenital to autoimmune disorders. (C) 2014 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.
• Bowen's disease on the palm presents refractory skin erosion without erythematous plaque

  Shinya Kitamura, Hiroaki Iwata, Keisuke Imafuku, Hiroo Hata, Ken Natsuga, Satoru Aoyagi, Hiroshi Shimizu

  JOURNAL OF DERMATOLOGY 42 (2) 225 - 226 0385-2407 2015/02 [Refereed][Not invited]
   

  Kitamura S, Iwata H, Imafuku K, Hata H, Natsuga K, Aoyagi S, Shimizu H, The Journal of dermatology, 2015, vol. 42, no. 2, pp. 225-226
• Bullous Pemphigoid Autoantibodies Directly Induce Blister Formation without Complement Activation

  Hideyuki Ujiie, Tetsumasa Sasaoka, Kentaro Izumi, Wataru Nishie, Satoru Shinkuma, Ken Natsuga, Hideki Nakamura, Akihiko Shibaki, Hiroshi Shimizu

  JOURNAL OF IMMUNOLOGY 193 (9) 4415 - 4428 0022-1767 2014/11 [Refereed][Not invited]
   

  Complement activation and subsequent recruitment of inflammatory cells at the dermal/epidermal junction are thought to be essential for blister formation in bullous pemphigoid (BP), an autoimmune blistering disease induced by autoantibodies against type XVII collagen (COL17); however, this theory does not fully explain the pathological features of BP. Recently, the involvement of complement-independent pathways has been proposed. To directly address the question of the necessity of the complement activation in blister formation, we generated C3-deficient COL17-humanized mice. First, we show that passive transfer of autoantibodies from BP patients induced blister formation in neonatal C3-deficient COL17-humanized mice without complement activation. By using newly generated human and murine mAbs against the pathogenic noncollagenous 16A domain of COL17 with high (human IgG1, murine IgG2), low (murine IgG1), or no (human IgG4) complement activation abilities, we demonstrate that the deposition of Abs, and not complements, is relevant to the induction of blister formation in neonatal and adult mice. Notably, passive transfer of BP autoantibodies reduced the amount of COL17 in lesional mice skin, as observed in cultured normal human keratinocytes treated with the same Abs. Moreover, the COL17 depletion was associated with a ubiquitin/proteasome pathway. In conclusion, the COL17 depletion induced by BP autoantibodies, and not complement activation, is essential for the blister formation under our experimental system.
• BLIMP1 Is Required for Postnatal Epidermal Homeostasis but Does Not Define a Sebaceous Gland Progenitor under Steady-State Conditions

  Kai Kretzschmar, Denny L. Cottle, Giacomo Donati, Ming-Feng Chiang, Sven R. Quist, Harald P. Gollnick, Ken Natsuga, Kuo-I Lin, Fiona M. Watt

  STEM CELL REPORTS 3 (4) 620 - 633 2213-6711 2014/10 [Refereed][Not invited]
   

  B-lymphocyte-induced nuclear maturation protein 1 (BLIMP1) was previously reported to define a sebaceous gland (SG) progenitor population in the epidermis. However, the recent identification of multiple stem cell populations in the hair follicle junctional zone has led us to re-evaluate its function. We show, in agreement with previous studies, that BLIMP1 is expressed by postmitotic, terminally differentiated epidermal cells within the SG, interfollicular epidermis, and hair follicle. Epidermal overexpression of c-Myc results in loss of BLIMP1(+) cells, an effect modulated by androgen signaling. Epidermal-specific deletion of Blimp1 causes multiple differentiation defects in the epidermis in addition to SG enlargement. In culture, BLIMP1(+) sebocytes have no greater clonogenic potential than BLIMP1(+) sebocytes. Finally, lineage-tracing experiments reveal that, under steady-state conditions, BLIMP1-expressing cells do not divide. Thus, rather than defining a sebocyte progenitor population, BLIMP1 functions in terminally differentiated cells to maintain homeostasis in multiple epidermal compartments.
• Solitary fibrous tumour fluctuating in size with menstrual cycle

  H. Hata, K. Natsuga, S. Aoyagi, E. Homma, H. Shimizu

  CLINICAL AND EXPERIMENTAL DERMATOLOGY 39 (6) 753 - 755 0307-6938 2014/08 [Refereed][Not invited]
   

  Hata H, Natsuga K, Aoyagi S, Homma E, Shimizu H, Clinical and experimental dermatology, 2014, vol. 39, no. 6, pp. 753-755
• A recurrent 'hot spot' glycine substitution mutation, G2043R in COL7A1, induces dominant dystrophic epidermolysis bullosa associated with intracytoplasmic accumulation of pro-collagen

  Wataru Nishie, Ken Natsuga, Hideki Nakamura, Takamasa Ito, Ellen Toyonaga, Hidetsugu Sato, Hiroshi Shimizu

  JOURNAL OF DERMATOLOGICAL SCIENCE 1 75 (1) 69 - 71 0923-1811 2014/07 [Refereed][Not invited]
   

  Nishie W, Natsuga K, Nakamura H, Ito T, Toyonaga E, Sato H, Shimizu H, Journal of dermatological science, 2014, vol. 75, no. 1, pp. 69-71
• Ultrasound B-mode and elastographic findings of angiomatoid fibrous histiocytoma

  H. Hata, K. Natsuga, S. Aoyagi, E. Homma, H. Shimizu

  CLINICAL AND EXPERIMENTAL DERMATOLOGY 39 (4) 538 - 539 0307-6938 2014/06 [Refereed][Not invited]
   

  Hata H, Natsuga K, Aoyagi S, Homma E, Shimizu H, Clinical and experimental dermatology, 2014, vol. 39, no. 4, pp. 538-539
• Paper-Based ELISA for the Detection of Autoimmune Antibodies in Body Fluid-The Case of Bullous Pemphigoid

  Chao-Kai Hsu, Hsin-Yu Huang, Wan-Rung Chen, Wataru Nishie, Hideyuki Ujiie, Ken Natsuga, Shu-Ting Fan, Hsi-Kai Wang, Julia Yu-Yun Lee, Wei-Lun Tsai, Hiroshi Shimizu, Chao-Min Cheng

  ANALYTICAL CHEMISTRY 86 (9) 4605 - 4610 0003-2700 2014/05 [Refereed][Not invited]
   

  Bullous pemphigoid (BP), a common autoimmune blistering disease, is increasing in incidence and conveys a high mortality. Detection of autoantibodies targeting the noncollagenous 16A (NC16A) domain of type XVII collagen using enzyme-linked immunosorbent assay (ELISA) has demonstrated high sensitivity and specificity for diagnosing BP. We have developed a rapid, low-cost, and widely applicable ELISA-based system to detect the NC16A autoimmune antibody and then diagnose and monitor BP disease activity using a piece of filter paper, a wax-printer, and NC16A antigens. Both sera and/or blister fluids from 14 untreated BP patients were analyzed. The control group included healthy volunteers and patients with other blistering disorders such as pemphigus vulgaris. In our established paper-based ELISA (P-ELISA) system, only 2 mu L of serum or blister fluid and 70 min were required to detect anti-NC16A autoimmune antibodies. The relative color intensity was significantly higher in the BP group than in the control groups when using either serum (P < 0.05) or blister fluid (P < 0.001) specimens from BP patients. The results of P-ELISA were moderately correlated with the titer of the commercial ELISA kit (MBL, Japan) (rho = 0.5680, P = 0.0011). This newly developed system allows for rapid and convenient diagnosis and/or monitoring of BP disease activity.
• Epidermal barrier defects link atopic dermatitis with altered skin cancer susceptibility

  Sara Cipolat, Esther Hoste, Ken Natsuga, Sven R. Quist, Fiona M. Watt

  ELIFE 3 (3) e01888  2050-084X 2014/05 [Refereed][Not invited]
   

  Atopic dermatitis can result from loss of structural proteins in the outermost epidermal layers, leading to a defective epidermal barrier. To test whether this influences tumour formation, we chemically induced tumours in EPI-/- mice, which lack three barrier proteins-Envoplakin, Periplakin, and Involucrin. EPI-/- mice were highly resistant to developing benign tumours when treated with 7,12-dimethylbenz(a)anthracene (DMBA) and 12-O-tetradecanoylphorbol-13-acetate (TPA). The DMBA response was normal, but EPI-/- skin exhibited an exaggerated atopic response to TPA, characterised by abnormal epidermal differentiation, a complex immune infiltrate and elevated serum thymic stromal lymphopoietin (TSLP). The exacerbated TPA response could be normalised by blocking TSLP or the immunoreceptor NKG2D but not CD4+ T cells. We conclude that atopy is protective against skin cancer in our experimental model and that the mechanism involves keratinocytes communicating with cells of the immune system via signalling elements that normally protect against environmental assaults.
• Epidermal Barriers

  Ken Natsuga

  COLD SPRING HARBOR PERSPECTIVES IN MEDICINE 4 (4) a018218  2157-1422 2014/04 [Refereed][Invited]
   

  The epidermis functions as a physical barrier to the external environment and works to prevent loss of water from the skin. Numerous factors have been implicated in the formation of epidermal barriers, such as cornified envelopes, corneocytes, lipids, junctional proteins, proteases, protease inhibitors, antimicrobial peptides, and transcription factors. This review illustrates human diseases (ichthyoses) and animal models in which the epidermal barrier is disrupted or dysfunctional at steady state owing to ablation of one or more of the above factors. These diseases and animal models help us to understand the complicated mechanisms of epidermal barrier formation and give further insights on epidermal development.
• Epidermal Wnt/beta-catenin signaling regulates adipocyte differentiation via secretion of adipogenic factors

  Giacomo Donati, Valentina Proserpio, Beate Maria Lichtenberger, Ken Natsuga, Rodney Sinclair, Hironobu Fujiwara, Fiona M. Watt

  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 15 111 (15) E1501 - E1509 0027-8424 2014/04 [Refereed][Not invited]
   

  It has long been recognized that the hair follicle growth cycle and oscillation in the thickness of the underlying adipocyte layer are synchronized. Although factors secreted by adipocytes are known to regulate the hair growth cycle, it is unclear whether the epidermis can regulate adipogenesis. We show that inhibition of epidermal Wnt/beta-catenin signaling reduced adipocyte differentiation in developing and adult mouse dermis. Conversely, ectopic activation of epidermal Wnt signaling promoted adipocyte differentiation and hair growth. When the Wnt pathway was activated in the embryonic epidermis, there was a dramatic and premature increase in adipocytes in the absence of hair follicle formation, demonstrating that Wnt activation, rather than mature hair follicles, is required for adipocyte generation. Epidermal and dermal gene expression profiling identified keratinocyte-derived adipogenic factors that are induced by beta-catenin activation. Wnt/beta-catenin signaling-dependent secreted factors from keratinocytes promoted adipocyte differentiation in vitro, and we identified ligands for the bone morphogenetic protein and insulin pathways as proadipogenic factors. Our results indicate epidermal Wnt/beta-catenin as a critical initiator of a signaling cascade that induces adipogenesis and highlight the role of epidermal Wnt signaling in synchronizing adipocyte differentiation with the hair growth cycle.
• Dermoscopic Features of Plasma Cell Cheilitis and Actinic Cheilitis

  Takamasa Ito, Ken Natsuga, Shintaro Tanimura, Satoru Aoyagi, Hiroshi Shimizu

  ACTA DERMATO-VENEREOLOGICA 94 (5) 593 - 594 0001-5555 2014 [Refereed][Not invited]
   

  Ito T, Natsuga K, Tanimura S, Aoyagi S, Shimizu H, Acta dermato-venereologica, 2014, vol. 94, no. 5, pp. 593-594
• Coexistence case of bullous pemphigoid and pemphigus foliaceus

  Masumi Tsujiwaki, Riichiro Abe, Yukiko Nomura, Machiko Nishimura, Daichi Hoshina, Satoru Shinkuma, Ken Natsuga, Hideyuki Ujiie, Ken Arita, Hiroshi Shimizu

  EUROPEAN JOURNAL OF DERMATOLOGY 23 (4) 552 - 553 1167-1122 2013/07 [Refereed][Not invited]
   

  Tsujiwaki M, Abe R, Nomura Y, Nishimura M, Hoshina D, Shinkuma S, Natsuga K, Ujiie H, Arita K, Shimizu H, European journal of dermatology : EJD, 2013, vol. 23, no. 4, pp. 552-553
• C-MYC-Induced Sebaceous Gland Differentiation Is Controlled by an Androgen Receptor/p53 Axis

  Denny L. Cottle, Kai Kretzschmar, Pawel J. Schweiger, Sven R. Quist, Harald P. Gollnick, Ken Natsuga, Satoru Aoyagi, Fiona M. Watt

  Cell Reports 3 (2) 427 - 441 2211-1247 2013 [Refereed][Not invited]
   

  Although the sebaceous gland (SG) plays an important role in skin function, the mechanisms regulating SG differentiation and carcinoma formation are poorly understood. We previously reported that c-MYC overexpression stimulates SG differentiation. We now demonstrate roles for the androgen receptor (AR) and p53. MYC-induced SG differentiation was reduced in mice lacking a functional AR. High levels of MYC triggered a p53-dependent DNA damage response, leading to accumulation of proliferative SG progenitors and inhibition of AR signaling. Conversely, testosterone treatment or p53 deletion activated AR signaling and restored MYC-induced differentiation. Poorly differentiated human sebaceous carcinomas exhibited high p53 and low AR expression. Thus, the consequences of overactivating MYC in the SG depend on whether AR or p53 is activated, as they form a regulatory axis controlling proliferation and differentiation.
• A Novel Keratin 5 Mutation in an African Family with Epidermolysis Bullosa Simplex Indicates the Importance of the Amino Acid Located at the Boundary Site Between the H1 and Coil 1A Domains

  Satoru Shinkuma, Wataru Nishie, Witold K. Jacyk, Ken Natsuga, Hideyuki Ujiie, Hideki Nakamura, Masashi Akiyama, Hiroshi Shimizu

  ACTA DERMATO-VENEREOLOGICA 93 (5) 585 - 587 0001-5555 2013 [Refereed][Not invited]
   

  Shinkuma S, Nishie W, Jacyk WK, Natsuga K, Ujiie H, Nakamura H, Akiyama M, Shimizu H, Acta dermato-venereologica, 2013, vol. 93, no. 5, pp. 585-587
• The beta 9 Loop Domain of PA-PLA(1)alpha Has a Crucial Role in Autosomal Recessive Woolly Hair/Hypotrichosis

  Satoru Shinkuma, Asuka Inoue, Junken Aoki, Wataru Nishie, Ken Natsuga, Hideyuki Ujiie, Toshifumi Nomura, Riichiro Abe, Masashi Akiyama, Hiroshi Shimizu

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 8 132 (8) 2093 - 2095 0022-202X 2012/08 [Refereed][Not invited]
   

  Shinkuma S, Inoue A, Aoki J, Nishie W, Natsuga K, Ujiie H, Nomura T, Abe R, Akiyama M, Shimizu H, The Journal of investigative dermatology, 2012, vol. 132, no. 8, pp. 2093-2095
• Antibodies to Pathogenic Epitopes or Type XVII Collagen Cause Skin Fragility in a Complement-Dependent and -Independent Manner

  Ken Natsuga, Wataru Nishie, Satoru Shinkuma, Hideyuki Ujiie, Machiko Nishimura, Daisuke Sawamura, Hiroshi Shimizu

  JOURNAL OF IMMUNOLOGY 188 (11) 5792 - 5799 0022-1767 2012/06 [Refereed][Not invited]
   

  In bullous pemphigoid (BP), the most prevalent autoimmune blistering disease, type XVII collagen (COL17) is targeted by circulating autoantibodies. BP is thought to be an autoantibody-mediated complement-fixing blistering disease, and a juxtamembranous noncollagenous 16A (NC16A) domain spanning Glu(490) to Arg(566) was proved to be the main pathogenic region on COL17, although precise pathogenic epitopes within NC16A have not been elucidated. In this study, we showed that injection of rabbit IgG Abs targeting Asp(522) to Gln(545) induced skin fragility associated with in vivo deposition of IgG and complement in neonatal COL17-humanized mice. Notably, immunoadsorption of rabbit anti-NC16A IgG Ab with this epitope (Asp(522) to Gln(545)) or the anti-NC16A IgG administered together with the peptides of this epitope as a decoy ameliorated skin fragility in the injected neonatal COL17-humanized mice compared with the anti-NC16A IgG alone even though all of the mice showed both IgG and complement deposition. These results led us to investigate an additional, complement-independent mechanism of skin fragility in the mice injected with anti-COL17 Abs. The rabbit anti-NC16A IgG depleted the expression of COL17 in cultured normal human keratinocytes, whereas immunoadsorption of the same IgG with this epitope significantly suppressed the depletion effect. Moreover, passive transfer of F(ab')(2) fragments of the human BP or rabbit IgG Abs against COL17 demonstrated skin fragility in neonatal COL17-humanized mice. In summary, this study reveals the importance of Abs directed against distinct epitopes on COL17, which induce skin fragility in complement-dependent as well as complement-independent ways. The Journal of Immunology, 2012, 188: 5792-5799.
• Mucous membrane pemphigoid with generalized blisters: IgA and IgG autoantibodies target both laminin-332 and type XVII collagen

  I. Hayashi, S. Shinkuma, S. Shimizu, K. Natsuga, H. Ujiie, C. Yasui, K. Tsuchiya, W. Nishie, H. Shimizu

  BRITISH JOURNAL OF DERMATOLOGY 166 (5) 1116 - 1120 0007-0963 2012/05 [Refereed][Not invited]
   

  Mucous membrane pemphigoid (MMP) is a mucous membrane-dominated, subepidermal autoimmune blistering disease in which autoantibodies usually react with the C-terminal domain of type XVII collagen (COL17) or with laminin-332. Only a few cases of MMP with widespread blisters have been reported. Serologically, IgA and IgG class autoantibodies directed against COL17 or IgG autoantibodies directed against laminin-332 in patients with MMP have been well documented. MMP cases in which IgA reacts with laminin-332, however, are extremely rare. We report a case of MMP in a 67-year-old man. Clinical examination revealed extensive mucosal lesions as well as generalized blisters and erosions that healed with scar formation. The disease was intractable to treatment with systemic steroids. Interestingly, in addition to IgG directed against laminin-332 and the noncollagenous 16A (NC16A) and C-terminal domains of COL17, circulating IgA reacting with laminin-332 and with the NC16A domain of COL17 was also detected. This is the first MMP case with circulating IgA and IgG autoantibodies against both laminin-332 and COL17.
• Possible modifier effects of keratin 17 gene mutation on keratitis-ichthyosis-deafness syndrome

  K. Natsuga, S. Shinkuma, M. Kanda, Y. Suzuki, N. Chosa, Y. Narita, M. Setoyama, W. Nishie, M. Akiyama, H. Shimizu

  BRITISH JOURNAL OF DERMATOLOGY 166 (4) 903 - 905 0007-0963 2012/04 [Refereed][Not invited]
   

  Natsuga K, Shinkuma S, Kanda M, Suzuki Y, Chosa N, Narita Y, Setoyama M, Nishie W, Akiyama M, Shimizu H, The British journal of dermatology, 2012, vol. 166, no. 4, pp. 903-905
• Subepidermal blistering disease with 3 distinct autoantibodies: Anti-BP230, anti-laminin gamma-1, and anti-laminin-332

  Kazuhiro Kikuchi, Ken Natsuga, Satoru Shinkuma, Wataru Nishie, Satoshi Kajita, Hidetsugu Sato, Hiroshi Shimizu

  JOURNAL OF THE AMERICAN ACADEMY OF DERMATOLOGY 65 (4) 878 - 880 0190-9622 2011/10 [Refereed][Not invited]
   

  Kikuchi K, Natsuga K, Shinkuma S, Nishie W, Kajita S, Sato H, Shimizu H, Journal of the American Academy of Dermatology, 2011, vol. 65, no. 4, pp. 878-880
• Consequences of Two Different Amino-Acid Substitutions at the Same Codon in KRT14 Indicate Definitive Roles of Structural Distortion in Epidermolysis Bullosa Simplex Pathogenesis

  Ken Natsuga, Wataru Nishie, Brian J. Smith, Satoru Shinkuma, Thomasin A. Smith, David A. D. Parry, Naoki Oiso, Akira Kawada, Kozo Yoneda, Masashi Akiyama, Hiroshi Shimizu

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 131 (9) 1869 - 1876 0022-202X 2011/09 [Refereed][Not invited]
   

  Numerous inherited diseases develop due to missense mutations, leading to an amino-acid substitution. Whether an amino-acid change is pathogenic depends on the level of deleterious effects caused by the amino-acid alteration. We show an example of different structural and phenotypic consequences caused by two individual amino-acid changes at the same position. Epidermolysis bullosa simplex (EBS) is a genodermatosis resulting from KRT5 or KRT14 mutations. Mutation analysis of an EBS family revealed that affected individuals were heterozygous for a, to our knowledge, previously unreported mutation of c.1237G4C (p.Ala413Pro) in KRT14. Interestingly, 2 of 100 unrelated normal controls were heterozygous, and 1 of the 100 was homozygous for a different mutation in this position, c.1237G4A (p.Ala413Thr). In silico modeling of the protein demonstrated deleterious structural effects from proline substitution but not from threonine substitution. In vitro transfection studies revealed a significantly larger number of keratin-clumped cells in HaCaT cells transfected with mutant KRT14 complementary DNA (cDNA) harboring p.Ala413Pro than those transfected with wild-type KRT14 cDNA or mutant KRT14 cDNA harboring p. Ala413Thr. These results show that changes in two distinct amino acids at a locus are destined to elicit different phenotypes due to the degree of structural distortion resulting from the amino-acid alterations.
• Malignant skin tumours in patients with inherited ichthyosis

  K. Natsuga, M. Akiyama, H. Shimizu

  BRITISH JOURNAL OF DERMATOLOGY 165 (2) 263 - 268 0007-0963 2011/08 [Refereed][Not invited]
   

  Inherited ichthyoses are rare genodermatoses caused by mutations in the genes involved in epidermal development. Although there have been case reports on patients with ichthyosis who developed skin malignancies, it is still unknown whether or not patients with ichthyosis have an increased risk of skin malignancies. Here, we review case series of skin malignancies in patients with ichthyosis and show biological findings which might lead to cancer susceptibility. A survey of the literature revealed 28 cases of inherited ichthyoses with skin malignancy, including 12 cases of keratitis-ichthyosis-deafness (KID) syndrome, seven of autosomal recessive congenital ichthyosis, three of Netherton syndrome and six of miscellaneous ichthyosis. Twenty-four of the 28 cases developed single or multiple squamous cell carcinomas (SCCs). The age at diagnosis of the first skin malignancy ranged from 15 to 54 years. As patients with these particular sub-types of ichthyosis seem to be prone to skin malignancies, including SCC, at an unusually young age, routine cancer surveillance of these patients is strongly recommended.
• Spontaneous remission of solitary-type infantile myofibromatosis.

  Kazuhiro Kikuchi, Riichiro Abe, Satoru Shinkuma, Erika Hamasaka, Ken Natsuga, Hiroo Hata, Yasuki Tateishi, Masahiko Shibata, Yuki Tomita, Yukiko Abe, Satoru Aoyagi, Makio Mukai, Hiroshi Shimizu

  Case reports in dermatology 3 (2) 181 - 5 1662-6567 2011/05 [Refereed][Not invited]
   

  Infantile myofibromatosis is a rare fibrous tumor of infancy. The cutaneous solitary type has typically an excellent prognosis. However, histologically, it is important to rule out leiomyosarcoma, which has a poor prognosis. The low frequency of mitosis was definitive for a diagnosis of infantile myofibromatosis. We present a cutaneous solitary-type case of infantile myofibromatosis. Following incisional biopsy, the tumor remitted spontaneously.
• DNA-based prenatal diagnosis of plectin-deficient epidermolysis bullosa simplex associated with pyloric atresia

  Hideki Nakamura, Ken Natsuga, Wataru Nishie, James R. McMillan, Hiroyuki Nakamura, Daisuke Sawamura, Masashi Akiyama, Hiroshi Shimizu

  INTERNATIONAL JOURNAL OF DERMATOLOGY 50 (4) 439 - 442 0011-9059 2011/04 [Refereed][Not invited]
   

  Background Mutations in the plectin gene (PLEC) generally lead to epidermolysis bullosa simplex (EBS) associated with muscular dystrophy. It has been recently demonstrated that PLEC mutations can also cause a different clinical subtype, EBS associated with pyloric atresia (EBS-PA), which shows early lethality. Prenatal diagnosis (PND) of EBS-PA using mutation screening of PLEC has not been described. Objective This study aimed to perform DNA-based PND for an EBS-PA family. Materials and methods The EBS-PA proband was compound-heterozygous for a paternal c.1350G > A splice-site mutation and a maternal p.Q305X nonsense mutation. Genomic DNA was obtained from amniocytes taken from an at-risk fetus of the proband's family. Direct sequencing and restriction enzyme digestion of polymerase chain reaction products from the genomic DNA were performed. Results Mutational analysis showed that the fetus harbored both pathogenic mutations, suggesting that the fetus was a compound-heterozygote and therefore affected with EBS-PA. The skin sample obtained by autopsy from the abortus confirmed the absence of plectin expression at the dermal-epidermal junction. Conclusions This is the first successful DNA-based PND for an EBA-PA family.
• Childhood subepidermal blistering disease with autoantibodies to type VII collagen and laminin-332

  H. -Y. Lin, T. Yanagi, M. Akiyama, M. M. Iitani, R. Moriuchi, K. Natsuga, S. Shinkuma, N. Yamane, D. Inokuma, K. Arita, H. Shimizu

  BRITISH JOURNAL OF DERMATOLOGY 164 (2) 452 - 454 0007-0963 2011/02 [Refereed][Not invited]
   

  Lin HY, Yanagi T, Akiyama M, Iitani MM, Moriuchi R, Natsuga K, Shinkuma S, Yamane N, Inokuma D, Arita K, Shimizu H, The British journal of dermatology, 2011, vol. 164, no. 2, pp. 452-454
• Type XVII collagen ELISA indices significantly decreased after bullous pemphigoid remission

  Erika Kusajima, Masashi Akiyama, Megumi Sato, Ken Natsuga, Hiroshi Shimizu

  INTERNATIONAL JOURNAL OF DERMATOLOGY 50 (2) 238 - 240 0011-9059 2011/02 [Refereed][Not invited]
   

  Kusajima E, Akiyama M, Sato M, Natsuga K, Shimizu H, International journal of dermatology, 2011, vol. 50, no. 2, pp. 238-240
• A founder effect of c.1938delC in ITGB4 underlies junctional epidermolysis bullosa and its application for prenatal testing

  Ken Natsuga, Wataru Nishie, Satoru Shinkuma, Hideki Nakamura, Ken Arita, Kozo Yoneda, Takashi Kusaka, Toshihiro Yanagihara, Rika Kosaki, Haruhiko Sago, Masashi Akiyama, Hiroshi Shimizu

  EXPERIMENTAL DERMATOLOGY 20 (1) 74 - 76 0906-6705 2011/01 [Refereed][Not invited]
   

  Junctional epidermolysis bullosa associated with pyloric atresia (JEB-PA) is one of the most severe inherited skin diseases, characterized by generalized blister formation and occlusion of the pylorus at birth. Most JEB-PA patients have mutations in the gene encoding beta 4 integrin (ITGB4). No recurrent mutations in ITGB4 have been described as having founder effects. We collected three JEB-PA families with c.1938delC in ITGB4. Haplotype analysis using single nucleotide polymorphism markers throughout ITGB4 suggested one rare haplotype (2.8% of the Han Chinese and ethnic Japanese populations) in all alleles with c.1938delC. The parents of one of the three families sought prenatal diagnosis for a subsequent pregnancy. We succeeded in performing prenatal exclusion of JEB-PA using the foetal genomic DNA. Our study clearly demonstrated that recurrent c.1938delC in ITGB4 is a founder mutation in JEB-PA patients, and that genotyping of the mutation can be utilized for prenatal diagnosis of JEB-PA.
• Expression of exon-8-skipped kindlin-1 does not compensate for defects of Kindler syndrome

  Ken Natsuga, Wataru Nishie, Satoru Shinkuma, Hideki Nakamura, Yoichiro Matsushima, Aya Tatsuta, Mayumi Kornine, Hiroshi Shimizu

  JOURNAL OF DERMATOLOGICAL SCIENCE 61 (1) 38 - 44 0923-1811 2011/01 [Refereed][Not invited]
   

  Background: Kindler syndrome (KS) is a rare, inherited skin disease characterized by blister formation and generalized poikiloderma. Mutations in KIND1, which encodes kindlin-1, are responsible for KS. c.1089del/1089+1del is a recurrent splice-site deletion mutation in KS patients. Objective: To elucidate the effects of c.1089del/1089+1del at the mRNA and protein level. Methods: Two KS patients with c.1089del/1089+1del were included in this study. Immunofluorescence analysis of KS skin samples using antibodies against the dermo-epidermal junction proteins was performed. Exon-trapping experiments were performed to isolate the mRNA sequences transcribed from genomic DNA harbouring c.1089del/1089+1del. beta 1 integrin activation in HeLa cells transfected with truncated KIND1 cDNA was analyzed. Results: Immunofluorescence study showed positive expression of kindlin-1 in KS skin with c.1089del/1089+1del mutation. We identified the exon-8-skipped in-frame transcript as the main product among multiple splicing variants derived from that mutation. HeLa cells transfected with KIND1 cDNA without exon 8 showed impaired beta 1 integrin activation. Exon-8-coding amino acids are located in the FERM F2 domain, which is conserved among species, and the unstructured region between F2 and the pleckstrin homology domain. Conclusion: This study suggests that exon-8-skipped truncated kindlin-1 is functionally defective and does not compensate for the defects of KS, even though kindlin-1 expression in skin is positive. (c) 2010 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.
• Human IgG1 Monoclonal Antibody against Human Collagen 17 Noncollagenous 16A Domain Induces Blisters via Complement Activation in Experimental Bullous Pemphigoid Model

  Qiang Li, Hideyuki Ujiie, Akihiko Shibaki, Gang Wang, Reine Moriuchi, Hong-jiang Qiao, Hiroshi Morioka, Satoru Shinkuma, Ken Natsuga, Heather A. Long, Wataru Nishie, Hiroshi Shimizu

  JOURNAL OF IMMUNOLOGY 185 (12) 7746 - 7755 0022-1767 2010/12 [Refereed][Not invited]
   

  Bullous pemphigoid (BP) is an autoimmune blistering disease caused by IgG autoantibodies targeting the noncollagenous 16A (NC16A) domain of human collagen 17 (hCOL17), which triggers blister formation via complement activation. Previous in vitro analysis demonstrated that IgG1 autoantibodies showed much stronger pathogenic activity than IgG4 autoantibodies; however, the exact pathogenic role of IgG1 autoantibodies has not been fully demonstrated in vivo. We constructed a recombinant IgG1 mAb against hCOL17 NC16A from BP patients. In COL17-humanized mice, this mAb effectively reproduced a BP phenotype that included subepidermal blisters, deposition of IgG1, C1q and C3, neutrophil infiltration, and mast cell degranulation. Subsequently, alanine substitutions at various C1q binding sites were separately introduced to the Fc region of the IgG1 mAb. Among these mutated mAbs, the one that was mutated at the P331 residue completely failed to activate the complement in vitro and drastically lost pathogenic activity in COL17-humanized mice. These findings indicate that P331 is a key residue required for complement activation and that IgG1-dependent complement activation is essential for blister formation in BP. This study is, to our knowledge, the first direct evidence that IgG1 Abs to hCOL17 NC16A can induce blister formation in vivo, and it raises the possibility that IgG1 mAbs with Fc modification may be used to block pathogenic epitopes in autoimmune diseases. The Journal of Immunology, 2010, 185: 7746-7755.
• Complete Paternal Isodisomy of Chromosome 17 in Junctional Epidermolysis Bullosa with Pyloric Atresia

  Ken Natsuga, Wataru Nishie, Ken Arita, Satoru Shinkuma, Hideki Nakamura, Shogo Kubota, Sumihisa Imakado, Masashi Akiyama, Hiroshi Shimizu

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 130 (11) 2671 - 2674 0022-202X 2010/11 [Refereed][Not invited]
   

  Natsuga K, Nishie W, Arita K, Shinkuma S, Nakamura H, Kubota S, Imakado S, Akiyama M, Shimizu H, The Journal of investigative dermatology, 2010, vol. 130, no. 11, pp. 2671-2674
• Plectin Deficiency Leads to Both Muscular Dystrophy and Pyloric Atresia in Epidermolysis Bullosa Simplex

  Ken Natsuga, Wataru Nishie, Satoru Shinkuma, Ken Arita, Hideki Nakamura, Makiko Ohyama, Hitoshi Osaka, Takeshi Kambara, Yoshiaki Hirako, Hiroshi Shimizu

  HUMAN MUTATION 31 (10) E1687 - E1698 1059-7794 2010/10 [Refereed][Not invited]
   

  Plectin is a cytoskeletal linker protein which has a long central rod and N- and C-terminal globular domains. Mutations in the gene encoding plectin (PLEC) cause two distinct autosomal recessive subtypes of epidermolysis bullosa: EB simplex (EBS) with muscular dystrophy (EBS-MD), and EBS with pyloric atresia (EBS-PA). Previous studies have demonstrated that loss of full-length plectin with residual expression of the rodless isoform leads to EBS-MD, whereas complete loss or marked attenuation of expression of full-length and rodless plectin underlies the more severe EBS-PA phenotype. However, muscular dystrophy has never been identified in EBS-PA, not even in the severe form of the disease. Here, we report the first case of EBS associated with both pyloric atresia and muscular dystrophy. Both of the premature termination codon-causing mutations of the proband are located within exon 32, the last exon of PLEC. Immunofluorescence and immunoblot analysis of skin samples and cultured fibroblasts from the proband revealed truncated plectin protein expression in low amounts. This study demonstrates that plectin deficiency can indeed lead to both muscular dystrophy and pyloric atresia in an individual EBS patient. (C) 2010 Wiley-Liss, Inc.
• Bone marrow transplantation restores epidermal basement membrane protein expression and rescues epidermolysis bullosa model mice

  Yasuyuki Fujita, Riichiro Abe, Daisuke Inokuma, Mikako Sasaki, Daichi Hoshina, Ken Natsuga, Wataru Nishie, James R. McMillan, Hideki Nakamura, Tadamichi Shimizu, Masashi Akiyama, Daisuke Sawamura, Hiroshi Shimizu

  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 107 (32) 14345 - 14350 0027-8424 2010/08 [Refereed][Not invited]
   

  Attempts to treat congenital protein deficiencies using bone marrow-derived cells have been reported. These efforts have been based on the concepts of stem cell plasticity. However, it is considered more difficult to restore structural proteins than to restore secretory enzymes. This study aims to clarify whether bone marrow transplantation (BMT) treatment can rescue epidermolysis bullosa (EB) caused by defects in keratinocyte structural proteins. BMT treatment of adult collagen XVII (Col17) knockout mice induced donor-derived keratinocytes and Col17 expression associated with the recovery of hemidesmosomal structure and better skin manifestations, as well improving the survival rate. Both hematopoietic and mesenchymal stem cells have the potential to produce Col17 in the BMT treatment model. Furthermore, human cord blood CD34(+) cells also differentiated into keratinocytes and expressed human skin component proteins in transplanted immunocompromised (NOD/SCID/gamma(null)(c)) mice. The current conventional BMT techniques have significant potential as a systemic therapeutic approach for the treatment of human EB.
• Prevalent LIPH Founder Mutations Lead to Loss of P2Y5 Activation Ability of PA-PLA(1)alpha in Autosomal Recessive Hypotrichosis

  Satoru Shinkuma, Masashi Akiyama, Asuka Inoue, Junken Aoki, Ken Natsuga, Toshifumi Nomura, Ken Arita, Riichiro Abe, Kei Ito, Hideki Nakamura, Hideyuki Ujiie, Akihiko Shibaki, Hiraku Suga, Yuichiro Tsunemi, Wataru Nishie, Hiroshi Shimizu

  HUMAN MUTATION 31 (5) 602 - 610 1059-7794 2010/05 [Refereed][Not invited]
   

  Autosomal recessive hypotrichosis (ARH) is characterized by sparse hair on the scalp without other abnormalities. Three genes, DSG4, LIPH, and LPAR6 (P2RY5), have been reported to underlie ARH. We performed a mutation search for the three candidate genes in five independent Japanese ARH families and identified two LIPH mutations: c.736T > A (p.Cys246Ser) in all five families, and c.742C > A (p.His248Asn) in four of the five families. Out of 200 unrelated control alleles, we detected c.736T > A in three alleles and c.742C > A in one allele. Haplotype analysis revealed each of the two mutant alleles is derived from a respective founder. These results suggest the LIPH mutations are prevalent founder mutations for ARH in the Japanese population. LIPH encodes PA-PLA(1)alpha (LIPH), a membrane-associated phosphatidic acid-preferring phospholipase A(1)alpha. Two residues, altered by these mutations, are conserved among PA-PLA(1)alpha of diverse species. Cys(246) forms intramolecular disulfide bonds on the lid domain, a crucial structure for substrate recognition, and His(248) is one amino acid of the catalytic triad. Both p.Cys246Ser- and p.His248Asn-PA-PLA(1)alpha mutants showed complete abolition of hydrolytic activity and had no P2Y5 activation ability. These results suggest defective activation of P2Y5 due to reduced 2-acyl lysophosphatidic acid production by the mutant PA-PLA(1)alpha is involved in the pathogenesis of ARH. Hum Mutat 31:602-610, 2010. (C) 2010 Wiley-Liss, Inc.
• Epidermolysis Bullosa in Japan

  Satoru Shinkuma, Ken Natsuga, Wataru Nishie, Hiroshi Shimizu

  DERMATOLOGIC CLINICS 28 (2) 431 - + 0733-8635 2010/04 [Refereed][Not invited]
   

  Epidermolysis bullosa (EB) is a group of hereditary disorders characterized by mechanical stress-induced blistering of the skin and mucous membranes. This article discusses the prevalence among and genetic studies of Japanese patients with EB.
• Circulating IgA and IgE autoantibodies in antilaminin-332 mucous membrane pemphigoid

  K. Natsuga, W. Nishie, S. Shinkuma, R. Moriuchi, M. Shibata, M. Nishimura, T. Hashimoto, H. Shimizu

  BRITISH JOURNAL OF DERMATOLOGY 162 (3) 513 - 517 0007-0963 2010/03 [Refereed][Not invited]
   

  Background Antilaminin-332 mucous membrane pemphigoid (MMP) is a chronic autoimmune bullous disease that is often associated with internal malignancy. IgG autoantibodies against laminin-332 in patients with MMP are well documented; however, IgA and IgE autoantibodies against laminin-332 have not yet been described. Objectives To characterize IgA and IgE autoantibodies binding to laminin-332 in sera from patients with antilaminin-332 MMP. Methods Sera and skin samples from four patients who met the following criteria were used: (i) subepidermal blistering lesions present on the mucous membranes; (ii) in vivo deposition of IgG along the epidermal basement membrane zone of sampled skin; (iii) circulating IgG antibasement membrane zone antibodies that react with the dermal side of salt-split normal human skin; and (iv) circulating IgG autoantibodies that do not show positivity against type VII collagen or 200-kDa protein (p200 antigen) in immunoblot analysis using dermal extracts. Circulating IgG/IgA/IgE class autoantibodies against laminin-332 were determined by immunoblotting. Results Circulating IgG autoantibodies against the gamma 2, alpha 3/gamma 2, alpha 3 and alpha 3/beta 3/gamma 2 subunits of laminin-332 were demonstrated in sera from four patients, respectively. Serum from one of the four patients showed IgA reactivity with the alpha 3/beta 3/gamma 2 subunits of laminin-332. Serum from one of the four patients showed IgE reactivity with the gamma 2 subunit of laminin-332. The control sera failed to display IgG/IgA/IgE reactivity to laminin-332. Conclusions In addition to IgG autoantibodies, circulating IgA and IgE autoantibodies against laminin-332 are detectable in a subset of patients with antilaminin-332 MMP.
• Plectin Expression Patterns Determine Two Distinct Subtypes of Epidermolysis Bullosa Simplex

  Ken Natsuga, Wataru Nishie, Masashi Akiyama, Hideki Nakamura, Satoru Shinkuma, James R. McMillan, Akari Nagasaki, Cristina Has, Takeshi Ouchi, Akira Ishiko, Yoshiaki Hirako, Katsushi Owaribe, Daisuke Sawamura, Leena Bruckner-Tuderman, Hiroshi Shimizu

  HUMAN MUTATION 31 (3) 308 - 316 1059-7794 2010/03 [Refereed][Not invited]
   

  Plectin is a cytoskeletal linker protein that has a dumbbell-like structure with a long central rod and N- and C-terminal globular domains. Mutations in the gene encoding plectin (PLEC1) cause two distinct autosomal recessive subtypes of epidermolysis bullosa (EB): EB simplex with muscular dystrophy (EBS-MD), and EB simplex with pyloric atresia (EBS-PA). Here, we demonstrate that normal human fibroblasts express two different plectin isoforms including full-length and rodless forms of plectin. We performed detailed analysis of plectin expression patterns in six EBS-MD and three EBS-PA patients. In EBS-PA, expression of all plectin domains was found to be markedly attenuated or completely lost; in EBS-MD, the expression of the N- and C-terminal domains of plectin remained detectable, although the expression of rod domains was absent or markedly reduced. Our data suggest that loss of the full-length plectin isoform with residual expression of the rodless plectin isoform leads to EBS-MD, and that complete loss or marked attenuation of full-length and rodless plectin expression underlies the more severe EBS-PA phenotype. These results also clearly account for the majority of EBS-MD PLEC1 mutation restriction within the large exon 31 that encodes the plectin rod domain, whereas EBS-PA PLEC1 mutations are generally outside exon 31. Hum Mutat 31:308-316, 2010. (C) 2010 Wiley-Liss, Inc.
• Animal Models of Epidermolysis Bullosa

  Ken Natsuga, Satoru Shinkuma, Wataru Nishie, Hiroshi Shimizu

  DERMATOLOGIC CLINICS 28 (1) 137 - + 0733-8635 2010/01 [Refereed][Invited]
   

  For more than 2 decades, animal models have been used to clarify the pathogenic mechanisms of human diseases and develop new therapeutics for these diseases. Several therapies for human diseases have become available through trials using animal models. Epidermolysis bullosa (EB) is one of the most severe inherited skin disorders, whose effective treatments have not been fully available. EB is characterized by abnormalities of the proteins that consist of the dermoepidermal junction. EB has been classified into three major subtypes according to the level of skin cleavage: EB simplex, junctional EB, and dystrophic EB. To date, 13 genes have been shown to cause EB phenotype. After the discovery of the causative genes responsible for each EB subtype, many researchers have tried to develop EB animal models by genetically manipulating the corresponding genes.
• Response of Intractable Skin Ulcers in Recessive Dystrophic Epidermolysis Bullosa Patients to an Allogeneic Cultured Dermal Substitute

  Ken Natsuga, Daisuke Sawamura, Maki Goto, Erina Homma, Yuka Goto-Ohguchi, Satoru Aoyagi, Masashi Akiyama, Yoshimitsu Kuroyanagi, Hiroshi Shimizu

  ACTA DERMATO-VENEREOLOGICA 90 (2) 165 - 169 0001-5555 2010 [Refereed][Not invited]
   

  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.
• Localized Linear IgA/IgG Bullous Dermatosis

  Satoko Shimizu, Ken Natsuga, Satoru Shinkuma, Chikako Yasui, Kikuo Tsuchiya, Hiroshi Shimizu

  ACTA DERMATO-VENEREOLOGICA 90 (6) 621 - 624 0001-5555 2010 [Refereed][Not invited]
   

  Linear IgA/IgG bullous dermatosis (LAGBD) is an autoimmune blistering disease characterized by the local accumulation of IgA- and IgG-class anti-basement membrane autoantibodies It typically presents as a generalized pruritic vesiculobullous eruption No cases of localized LAGBD have yet been reported We report a case of a 78-year-old man with LAGBD localized to the perianal area The patient complained of suffering from persistent ulcers around the anus for more than 3 years Physical examination revealed several blisters and ulcers up to 2-cm in diameter around the anus No lesions were found elsewhere on the body Histological analysis of a skin biopsy revealed subepidermal blistering, while direct immunofluorescence showed the linear deposition of IgA and IgG antibodies at the dermoepidermal Junction Indirect immunofluorescence of normal human skin whose layers had been separated using 1M NaCl showed the binding of both IgA and IgG to the epidermal side Immunoblotting demonstrated the presence of circulating IgA and IgG autoantibodies that bound to a 120-kDa protein This is the first case of localized LAGBD whose skin lesions were restricted to the perianal region
• Keratinocyte-/Fibroblast-Targeted Rescue of Col7a1-Disrupted Mice and Generation of an Exact Dystrophic Epidermolysis Bullosa Model Using a Human COL7A1 Mutation

  Kei Ito, Daisuke Sawamura, Maki Goto, Hideki Nakamura, Wataru Nishie, Kaori Sakai, Ken Natsuga, Satoru Shinkuma, Akihiko Shibaki, Jouni Uitto, Christopher P. Denton, Osamu Nakajima, Masashi Akiyama, Hiroshi Shimizu

  AMERICAN JOURNAL OF PATHOLOGY 175 (6) 2508 - 2517 0002-9440 2009/12 [Refereed][Not invited]
   

  Recessive dystrophic epidermolysis bullosa (RDEB) is a severe hereditary bullous disease caused by mutations in COL7A1, which encodes type VU collagen (COL7). coral knockout mice (COL7(m-/-)) exhibit a severe RDEB phenotype and die within a few days after birth. Toward developing novel approaches for treating patients with RDEB, we attempted to rescue COL7(m-/-) mice by introducing human COL7A1 cDNA. we first generated transgenic mice that express human COL7A1 cDNA specifically in either epidermal keratinocytes or dermal fibroblasts. We then performed transgenic rescue experiments by crossing these transgenic mice with COL7(m+/-) heterozygous mice. Surprisingly, human COL7 expressed by keratinocytes or by fibroblasts was able to rescue all of the abnormal phenotypic manifestations of the COL7(m-/-) mice, indicating that fibroblasts as well as keratinocytes are potential targets for RDEB gene therapy. Furthermore, we generated transgenic mice with a premature termination codon expressing truncated COL7 protein and performed the same rescue experiments. Notably, the COL7(m-/-) mice rescued with the human COL7A1 allele were able to survive despite demonstrating clinical manifestations very similar to those of human RDEB, indicating that we were able to generate surviving animal models of RDEB with a mutated human COL7A1 gene. This model has great potential for future research into the pathomechanisms of dystrophic epidermolysis bullosa and the development of gene therapies for patients with dystrophic epidermolysis bullosa. (Am J Pathol 2009,175:2508-2517; DOI: 10.2353/ajpath.2009.090347)
• Secondary syphilis mimicking warts in an HIV-positive patient

  S. Shinkuma, R. Abe, M. Nishimura, K. Natsuga, Y. Fujita, T. Nomura, W. Nishie, H. Shimizu

  Sexually Transmitted Infections 85 (6) 484  1368-4973 2009/10 [Refereed][Not invited]
   

  Shinkuma S, Abe R, Nishimura M, Natsuga K, Fujita Y, Nomura T, Nishie W, Shimizu H, Sexually transmitted infections, 2009, vol. 85, no. 6, pp. 484
• A Novel Humanized Neonatal Autoimmune Blistering Skin Disease Model Induced by Maternally Transferred Antibodies

  Wataru Nishie, Daisuke Sawamura, Ken Natsuga, Satoru Shinkuma, Maki Goto, Akihiko Shibaki, Hideyuki Ujiie, Edit Olasz, Kim B. Yancey, Hiroshi Shimizu

  JOURNAL OF IMMUNOLOGY 183 (6) 4088 - 4093 0022-1767 2009/09 [Refereed][Not invited]
   

  All mammal neonates receive maternal Abs for protection against pathogenic organisms in the postnatal environment. However, neonates can experience serious adverse reactions if the Abs transferred from the mother recognize self-molecules as autoAgs. In this study, we describe a novel model for autoimmune disease induced by transferred maternal Abs in genetically transformed Ag-humanized mice progeny. Bullous pemphigoid is the most common life-threatening autoimmune blistering skin disease that affects the elderly, in which circulating IgG autoAbs are directed against epidermal type XVII collagen (COL17). We have established a genetically manipulated experimental mouse model in which maternal Abs against human COL17 are transferred to pups whose skin expresses only human and not mouse COL17, resulting in blistering similar to that seen in patients with bullous pemphigoid. Maternal transfer of pathogenic Abs to humanized neonatal mice is a unique and potential experimental system to establish a novel autoimmune disease model. The Journal of Immunology, 2009, 183: 4088-4093.
• Widespread keratosis follicularis squamosa

  Y. Nomura, M. Abe, K. Natsuga, R. Moriuchi, H. Kawasaki, M. Mayuzumi, A. Yasuoka, H. Shimizu

  CLINICAL AND EXPERIMENTAL DERMATOLOGY 34 (4) 519 - 520 0307-6938 2009/06 [Refereed][Not invited]
   

  Nomura Y, Abe M, Natsuga K, Moriuchi R, Kawasaki H, Mayuzumi M, Yasuoka A, Shimizu H, Clinical and experimental dermatology, 2009, vol. 34, no. 4, pp. 519-520
• Pemphigus foliaceus associated with oesophageal cancer

  S. Shinkuma, M. Akiyama, N. Torii-Saito, K. Natsuga, Y. Tateishi, K. Ito, J. Hirota, Y. Shimizu, T. Shichinohe, H. Shimizu

  JOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY AND VENEREOLOGY 23 (4) 473 - 474 0926-9959 2009/04 [Refereed][Not invited]
   

  Shinkuma S, Akiyama M, Torii-Saito N, Natsuga K, Tateishi Y, Ito K, Hirota J, Shimizu Y, Shichinohe T, Shimizu H, Journal of the European Academy of Dermatology and Venereology : JEADV, 2009, vol. 23, no. 4, pp. 473-474
• Autoantibodies against type XVII collagen C-terminal domain in a patient with bullous pemphigoid associated with psoriasis vulgaris

  D. Inokuma, K. Kodama, K. Natsuga, M. Kasai, M. Abe, W. Nishie, R. Abe, T. Hashimoto, H. Shimizu

  BRITISH JOURNAL OF DERMATOLOGY 160 (2) 451 - 454 0007-0963 2009/02 [Refereed][Not invited]
   

  Inokuma D, Kodama K, Natsuga K, Kasai M, Abe M, Nishie W, Abe R, Hashimoto T, Shimizu H, The British journal of dermatology, 2009, vol. 160, no. 2, pp. 451-454
• Morphological and genetic analysis of steatocystoma multiplex in an Asian family with pachyonychia congenita type 2 harbouring a KRT17 missense mutation

  M. Kanda, K. Natsuga, W. Nishie, M. Akiyama, A. Nagasaki, T. Shimizu, H. Shimizu

  BRITISH JOURNAL OF DERMATOLOGY 160 (2) 465 - 468 0007-0963 2009/02 [Refereed][Not invited]
   

  Kanda M, Natsuga K, Nishie W, Akiyama M, Nagasaki A, Shimizu T, Shimizu H, The British journal of dermatology, 2009, vol. 160, no. 2, pp. 465-468
• Type XVII collagen is a key player in tooth enamel formation.

  Takuya Asaka, Masashi Akiyama, Takanori Domon, Wataru Nishie, Ken Natsuga, Yasuyuki Fujita, Riichiro Abe, Yoshimasa Kitagawa, Hiroshi Shimizu

  The American journal of pathology 174 (1) 91 - 100 0002-9440 2009/01 [Refereed][Not invited]
   

  Inherited tooth enamel hypoplasia occurs due to mutations in genes that encode major enamel components. Enamel hypoplasia also has been reported in junctional epidermolysis bullosa, caused by mutations in the genes that encode type XVII collagen (COL17), a component of the epithelial-mesenchymal junction. To elucidate the pathological mechanisms of the enamel hypoplasia that arise from the deficiency of epithelial-mesenchymal junction molecules, such as COL17, we investigated tooth formation in our recently established Col17(-/-) and Col17 rescued mice. Compared with wild-type mice, the incisors of the Col17(-/-) mice exhibited reduced yellow pigmentation, diminished iron deposition, delayed calcification, and markedly irregular enamel prisms, indicating the presence of enamel hypoplasia. The molars of the Col17(-/-) mice demonstrated advanced occlusal wear. These abnormalities were corrected in the Col17 rescued humanized mice. Thus, the Col17(-/-) mice clearly reproduced the enamel hypoplasia in human patients with junctional epidermolysis bullosa. We were able to investigate tooth formation in the Col17(-/-) mice because the Col17(-/-) genotype is not lethal. Col17(-/-) mouse incisors had poorly differentiated ameloblasts that lacked enamel protein-secreting Tomes' processes and reduced mRNA expression of amelogenin, ameloblastin, and of other enamel genes. These findings indicated that COL17 regulates ameloblast differentiation and is essential for normal formation of Tomes' processes. In conclusion, COL17 deficiency disrupts the epithelial-mesenchymal interactions, leading to both defective ameloblast differentiation and enamel malformation.
• Cutaneous pemphigus vulgaris with skin features similar to the classic mucocutaneous type: a case report and review of the literature

  S. Shinkuma, W. Nishie, A. Shibaki, D. Sawamura, K. Ito, Y. Tsuji-Abe, K. Natsuga, P. T. Chan, M. Amagai, H. Shimizu

  CLINICAL AND EXPERIMENTAL DERMATOLOGY 33 (6) 724 - 728 0307-6938 2008/11 [Refereed][Not invited]
   

  Pemphigus vulgaris (PV) is a life-threatening autoimmune blistering skin disease that specifically involves oral mucosa. It was recently shown that a very small number of patients with PV show no mucous membrane involvement although they have circulating autoantibodies directed against both desmoglein (Dsg)1 and Dsg3 that are associated with histopathological suprabasal acantholysis. These cases are classed as cutaneous-type PV. We report here a case of cutaneous-type PV that occurred in a 50-year-old man. Clinical examination revealed numerous tense and spreading blisters and erosions over the patient's entire body, similar to the classic mucocutaneous-type PV. Interestingly, none of the previously reported patients with cutaneous PV had shown skin features like those of mucocutaneous PV, whereas the present case clearly demonstrated very typical clinical features similar to those in mucocutaneous PV.
• Usefulness of thermography techniques for evaluating the disease activity in Kimura's disease

  S. Shinkuma, W. Nishie, D. Sawamura, K. Natsuga, S. Aoyagi, H. Shimizu

  Clinical and Experimental Dermatology 33 (6) 768 - 771 0307-6938 2008/11 [Refereed][Not invited]
   

  Kimura's disease (KD) is a rare, chronic inflammatory disorder predominantly affecting the head and neck. A case of KD in a 30-year-old man is reported. Thermography was useful for evaluating the activity of the condition. © 2008 The Author(s).
• Acquired perforating dermatosis appearing as elastosis perforans serpiginosa and perforating folliculitis

  R. Abe, S. Murase, Y. Nomura, K. Natsuga, Y. Tateishi, Y. Tomita, Y. Tsuji-Abe, T. Matsumura, H. Shimizu

  CLINICAL AND EXPERIMENTAL DERMATOLOGY 33 (5) 653 - 654 0307-6938 2008/09 [Refereed][Not invited]
   

  Abe R, Murase S, Nomura Y, Natsuga K, Tateishi Y, Tomita Y, Tsuji-Abe Y, Matsumura T, Shimizu H, Clinical and experimental dermatology, 2008, vol. 33, no. 5, pp. 653-654
• Precursor B-cell lymphoblastic lymphoma presented with intraocular involvement and unusual skin manifestations

  Satoru Shinkuma, Ken Natsuga, Masashi Akiyama, Akari Saito, Wataru Saito, Shuichi Ota, Takeshi Kondo, Riichiro Abe, Kazuo Kodama, Hiroshi Shimizu

  ANNALS OF HEMATOLOGY 87 (8) 677 - 679 0939-5555 2008/08 [Refereed][Not invited]
   

  Shinkuma S, Natsuga K, Akiyama M, Saito A, Saito W, Ota S, Kondo T, Abe R, Kodama K, Shimizu H, Annals of hematology, 2008, vol. 87, no. 8, pp. 677-679
• Leukaemic dissemination of Merkel cell carcinoma in a patient with systemic lupus erythematosus

  I. Nemoto, K. C. Sato-Matsumura, Y. Fujita, K. Natsuga, H. Ujiie, Y. Tomita, N. Kato, M. Kondo, K. Ohnishi

  CLINICAL AND EXPERIMENTAL DERMATOLOGY 33 (3) 270 - 272 0307-6938 2008/05 [Refereed][Not invited]
   

  We describe an unusual bone-marrow metastasis of Merkel cell carcinoma (MCC) arising in the right cheek of a 73-year-old woman with systemic lupus erythematosus (SLE) and Sjogren's syndrome, who had been treated with oral prednisolone and methotrexate for 10 years. Seven months after wide local excision followed by local irradiation, the patient presented with thrombocytopaenia. Her bone marrow had been completely replaced by metastatic MCC cells, and metastatic cytokeratin 20-positive cells were also identified in the peripheral blood. To our knowledge, in the English literature, only six cases have been described previously of MCC bone-marrow involvement. Of these six cases, four were immunosuppressed, similar to our case. The high incidence of MCC in immunosuppressed patients such as those with SLE has been discussed previously. We consider that immunosuppression might be associated with bone-marrow metastasis, which is a rare form of MCC.
• Novel ABCA12 mutations identified in two cases of non-bullous congenital ichthyosiform erythroderma associated with multiple skin malignant neoplasia

  Ken Natsuga, Masashi Akiyama, Naoko Kato, Kaori Sakai, Yoriko Sugiyama-Nakagiri, Machiko Nishimura, Hiroo Hata, Masataka Abe, Ken Arita, Yukiko Tsuji-Abe, Takashi Onozuka, Satoru Aoyagi, Kazuo Kodama, Hideyuki Ujiie, Yuki Tomita, Hiroshi Shimizu

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 127 (11) 2669 - 2673 0022-202X 2007/11 [Refereed][Not invited]
   

  Natsuga K, Akiyama M, Kato N, Sakai K, Sugiyama-Nakagiri Y, Nishimura M, Hata H, Abe M, Arita K, Tsuji-Abe Y, Onozuka T, Aoyagi S, Kodama K, Ujiie H, Tomita Y, Shimizu H, The Journal of investigative dermatology, 2007, vol. 127, no. 11, pp. 2669-2673
• Non-Hodgkin lymphoma preceded by recalcitrant eczema

  Ken Natsuga, Riichiro Abe, Hideyuki Ujiie, Akihiko Shibaki, Daisuke Sawamura, Mitsufumi Nishio, Katsuya Fujimoto, Takao Koike, Hiroshi Shimizu

  EUROPEAN JOURNAL OF HAEMATOLOGY 79 (4) 369 - 370 0902-4441 2007/10 [Refereed][Not invited]
   

  Natsuga K, Abe R, Ujiie H, Shibaki A, Sawamura D, Nishio M, Fujimoto K, Koike T, Shimizu H, European journal of haematology, 2007, vol. 79, no. 4, pp. 369-370
• Amicrobal pustulosis asssociated with IgA nephropathy and Sjogren's syndrome

  Ken Natsuga, Daisuke Sawamura, Erina Homma, Toshifumi Nomura, Masataka Abe, Ryuichi Muramatsu, Toshio Mochizuki, Takao Koike, Hiroshi Shimizu

  JOURNAL OF THE AMERICAN ACADEMY OF DERMATOLOGY 3 57 (3) 523 - 526 0190-9622 2007/09 [Refereed][Not invited]
   

  Amicrobial pustulosis is a rare clinical entity characterized by a relapsing pustular eruption, primarily involving the skin folds. We describe a case of amicrobial pustulosis associated with autoimmune diseases (APAD). The patient suffered from IgA nephropathy and Sjogren's syndrome. Skin symptoms were alleviated dramatically after corticosteroid pulse therapy and tonsillectomy.
• Keratoacanthoma developing on nevus sebaceous in a child

  Hideyuki Ujiie, Naoko Kato, Ken Natsuga, Yuki Tomita

  JOURNAL OF THE AMERICAN ACADEMY OF DERMATOLOGY 56 (2) S57 - S58 0190-9622 2007/02 [Refereed][Not invited]
   

  Ujiie H, Kato N, Natsuga K, Tomita Y, Journal of the American Academy of Dermatology, 2007, vol. 56, no. 2 Suppl, pp. S57-8
• Severe cholinergic urticaria successfully treated with scopolamine butylbromide in addition to antihistamines

  H Ujiie, T Shimizu, K Natsuga, K Arita, K Tomizawa, H Shimizu

  CLINICAL AND EXPERIMENTAL DERMATOLOGY 31 (4) 588 - 589 0307-6938 2006/07 [Refereed][Not invited]
   

  Ujiie H, Shimizu T, Natsuga K, Arita K, Tomizawa K, Shimizu H, Clinical and experimental dermatology, 2006, vol. 31, no. 4, pp. 588-589
• Mycosis fungoides bullosa

  K Natsuga, T Shimizu, R Abe, K Kodama, H Shimizu

  ARCHIVES OF DERMATOLOGY 142 (6) 793 - 795 0003-987X 2006/06 [Refereed][Not invited]
   

  Natsuga K, Shimizu T, Abe R, Kodama K, Shimizu H, Archives of dermatology, 2006, vol. 142, no. 6, pp. 793-795
• Two cases of atypical melanocytic lesions in recessive dystrophic epidermolysis bullosa infants

  K Natsuga, M Akiyama, KC Sato-Matsumura, K Tsuchiya, H Shimizu

  CLINICAL AND EXPERIMENTAL DERMATOLOGY 30 (6) 636 - 639 0307-6938 2005/11 [Refereed][Not invited]
   

  Atypical melanocytic lesions (AtML) are known to be associated with epidermolysis bullosa (EB), mainly with the junctional subtype. We report two cases of AtML in two female infants with recessive dystrophic epidermolysis bullosa (RDEB). Both lesions were dark brown- to black-coloured, asymmetric-shaped macules, 3-4 cm in size, with an irregular border and were located on the forearms of two unrelated, 1-year-old female infants. On a clinical and pathological basis, the pigmented macules were diagnosed as AtML in EB patients. There are only a few reports describing in detail the clinical and histopathological features of AtML in RDEB, especially in infant cases. AtML may easily be misdiagnosed as malignant melanoma and, even in infant patients with RDEB, this should be included as one of the differential diagnosis of pigmented lesions.
• Ultrastructural features of trafficking defects are pronounced in melanocytic nevus in Hermansky-Pudlak syndrome type 1

  K Natsuga, M Akiyama, T Shimizu, T Suzuki, S Ito, Y Tomita, J Tanaka, H Shimizu

  JOURNAL OF INVESTIGATIVE DERMATOLOGY 125 (1) 154 - 158 0022-202X 2005/07 [Refereed][Not invited]
   

  Hermansky-Pudiak syndrome (HIRS) is an autosomal recessive disorder characterized by oculocutaneous albinism, a bleeding disorder, and ceroid lipofuscinosis in the lungs and gut. HPS is genetically heterogeneous and the most common variant, HPS type 1, is caused by mutations in HPS1 gene. The protein encoded by HPS1 is considered to facilitate the trafficking of melanocyte-specific gene products into the premelanosome. We report the ultrastructural findings in a melanocytic nevus seen in a 17-y-old Japanese female patient with HPS1 who Is a compound heterozygote of HPS1 mutations, including a novel mutation. Electron microscopy of a pinkish papule corresponding to the melanocytic nevus revealed markedly aberrant, immature melanosomes, large membranous structures, and giant melanosomes in the vicinity of trans-Golgi network, the characteristic abnormalities because of protein trafficking defects in HPS1. These ultrastructural features were far more clearly demonstrated in the nevus cells than in the epidermal melanocytes. Thus, ultrastructural analysis of nevus cells may be an additional diagnostic tool for HPS1 and could give us important clues to further understanding of the pathomechanisms of HPS.

MISC

• 皮膚科にも来た再生医療 皮膚科における自家培養表皮

  夏賀 健  日本臨床皮膚科医会雑誌  41-  (1)  030  -032  2024/01
• 新・皮膚科セミナリウム 水疱症アップデート 表皮水疱症の水疱形成機構

  夏賀 健  日本皮膚科学会雑誌  133-  (12)  2819  -2823  2023/11  

  表皮水疱症は,表皮基底膜領域蛋白の先天的機能不全を原因として,全身の皮膚に水疱やびらんを繰り返す疾患群である.過去30年間の研究によって,ほとんどの原因遺伝子とその水疱形成の機序が解明されてきた.本稿では,古典的4病型とその水疱形成機序ならびに,近年報告された表皮水疱症の新しい原因遺伝子について概説する.(著者抄録)
• 各種難病の最新治療情報 表皮水疱症の最新情報

  夏賀 健  難病と在宅ケア  28-  (10)  49  -51  2023/01
• 重症(旧Herlitz型)接合部型表皮水疱症の1例

  横手 銀珠, 石田 倫子, 工藤 恭子, 市山 正子, 高島 翔太, 夏賀 健  皮膚科の臨床  65-  (1)  69  -73  2023/01  

  ＜文献概要＞日齢1,女児。生下時から左肘と指趾に計2%の皮膚欠損と爪甲の脱落があり,日齢1には口腔粘膜,下顎部,臀部,臍部などにびらんを生じていた。病理組織検査で表皮下に水疱形成がみられ,蛍光抗体直接法でlaminin332が完全欠損しており,重症接合部型表皮水疱症と診断した。その後,全身にびらんが拡大し,体重増加不良,低アルブミン血症,慢性炎症と貧血,脱水があり,適宜アルブミン補充,免疫グロブリン投与や輸血も行ったが,徐々に体重減少を認め,月齢7で死亡した。生下時に軽症であっても,爪異常や特に体幹部に水疱があるときは,重症型の表皮水疱症の可能性も疑い,詳細な診察と組織学的検査による病型分類を行う必要がある。
• 血管型エーラス・ダンロス症候群の遺伝子治療の可能性 siRNA技術を導入した治療法の研究が進められている

  林 周次郎, 夏賀 健  日本医事新報  (5043)  46  -46  2020/12
• 汎発性疣贅症の新知見について 免疫不全を念頭に診断と治療を進める必要がある

  夏賀 健, 新熊 悟  日本医事新報  (5038)  53  -53  2020/11
• 新・皮膚科セミナリウム 自己免疫性水疱症 難治例への対処 自己免疫性表皮下水疱症 診断・病態の最新の進歩

  夏賀 健  日本皮膚科学会雑誌  129-  (13)  2749  -2754  2019/12  [Not refereed][Not invited]
   

  自己免疫性水疱症のうち、表皮基底膜領域蛋白群に対する自己免疫反応による疾患群は自己免疫性表皮下水疱症(類天疱瘡群)と呼称される。患者皮膚検体のみならず、培養細胞や動物実験を用いた実験系によって、近年類天疱瘡群の知識は飛躍的に高まるとともに、治療の選択肢も増えつつある。本稿では、類天疱瘡群の代表的疾患である水疱性類天疱瘡を中心に、診断法の進歩と病態を概説する。(著者抄録)
• 【4疾患からみる膠原病のいま】小児線状強皮症に対してシクロスポリン内服が有効であった2例

  眞井 翔子, 藤田 靖幸, 夏賀 健, 西江 渉, 小野寺 智洋, 岡 敏明, 清水 宏  皮膚科の臨床  61-  (12)  1797  -1802  2019/11  [Not refereed][Not invited]
   

  ＜文献概要＞線状強皮症は皮膚の硬化と萎縮によって,整容面の問題だけではなく不可逆的な筋や関節の拘縮をきたしうる。小児発症が多く,治療効果と副作用の点から治療の選択に苦慮することが多い。われわれは,小児の四肢に生じた線状強皮症2例に対してシクロスポリン内服療法を行ったところ,1例は症状の改善を認め,1例では進行を抑制することができた。ともに経過中に重大な有害事象を生じず,小児のシクロスポリン内服療法は治療の選択肢として有力であると考えた。
• 重症汎発性接合部型表皮水疱症の1家系

  平岡 美樹子, 今門 純久, 夏賀 健, 秋山 真志, 澤村 大輔, 清水 宏  日本皮膚科学会雑誌  129-  (12)  2525  -2532  2019/11  [Not refereed][Not invited]
   

  両親はインド人で従妹婚。第2児と第4児にそれぞれ日齢3日と日齢1日から全身に水疱とびらんが出現。病理検査で表皮下に水疱を認め、免疫染色でlaminin332が完全消失し、重症汎発性接合部型表皮水疱症と診断した。外用処置や抗生剤を投与したが、両児ともにそれぞれ生後1年3ヵ月と1年1ヵ月で死亡した。遺伝子検査では両児ともLAMB3遺伝子のp.Arg569*変異のホモ接合体であった。本変異はこれまで米国、インド、イギリスから報告がある。日本では本症の報告はこれまで15例あるが、本変異の報告はなかった。(著者抄録)
• 治療法の再整理とアップデートのために 専門家による私の治療 鶏眼(うおのめ)・胼胝腫(たこ)

  夏賀 健  日本医事新報  (4976)  48  -48  2019/09  [Not refereed][Not invited]
  
• 【内科医のための皮膚疾患アトラス-日常診療における部位別皮疹の診かた-】(Part2)症例 皮疹(皮膚所見)からの診断と治療へのアプローチ<部位別> 全身 全身の水ぶくれと紅斑

  夏賀 健  診断と治療  107-  (Suppl.)  322  -323  2019/03
• 【内科医のための皮膚疾患アトラス-日常診療における部位別皮疹の診かた-】(Part2)症例 皮疹(皮膚所見)からの診断と治療へのアプローチ<部位別> 全身 全身の水ぶくれ

  夏賀 健  診断と治療  107-  (Suppl.)  324  -325  2019/03
• 皮膚加齢における基底膜蛋白の生理学的意義の解明

  夏賀 健  コスメトロジー研究報告  26-  94  -97  2018/09  [Not refereed][Not invited]
  
• トラニラストが奏効したannular elastolytic giant cell granulomaの1例

  藤村 悠, 夏賀 健, 伊東 孝政, 青柳 哲, 清水 宏  臨床皮膚科  72-  (9)  681  -684  2018/08  [Not refereed][Not invited]
   

  ＜文献概要＞67歳,男性.約10年前から両前腕に環状の紅斑を自覚していた.半年前に近医内科で外用剤を処方されたが,改善しないため当科を受診した.両前腕に,径5cm大までの楕円形から不整形,辺縁の色調が強い淡紅褐色局面を数個認めた.病理組織像では,真皮上層に肉芽腫の形成と多核巨細胞による弾力線維の貪食像を認めた.以上からannular elastolytic giant cell granulomaと診断した.トラニラスト内服で治療を開始したところ,皮疹は改善傾向を示した.本疾患に対するトラニラストの奏効例は徐々に報告されてきており,試みる価値のある治療である.
• 表皮下水疱症 発症機序/新規治療法の最新の進歩

  夏賀 健  日本皮膚科学会雑誌  128-  (5)  971  -971  2018/05  [Not refereed][Not invited]
  
• 17型コラーゲンは加齢による表皮の過増殖を抑制する

  渡邉 美佳, 夏賀 健, 西江 渉, 藤村 悠, 氏家 英之, 尾崎 倫孝, 清水 宏  日本皮膚科学会雑誌  128-  (4)  607  -607  2018/04  [Not refereed][Not invited]
  
• 陰圧閉鎖療法が奏功したHydroxyurea(HU)による下腿潰瘍の2例

  小住 英之, 渡邉 美佳, 夏賀 健, 山口 泰之, 須貝 達朗, 中山 ちひろ, 中里 信一, 新熊 悟, 清水 宏  日本皮膚科学会雑誌  128-  (4)  607  -607  2018/04  [Not refereed][Not invited]
  
• 脾摘後に自然消退した続発性皮膚B細胞リンパ腫の1例

  眞井 翔子, 夏賀 健, 清水 宏, 白鳥 聡一, 高橋 祥公  日本皮膚科学会雑誌  128-  (4)  608  -608  2018/04  [Not refereed][Not invited]
  
• BNIP3は紫外線から皮膚を保護する際に重要な役割を果たす

  森田 遼, 森山 麻里子, 森田 貴士, 丸谷 祐樹, 後藤 ありさ, 松本 諭以子, 夏賀 健, 早川 堯夫, 森山 博由  日本薬学会年会要旨集  138年会-  (3)  133  -133  2018/03  [Not refereed][Not invited]
  
• BNIP3は紫外線から皮膚を保護する際に重要な役割を果たす

  森田遼, 森山麻里子, 森田貴士, 丸谷祐樹, 後藤ありさ, 松本諭以子, 夏賀健, 早川堯夫, 森山博由  日本薬学会年会要旨集(CD-ROM)  138th-  (3)  ROMBUNNO.26PA‐am169S  -133  2018/03  [Not refereed][Not invited]
  
• 小児線状強皮症に対してシクロスポリン内服が有効であった3例

  眞井 翔子, 藤田 靖幸, 宮内 俊成, 藤村 悠, 夏賀 健, 乃村 俊史, 西江 渉, 清水 宏, 小野寺 智洋, 岡 敏明  日本皮膚科学会雑誌  128-  (1)  62  -62  2018/01  [Not refereed][Not invited]
  
• Bnip3が誘導するオートファジーは紫外線ストレスから表皮を保護する

  丸谷 祐樹, 森山 麻里子, 森田 貴士, 後藤 ありさ, 森田 遼, 松本 諭以子, 夏賀 健, 早川 堯夫, 森山 博由  生命科学系学会合同年次大会  2017年度-  [1P  -0561]  2017/12  [Not refereed][Not invited]
  
• 【ヒト化マウスモデル】 自己抗原のヒト化による自己免疫性表皮下水疱症モデル

  夏賀 健, 清水 宏  臨床免疫・アレルギー科  68-  (3)  271  -277  2017/09  [Not refereed][Not invited]
  
• 皮膚基底膜構築における17型コラーゲン細胞外領域切断の生理機能の解析

  西村 真智子, 西江 渉, 新熊 悟, 夏賀 健, 中村 秀樹, 清水 宏, 白藤 宜紀, 岩月 啓氏, 澤村 大輔  日本皮膚科学会雑誌  127-  (9)  2111  -2111  2017/08  [Not refereed][Not invited]
  
• Matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF/MS)が診断に有用であったMicrosporum canis感染症の家族例

  山口 泰之, 藤田 靖幸, 白戸 貴久, 新熊 悟, 夏賀 健, 乃村 俊史, 清水 宏, 福元 達也  日本皮膚科学会雑誌  127-  (9)  2115  -2115  2017/08  [Not refereed][Not invited]
  
• 単回使用陰圧治療システムが有効であった壊疽性膿皮症の2例

  山口 泰之, 柳 輝希, 佐藤 一正, 葭本 倫大, 平田 悠, 氏家 韻欣, 西村 真智子, 夏賀 健, 清水 宏, 椎谷 千尋, 月永 一郎, 松村 哲理  日本皮膚科学会雑誌  127-  (9)  2118  -2118  2017/08  [Not refereed][Not invited]
  
• bcl2ファミリー分子BNIP3はオートファジーを介した表皮における形態維持を担う

  丸谷 祐樹, 森山 麻里子, 森田 貴士, 中島 佑香, 後藤 ありさ, 森田 遼, 夏賀 健, 早川 堯夫, 森山 博由  日本薬学会年会要旨集  137年会-  (3)  152  -152  2017/03  [Not refereed][Not invited]
  
• 【新生児の皮膚病】 (コラム1)出生前診断について

  夏賀 健  Visual Dermatology  16-  (3)  276  -277  2017/02  [Not refereed][Not invited]
  
• 広範囲に皮疹を生じた長島型掌蹠角化症の1例

  宮内 俊成, 乃村 俊史, 鈴木 翔多朗, 大口 由香, 山口 泰之, 新熊 悟, 夏賀 健, 藤田 靖幸, 清水 宏  日本皮膚科学会雑誌  126-  (9)  1725  -1725  2016/08  [Not refereed][Not invited]
  
• 優性栄養障害型表皮水疱症(DDEB)患者に生じた後天性表皮水疱症(EBA)の1例

  林 良太, 伊藤 明子, 増井 由紀子, 伊藤 雅章, 阿部 理一郎, 下村 裕, 夏賀 健, 渡邉 美佳, 岩田 浩明, 新熊 悟  日本皮膚科学会雑誌  126-  (7)  1320  -1320  2016/06  [Not refereed][Not invited]
  
• Herlitz型接合部型表皮水疱症の1例

  横山 華英, 木村 有太子, 高森 健二, 須賀 康, 夏賀 健, 西江 渉, 清水 宏  日本皮膚科学会雑誌  126-  (7)  1321  -1321  2016/06  [Not refereed][Not invited]
  
• 病理学的に乾癬様の変化を呈した菌状息肉症の2例

  山口 泰之, 藤田 靖幸, 平田 悠, 西村 真智子, 夏賀 健, 清水 宏, 肥田 時征, 加藤 直子  日本皮膚科学会雑誌  126-  (7)  1331  -1331  2016/06  [Not refereed][Not invited]
  
• アトピー性皮膚炎/魚鱗癬における細菌叢と抗菌ペプチド

  夏賀 健  日本皮膚科学会雑誌  126-  (5)  793  -793  2016/05  [Not refereed][Not invited]
  
• 表皮細胞の分化・増殖における基底膜の役割

  夏賀 健  日本皮膚科学会雑誌  126-  (5)  841  -841  2016/05  [Not refereed][Not invited]
  
• 広範囲に皮疹を生じた長島型掌蹠角化症の1例

  宮内 俊成, 乃村 俊史, 鈴木 翔多朗, 大口 由香, 山口 泰之, 新熊 悟, 夏賀 健, 藤田 靖幸, 清水 宏  日本皮膚科学会雑誌  126-  (5)  979  -979  2016/05  [Not refereed][Not invited]
  
• 長島型掌蹠角化症の1例

  宮内 俊成, 乃村 俊史, 鈴木 翔多朗, 山口 泰之, 夏賀 健, 藤田 靖幸, 清水 宏  角化症研究会記録集  30-  78  -82  2016/03  [Not refereed][Not invited]
  
• bc12ファミリー分子BNIP3はオートファジーを介して表皮の分化および形態維持を行う

  久保 嘉一, 森山 麻里子, 中島 佑香, 後藤 ありさ, 夏賀 健, 早川 堯夫, 森山 博由  日本薬学会年会要旨集  136年会-  (3)  146  -146  2016/03  [Not refereed][Not invited]
  
• 病理学的に乾癬様の変化を呈した菌状息肉症の2例

  山口 泰之, 藤田 靖幸, 平田 悠, 西村 真智子, 夏賀 健, 清水 宏, 肥田 時征, 加藤 直子  日本皮膚科学会雑誌  126-  (2)  167  -167  2016/02  [Not refereed][Not invited]
  
• 進行性顔面片側萎縮症の1例

  堀田 萌子, 藤田 靖幸, 葭本 倫大, 剱持 靖子, 夏賀 健, 冨澤 幸生, 清水 宏  皮膚科の臨床  58-  (1)  150  -151  2016/01  [Not refereed][Not invited]
   

  9歳男児。約半年前から左頬部に陥凹が出現し、徐々に拡大してきたため、当科を紹介受診した。初診時、左頬全体に常色で境界不明瞭な陥凹を認め、病理組織学的所見・MRI所見より、進行性顔面片側萎縮症と診断した。診断後は無治療にて経過観察しているが、現在までに症状の増悪はみられない。
• 水疱性類天疱瘡におけるIgE自己抗体の解析

  守内 玲寧, 西江 渉, 氏家 英之, 夏賀 健, 清水 宏  日本皮膚科学会雑誌  125-  (12)  2301  -2301  2015/11  [Not refereed][Not invited]
  
• Piezogenic pedal papulesの3例

  椎谷 千尋, 夏賀 健, 泉 健太郎, 青柳 哲, 清水 宏  臨床皮膚科  69-  (10)  731  -734  2015/09  [Not refereed][Not invited]
   

  症例1:28歳,女性.主訴は両踵部の有痛性丘疹.初診の2年前から,両踵部の丘疹を自覚した.丘疹の数が増加し,疼痛を伴うようになり,受診した.立位時,両踵部内側に7〜8mm大で正常皮膚色の軟らかい丘疹を10個程度認めた.座位で丘疹は消失した.症例2:19歳,男性.主訴は両踵部の有痛性丘疹.数年前から両踵部に疼痛を自覚していた.足底の圧迫により出現する,両踵部内側に正常皮膚色の丘疹を10個程度認めた.圧迫解除によって皮疹は消失した.症例3:46歳,女性.主訴は両踵部の有痛性丘疹.20歳台から両踵部に皮疹を自覚していた.疼痛を伴うようになり,受診した.両踵部内側に正常皮膚色の丘疹が散在していた.皮疹は立位で誘発され,座位で消失した.いずれの症例もpiezogenic pedal papulesと診断し,経過観察とした.本疾患は日常的に遭遇するものであり,無用な侵襲的検査を避けるためにも,疾患を広く周知することが重要である.(著者抄録)
• Autoantibodies to parts of type XVII collagen outside of the non-collageneous 16A domain lead to mild bullous pemphigoid due to the non-depletion of autoantigen

  H. Iwata, K. Imafuku, K. Izumi, M. Wada, K. Natsuga, H. Ujiie, W. Nishie, H. Shimizu  JOURNAL OF INVESTIGATIVE DERMATOLOGY  135-  S4  -S4  2015/09  [Not refereed][Not invited]
  
• 角化症と水疱症 その基礎と臨床 水疱症update 2014

  夏賀 健  西日本皮膚科  77-  (3)  281  -281  2015/06  [Not refereed][Not invited]
  
• 【骨格筋症候群(第2版)-その他の神経筋疾患を含めて-】[上] 筋ジストロフィーおよび膜イオンチャネル異常症 先天性筋ジストロフィー(CMD) 非福山型先天性筋ジストロフィー 表皮水疱症 筋ジストロフィー症候群(プレクチン欠損症)

  夏賀 健  日本臨床  別冊-  (骨格筋症候群(上))  168  -172  2015/05  [Not refereed][Not invited]
  
• 進行性顔面片側萎縮症の1例

  堀田 萌子, 藤田 靖幸, 葭本 倫大, 剱持 靖子, 夏賀 健, 清水 宏, 冨澤 幸生  日本皮膚科学会雑誌  125-  (6)  1268  -1269  2015/05  [Not refereed][Not invited]
  
• Piezogenic pedal papulesの3例

  椎谷 千尋, 夏賀 健, 泉 健太郎, 青柳 哲, 清水 宏  日本皮膚科学会雑誌  125-  (6)  1281  -1281  2015/05  [Not refereed][Not invited]
  
• 抗NC16A抗体を有さない軽症の水疱性類天疱瘡の1例

  今福 恵輔, 岩田 浩明, 泉 健太郎, 夏賀 健, 氏家 英之, 西江 渉, 清水 宏  日本皮膚科学会雑誌  125-  (4)  915  -915  2015/04  [Not refereed][Not invited]
  
• 水疱性類天疱瘡自己抗原である17型コラーゲンの異なるエピトープに対する抗体の病原性解析

  和田 麻友美, 西江 渉, 氏家 英之, 泉 健太郎, 岩田 浩明, 夏賀 健, 清水 宏, 北川 善政  北海道歯学雑誌  35-  (2)  188  -188  2015/03  [Not refereed][Not invited]
  
• 【臨床医学の展望-30領域のトピックス】 皮膚科学

  清水 宏, 夏賀 健  日本医事新報  (4740)  74  -77  2015/02  [Not refereed][Not invited]
  
• 進行性顔面片側萎縮症の1例

  堀田 萌子, 藤田 靖幸, 葭本 倫大, 剱持 靖子, 夏賀 健, 冨澤 幸生, 清水 宏  日本皮膚科学会雑誌  125-  (1)  104  -104  2015/01  [Not refereed][Not invited]
  
• 結節性強皮症の1例

  中里 信一, 乃村 俊史, 山根 尚子, 新熊 悟, 夏賀 健, 藤田 靖幸, 有田 賢, 竹中 ちひろ, 清水 宏  日本皮膚科学会雑誌  125-  (1)  104  -104  2015/01  [Not refereed][Not invited]
  
• 中年女性の前腕に生じたelastotic bandsの1例

  羽賀 直哉, 阿部 理一郎, 森田 裕介, 藤村 悠, 夏賀 健, 乃村 俊史, 熊切 正信, 清水 宏  日本皮膚科学会雑誌  124-  (14)  3184  -3184  2014/12  [Not refereed][Not invited]
  
• 鎖骨内金属インプラントによる皮膚潰瘍の1例

  北村 真也, 夏賀 健, 今福 恵輔, 本間 英里奈, 山根 尚子, 青柳 哲, 清水 宏, 松村 哲理  日本皮膚科学会雑誌  124-  (14)  3190  -3190  2014/12  [Not refereed][Not invited]
  
• 50歳代女の前腕に生じたelastotic bandsの1例

  羽賀 直哉, 阿部 理一郎, 森田 裕介, 藤村 悠, 夏賀 健, 乃村 俊史, 清水 宏, 熊切 正信  日本皮膚科学会雑誌  124-  (11)  2134  -2134  2014/10  [Not refereed][Not invited]
  
• Piezogenic pedal papulesの3例

  椎谷 千尋, 夏賀 健, 泉 健太郎, 青柳 哲, 清水 宏  日本皮膚科学会雑誌  124-  (11)  2134  -2134  2014/10  [Not refereed][Not invited]
  
• METAL IMPLANT-INDUCED SKIN ULCER MIMICKING SCROFULODERMA

  Shinya Kitamura, Ken Natsuga, Keisuke Imafuku, Erina Homma, Naoko Yamane, Satoru Aoyagi, Tetsuri Matsumura, Hiroshi Shimizu  JOURNAL OF DERMATOLOGY  41-  6  -6  2014/10  [Not refereed][Not invited]
  
• 【水疱症の治療最前線】 類天疱瘡・粘膜類天疱瘡の治療

  夏賀 健, 西江 渉  Derma.  (222)  15  -19  2014/09  [Not refereed][Not invited]
   

  類天疱瘡群は、表皮基底膜構成蛋白に対する自己抗体を背景に、表皮下水疱を生じる自己免疫性水疱症である。類天疱瘡群は、臨床像と病理像、蛍光抗体法、ELISA法、免疫ブロット法などの組み合わせによって診断される。治療法の根幹となるのは中等量のステロイドの全身投与である。通常プレドニゾロン換算で0.5mg/kg/dayのステロイド投与で加療する。軽症例では、strongestクラスのステロイド外用剤のみで加療することが可能なこともある。重症例や難治例では、プレドニゾロン換算で1mg/kg/day、ステロイドパルス療法、血漿交換、免疫抑制剤の併用、免疫グロブリン大量静注などを組み合わせる。それ以外の症例は、プレドニゾロン換算で通常0.5mg/kg/dayのステロイド投与で加療する。抗CD20モノクローナル抗体や、抗IgEモノクローナル抗体の有効性が海外で報告されつつあり、本邦でも将来的に使用できる可能性がある。(著者抄録)
• 膿疱性乾癬と抗ラミニンγ1類天疱瘡を合併した1例

  藤村 悠, 夏賀 健, 濱出 洋平, 野村 友希子, 清水 宏, 加来 洋, 村松 隆一  日本皮膚科学会雑誌  124-  (10)  1936  -1936  2014/09  [Not refereed][Not invited]
  
• 鎖骨内金属インプラントによる皮膚潰瘍の1例

  北村 真也, 夏賀 健, 今福 恵輔, 本間 英里奈, 山根 尚子, 青柳 哲, 清水 宏, 松村 哲理  日本皮膚科学会雑誌  124-  (10)  1941  -1941  2014/09  [Not refereed][Not invited]
  
• 血清中に抗デスモコリン3IgG抗体を検出した増殖性天疱瘡の1例

  守内 玲寧, 菊地 一博, 伊東 孝政, 清水 聡子, 村松 隆一, 夏賀 健, 清水 宏  日本皮膚科学会雑誌  124-  (10)  1942  -1942  2014/09  [Not refereed][Not invited]
  
• 抗デスモグレイン自己抗体を認めた口腔扁平上苔癬の2例

  村松 憲, 西江 渉, 夏賀 健, 藤田 靖幸, 岩田 浩明, 清水 宏, 山田 珠希, 山下 映美, 浅香 卓哉  日本皮膚科学会雑誌  124-  (10)  1946  -1946  2014/09  [Not refereed][Not invited]
  
• Plasmin but not neutrophil elastase nor MMP9 primarily cleaves the juxtamembranous NC16A domain of collagen XVII

  W. Nishie, W. Mayumi, K. Natsuga, K. Izumi, H. Shimizu  JOURNAL OF INVESTIGATIVE DERMATOLOGY  134-  S13  -S13  2014/09  [Not refereed][Not invited]
  
• トラニラストが奏効したannular elastolytic giant cell granulomaの1例

  藤村 悠, 夏賀 健, 伊東 孝政, 青柳 哲, 清水 宏  日本皮膚科学会雑誌  124-  (5)  984  -984  2014/04  [Not refereed][Not invited]
  
• 形質細胞性口唇炎と日光口唇炎のダーモスコピー所見の検討

  伊東 孝政, 夏賀 健, 谷村 心太郎, 青柳 哲, 清水 宏  日本皮膚科学会雑誌  124-  (4)  775  -775  2014/04  [Not refereed][Not invited]
  
• ケラチン9遺伝子変異を認めたepidermolytic palmoplantar keratodermaの1例

  高島 翔太, 乃村 俊史, 鈴木 翔多朗, 藤村 悠, 中里 信一, 夏賀 健, 佐藤 英嗣, 阿部 理一郎, 清水 宏  日本皮膚科学会雑誌  124-  (4)  792  -792  2014/04  [Not refereed][Not invited]
  
• 自己抗原の枯渇を標的とした水疱性類天疱瘡の新規治療法開発

  夏賀 健  日本皮膚科学会雑誌  124-  (4)  866  -866  2014/04  [Not refereed][Not invited]
  
• The role of decreased skin tension in the pathogenesis of bullous pemphigoid: a newly proposed hypothesis

  Hans I-Chen Harn, Chao-Kai Hsu, Michael Hughes, Ken Natsuga, Hiroshi Shimizu, Julia Yu-Yun Lee, Ming-Jer Tang  JOURNAL OF INVESTIGATIVE DERMATOLOGY  134-  (4)  1178  -1178  2014/04  [Not refereed][Not invited]
  
• Pathogenetic mechanisms in autoimmune skin fragility

  Ken Natsuga  JOURNAL OF INVESTIGATIVE DERMATOLOGY  134-  (4)  1171  -1171  2014/04  [Not refereed][Not invited]
  
• 肛門周囲に限局した水疱性類天疱瘡の1例

  山本 洋平, 阿部 理一郎, 中里 信一, 夏賀 健, 乃村 俊史, 西江 渉, 清水 宏  日本皮膚科学会雑誌  123-  (8)  1548  -1548  2013/07  [Not refereed][Not invited]
  
• 形質細胞性口唇炎と日光口唇炎のダーモスコピー所見の検討

  伊東 孝政, 夏賀 健, 谷村 心太郎, 青柳 哲, 清水 宏  日本皮膚科学会雑誌  123-  (8)  1548  -1548  2013/07  [Not refereed][Not invited]
  
• トラニラストが奏効したannular elastolytic giant cell granulomaの1例

  藤村 悠, 夏賀 健, 伊東 孝政, 青柳 哲, 清水 宏  日本皮膚科学会雑誌  123-  (8)  1550  -1550  2013/07  [Not refereed][Not invited]
  
• Skin barrier defect allows more abundant microbiota in skin and induces skin inflammation and the expression of antimicrobial peptides independently of microbiota

  K. Natsuga, S. Cipolat, F. M. Watt  JOURNAL OF INVESTIGATIVE DERMATOLOGY  133-  S192  -S192  2013/05  [Not refereed][Not invited]
  
• Essential role of collagen XVII in basement membrane formation and keratinocyte migration

  M. Nishimura, W. Nishie, Y. Shirafuji, S. Shinkuma, K. Natsuga, H. Nakamura, D. Sawamura, K. Iwatsuki, H. Shimizu  JOURNAL OF INVESTIGATIVE DERMATOLOGY  133-  S151  -S151  2013/05  [Not refereed][Not invited]
  
• アトピー性皮膚炎および先天性表皮水疱症として経過観察されていたKindler症候群の2例

  神谷 浩二, 白藤 宜紀, 青山 裕美, 岩月 啓氏, 新熊 悟, 夏賀 健, 中村 秀樹, 西江 渉, 清水 宏  日本皮膚科学会雑誌  123-  (4)  456  -456  2013/04  [Not refereed][Not invited]
  
• ケラチン5のコイル1A領域N末端部にミスセンス変異を生じた単純型表皮水疱症

  新熊 悟, 西江 渉, 夏賀 健, 氏家 英之, 中村 秀樹, 清水 宏, Jacyk Witold, 秋山 真志  日本皮膚科学会雑誌  123-  (4)  461  -461  2013/04  [Not refereed][Not invited]
  
• Exophiala speciesによるPhaeohyphomycosisの1例

  宮内 俊成, 阿部 理一郎, 柴 景子, 村田 純子, 山根 尚子, 夏賀 健, 亀井 克彦, 清水 宏  日本皮膚科学会雑誌  123-  (5)  989  -989  2013/04  [Not refereed][Not invited]
  
• 毛包系腫瘍の多発を認めたKID症候群の1例

  小田 裕次郎, 古結 英樹, 成田 幸代, 瀬戸山 充, 神田 真聡, 夏賀 健, 秋山 真志  角化症研究会記録集  27-  68  -71  2013/03  [Not refereed][Not invited]
  
• Antibodies to pathogenic epitopes on type X VII collagen cause skin fragility in a complement-dependent and -independent manner

  夏賀 健, 西江 渉, 新熊 悟, 氏家 英之, 西村 真知子, 澤村 大輔, 清水 宏  北海道醫學雜誌 = Acta medica Hokkaidonensia  87-  (6)  262  -262  2012/11/01
• Chemical Carcinogenesis in Mice With a Defective Epidermal Barrier - Exploring the Connection Between Skin Barrier and Cancer Susceptibility

  S. Cipolat, K. Natsuga, L. M. Sevilla, R. Nachat, F. M. Watt  EUROPEAN JOURNAL OF CANCER  48-  S170  -S170  2012/07  [Not refereed][Not invited]
  
• 肛囲に限局したlinear IgA/IgG bullous dermatosisの1例

  清水 聡子, 夏賀 健, 新熊 悟, 安居 千賀子, 土屋 喜久夫, 清水 宏  日本皮膚科学会雑誌  122-  (4)  1210  -1210  2012/04  [Not refereed][Not invited]
  
• 抗BP230抗体陽性pemphigoid nodularisの1例

  新熊 悟, 猪熊 大輔, 夏賀 健, 氏家 英之, 西江 渉, 清水 宏  日本皮膚科学会雑誌  122-  (3)  625  -625  2012/03  [Not refereed][Not invited]
  
• 肛囲に限局したlinear IgA/IgG bullous dermatosisの1例

  清水 聡子, 安居 千賀子, 林 韻欣, 土屋 喜久夫, 夏賀 健, 新熊 悟, 清水 宏  日本皮膚科学会雑誌  122-  (2)  379  -379  2012/02  [Not refereed][Not invited]
  
• 常染色体劣性乏毛症患者に高頻度に認められたLIPH founder変異とその機能解析

  新熊 悟, 秋山 真志, 夏賀 健, 乃村 俊史, 有田 賢, 西江 渉, 清水 宏, 井上 飛鳥, 青木 淳賢  日本皮膚科学会雑誌  122-  (2)  379  -379  2012/02  [Not refereed][Not invited]
  
• 劣性栄養障害型表皮水疱症患者における遺伝子型-表現型の相互関係の解析

  新熊 悟, 西江 渉, 夏賀 健, 伊藤 圭, 中村 秀樹, 清水 宏  日本皮膚科学会雑誌  122-  (2)  381  -381  2012/02  [Not refereed][Not invited]
  
• 日本人に生じたDuhring疱疹状皮膚炎における表皮トランスグルタミナーゼ(eTG)抗体価の検討

  新熊 悟, 阿部 理一郎, 林 欣宇, 夏賀 健, 氏家 英之, 西江 渉, 清水 宏, 大田 光仁, 土屋 喜久夫, 福本 隆也, 浅田 秀夫  日本皮膚科学会雑誌  122-  (2)  383  -383  2012/02  [Not refereed][Not invited]
  
• 最近経験した先天性表皮水疱症の2例

  木村 亜矢子, 神戸 直智, 松江 弘之, 夏賀 健, 清水 宏  日本皮膚科学会雑誌  122-  (2)  435  -435  2012/02  [Not refereed][Not invited]
  
• 【橋本公二コネクション-薬疹・水疱症・そして皮膚科医人生-】 (Part2)水疱症関連 (case 07)非Herlitz型接合部型表皮水疱症

  松本 圭子, 橋本 公二, 薬師寺 直喜, 西江 渉, 夏賀 健, 中村 秀樹, 清水 宏  Visual Dermatology  11-  (2)  165  -167  2012/01  [Not refereed][Not invited]
  
• 複数の表皮基底膜構成蛋白に対する自己抗体を認めた自己免疫性水疱症の1例

  菊地 一博, 佐藤 英嗣, 夏賀 健, 梶田 哲  日本皮膚科学会雑誌  122-  (1)  54  -54  2012/01  [Not refereed][Not invited]
  
• プレクチンの発現様式の違いによって2種類の表皮水疱症の臨床型が決定される

  夏賀 健, 西江 渉, 秋山 志, 新熊 悟, 中村 秀樹, 長崎 暁理, 澤村 大輔, 清水 宏, Has Cristina, Bruckner-Tuderman Leena, 大内 健嗣, 石河 晃, 平子 善章, 尾張部 克志  日本皮膚科学会雑誌  122-  (1)  57  -57  2012/01  [Not refereed][Not invited]
  
• 担癌患者に生じた自己免疫性水疱症の1例

  守内 玲寧, 阿部 理一郎, 夏賀 健, 乃村 俊史, 清水 宏, 三田村 卓, 櫻木 範明, 橋本 隆  日本皮膚科学会雑誌  121-  (14)  3355  -3355  2011/12  [Not refereed][Not invited]
  
• 先天性爪甲硬厚症2型におけるKRT17遺伝子変異検索

  神田 真聡, 夏賀 健, 西江 渉, 秋山 真志, 長崎 暁理, 清水 宏, 清水 忠道  日本皮膚科学会雑誌  121-  (14)  3357  -3357  2011/12  [Not refereed][Not invited]
  
• 自己免疫性表皮下水疱症の診断

  夏賀 健  札幌社会保険総合病院医誌  20-  (2)  48  -52  2011/12  [Not refereed][Not invited]
  
• 先天性表皮水疱症の1例

  澤井 孝宏, 川見 健也, 清原 隆宏, 熊切 正信, 夏賀 健, 谷岡 未樹, 清水 宏  日本皮膚科学会雑誌  121-  (5)  906  -906  2011/04  [Not refereed][Not invited]
  
• 先天性魚鱗癬様紅皮症、葉状魚鱗癬の病因 ABCA12遺伝子変異とTGM1遺伝子変異

  坂井 香織, 秋山 真志, 柳 輝希, 夏賀 健, 清水 宏  角化症研究会記録集  25-  52  -54  2011/03  [Not refereed][Not invited]
  
• 【新入医局員最初の一歩 目で見てわかる皮膚の科学】 (Part2)ミクロ検査の実際 遺伝子検査の実際

  夏賀 健, 秋山 真志  Visual Dermatology  10-  (4)  394  -397  2011/03  [Not refereed][Not invited]
  
• VII型コラーゲンとラミニン332に対する自己抗体を認めた小児表皮下水疱症の1例

  林 欣宇, 柳 輝希, 秋山 真志, 飯谷 麻里, 守内 玲寧, 夏賀 健, 新熊 悟, 猪熊 大輔, 有田 賢, 清水 宏  日本皮膚科学会雑誌  121-  (3)  585  -585  2011/03  [Not refereed][Not invited]
  
• クロベタゾールプロピオン酸エステル軟膏外用が奏効した高齢者水疱性類天疱瘡の一例

  新熊 悟, 松村 和子, 夏賀 健  札幌社会保険総合病院医誌  19-  (2)  53  -57  2010/12  [Not refereed][Not invited]
   

  78歳男。初診1ヵ月前から掌蹠を除くほぼ全身に浮腫性紅斑が散在、一部では多発、癒合し、水疱、びらんを伴い、粘膜病変は認めないが、両下腿に著明な浮腫を認めた。白血球数は上昇、Hb、血小板数は低値、BUN、CREおよびCRPは上昇していた。水疱を含む浮腫性紅斑の生検では、表皮下に裂隙があり、裂隙内と真皮上層の血管周囲にはリンパ球と組織球および好酸球が浸潤していた。ELISA indexはBP180 ELISA index 18、BP230 ELISA index 1.9(cut off)、免疫ブロット法では、IgGが230kDa、180kDaともに陰性で、BP180NC16部位リコンビナント蛋白を用いた免疫ブロット法でもIgG陰性であった。BP180c末領域のリコンビナント蛋白に対しIgG陽性を示した。以上より、BP180c未領域に対する自己抗体を有する水疱性類天疱瘡(BP)と診断した。腎不全の併発のためクロベタゾールプロピオン酸エステル0.05%軟膏の40g/day外用を14日間行ったところ、皮疹は著明に改善しステロイド外用減量後も再燃は認めていない。
• アトピー性皮膚炎との鑑別を要したKindler症候群の一例

  白藤 宜紀, 青山 裕美, 岩月 啓氏, 梅村 茂夫, 新熊 悟, 夏賀 健, 中村 秀樹, 西江 渉, 清水 宏  Journal of Environmental Dermatology and Cutaneous Allergology  4-  (5)  437  -437  2010/11  [Not refereed][Not invited]
  
• Erratum: Bone marrow transplantation restores epidermal basement membrane protein expression and rescues epidermolysis bullosa model mice (Proceedings of the National Academy of Sciences of the United States of America (2010) 107, (14345-14350) DOI: 10.10

  Yasuyuki Fujita, Riichiro Abe, Daisuke Inokuma, Mikako Sasaki, Daichi Hoshina, Ken Natsuga, Wataru Nishie, James R. McMillan, Hideki Nakamura, Tadamichi Shimizu, Masashi Akiyama, Daisuke Sawamura, Hiroshi Shimizu  Proceedings of the National Academy of Sciences of the United States of America  107-  14514  2010/08/10  [Not refereed][Not invited]
  
• 複数の表皮基底膜構成蛋白に対する自己抗体を認めた自己免疫性水疱症の1例

  菊地 一博, 佐藤 英嗣, 夏賀 健, 梶田 哲  西日本皮膚科  72-  (3)  300  -300  2010/06  [Not refereed][Not invited]
  
• 筋ジストロフィー型単純型表皮水疱症の1例

  大内 健嗣, 舩越 建, 谷川 瑛子, 小堺 有史, 夏賀 健, 秋山 真志, 石河 晃  日本小児皮膚科学会雑誌  29-  (1)  43  -48  2010/05  [Not refereed][Not invited]
   

  24歳女。患者は足底および足背の水疱を主訴とした。乳児期から外力が加わる四肢を中心に水疱や糜爛を形成し、これが瘢痕を残さず上皮化することを繰返していた。更に23歳時には筋ジストロフィーの疑いで経過観察となったが、皮疹は四肢末端に限局しており、病理組織学的所見では表皮直下に裂隙の形成がみられた。そして、裂隙内にはフィブリン様物質が析出していた。更に真皮浅層の血管周囲性には軽度のリンパ球浸潤が認められ、電子顕微鏡所見では基底板上にヘミデスモゾームを残し、ヘミデスモゾーム直上の基底細胞内で表皮内水疱が形成されていた。以上より、本症例はプレクチン異常が疑われ、免疫組織染色を施行したところ、プレクチン分子の特異的欠損が証明され、筋ジストロフィーを伴う単純型表皮水疱症と診断された。
• 先天性表皮水疱症の遺伝カウンセリング

  笠井 靖代, 渡邊 理子, 有馬 香織, 今井 庸子, 窪田 祥吾, 杉本 充弘, 夏賀 健, 清水 宏  日本遺伝カウンセリング学会誌  31-  (1)  62  -62  2010/04  [Not refereed][Not invited]
  
• 高頻度に認められるLIPH founder変異を有した常染色体劣性乏毛症の4家系

  新熊 悟, 秋山 真志, 夏賀 健, 乃村 俊史, 有田 賢, 西江 渉, 清水 宏  日本皮膚科学会雑誌  120-  (3)  688  -688  2010/03  [Not refereed][Not invited]
  
• KRT17変異を持つ先天性爪甲厚硬症2型1家系における多発性脂腺嚢腫の遺伝学的・組織学的検討

  神田 真聡, 夏賀 健, 西江 渉, 秋山 真志, 長崎 暁理, 清水 忠道, 清水 宏  日本小児皮膚科学会雑誌  28-  (2)  198  -198  2009/11  [Not refereed][Not invited]
  
• 筋ジストロフィー合併型単純型表皮水疱症の1例

  大内 健嗣, 舩越 建, 谷川 瑛子, 石河 晃, 小堺 有史, 夏賀 健, 秋山 真志  日本小児皮膚科学会雑誌  28-  (2)  199  -199  2009/11  [Not refereed][Not invited]
  
• 先天性厚硬爪甲症と多発性脂腺嚢腫

  神田 真聡, 夏賀 健, 秋山 真志, 清水 宏  日本小児皮膚科学会雑誌  28-  (2)  211,3  -213,3  2009/11  [Not refereed][Not invited]
   

  先天性厚硬爪甲症(PC)は稀なケラチン病の一つであり、発症に関与するケラチン遺伝子により1型と2型(PC-2)に分類される。PCの臨床像と遺伝学的特徴を概説すると共に、PC-2に特徴的な所見である多発性脂腺嚢腫の形成機序について最新の知見を述べた。PC-2家系における脂腺嚢腫の遺伝学的および組織学的検討により、その形成には、遺伝子変異によって生じた異常なケラチン蛋白が正常に機能せずに凝集していることが関わっていることが示唆された。
• 【皮膚の病気のすべて】 皮膚の異常と病気 表皮水疱症

  新熊 悟, 夏賀 健, 清水 宏  からだの科学  (262)  51  -55  2009/08  [Not refereed][Not invited]
  
• 担癌患者に生じた自己免疫性水疱症の1例

  守内 玲寧, 阿部 理一郎, 夏賀 健, 乃村 俊史, 清水 宏, 三田村 卓, 櫻木 範明, 橋本 隆  日本皮膚科学会雑誌  119-  (2)  210  -210  2009/02  [Not refereed][Not invited]
  
• Speckled lentiginous nevus上に生じた悪性黒色腫の1例

  後藤 由香, 青柳 哲, 夏賀 健, 秦 洋郎, 猪熊 大輔, 澤村 大輔, 清水 宏, 月永 一郎, 深澤 雄一郎  日本皮膚科学会雑誌  119-  (1)  58  -58  2009/01  [Not refereed][Not invited]
  
• Perforating dermatosisの1例

  村瀬 修平, 阿部 理一郎, 野村 友希子, 夏賀 健, 立石 八寿貴, 冨田 幸希, 清水 宏, 松村 哲理  日本皮膚科学会雑誌  119-  (1)  60  -60  2009/01  [Not refereed][Not invited]
  
• Amicrobial pustulosis associated with autoimmune diseases

  夏賀 健, 澤村 大輔, 本間 英里奈, 乃村 俊史, 安部 将隆, 清水 宏, 村松 隆一  日本皮膚科学会雑誌  119-  (1)  62  -62  2009/01  [Not refereed][Not invited]
  
• 北海道がんセンターにおける過去13年間の悪性黒色腫87例の検討

  山根 尚子, 加藤 直子, 大澤 倫子, 柳 輝希, 皆川 英彦, 舟山 恵美, 夏賀 健  日本皮膚科学会雑誌  119-  (1)  66  -66  2009/01  [Not refereed][Not invited]
  
• 複数の皮膚悪性腫瘍に罹患し、ABCA12遺伝子の新規変異が同定された非水疱型先天性魚鱗癬様紅皮症の2例

  夏賀 健, 秋山 真志, 坂井 香織, 中桐 頼子, 西村 真智子, 安倍 将隆, 有田 賢, 阿部 由紀子, 小野塚 貴, 青柳 哲, 秦 洋郎, 小玉 和郎, 清水 宏, 加藤 直子, 氏家 英之, 冨田 幸希  日本皮膚科学会雑誌  119-  (1)  71  -71  2009/01  [Not refereed][Not invited]
  
• Anaplastic large cell lymphoma(ALCL)の1例

  山根 尚子, 加藤 直子, 柳 輝希, 大澤 倫子, 夏賀 健, 氏家 英之, 富田 幸希  日本皮膚科学会雑誌  119-  (1)  73  -73  2009/01  [Not refereed][Not invited]
  
• 皮膚に単発したInfantile myofibromatosisの1例

  菊地 一博, 阿部 理一郎, 新熊 悟, 濱坂 恵理香, 夏賀 健, 秦 洋郎, 立石 八寿貴, 柴田 雅彦, 冨田 幸希, 阿部 由紀子, 青柳 哲, 清水 宏, 向井 万起男  日本皮膚科学会雑誌  119-  (1)  75  -76  2009/01  [Not refereed][Not invited]
  
• 全身の膿疱

  小玉 和郎, 夏賀 健, 本間 英里奈, 乃村 俊史, 清水 宏, 澤村 大輔  日本皮膚科学会雑誌  118-  (10)  1985  -1985  2008/09  [Not refereed][Not invited]
  
• 北海道がんセンターにおける過去14年間の悪性黒色腫90例の検討

  山根 尚子, 加藤 直子, 大澤 倫子, 柳 輝希, 伊藤 幹, 西村 真智子, 皆川 英彦, 舟山 恵美, 林 利彦, 夏賀 健  日本皮膚科学会雑誌  118-  (10)  2024  -2024  2008/09  [Not refereed][Not invited]
  
• 非水疱型先天性魚鱗癬様紅皮症(NBCIE)患者に生じた悪性黒色腫と末梢T細胞性リンパ腫(PTCL)

  夏賀 健, 加藤 直子, 氏家 英之, 冨田 幸希  日本皮膚科学会雑誌  118-  (7)  1278  -1279  2008/06  [Not refereed][Not invited]
  
• 頭皮に生じた脂腺癌の1例

  冨田 幸希, 加藤 直子, 夏賀 健, 氏家 英之  皮膚科の臨床  50-  (4)  539  -541  2008/04  [Not refereed][Not invited]
   

  42歳男。左頭頂部に非対称性で2.8×1.5×1.5cmの淡紅色から黄色調の弾性硬で表面平滑な広茎性の腫瘤が発現し、徐々に増大して一部潰瘍を伴っていた。病理所見では真皮から皮下脂肪織に細胞塊が存在し、基底細胞様細胞と脂腺細胞への分化を示す細胞が混在し、胞巣中心部に塊状壊死を認めた。また免疫組織染色では抗ヒトMLH1抗体と抗ヒトMSH2抗体ともに陰性でMTS(Muir-Torre症候群)の可能性は低いと考えた。以上より脂腺癌で病期は皮膚有棘細胞癌の病期分類のT2 N0M0、Stage IIと診断した。治療は辺縁2cmマージンで下方は帽状腱膜上で腫瘍を切除し、分層植皮術を施行した。術後7ヵ月経過して再発転移は認めていない。
• 免疫グロブリン大量静注療法が著効した水疱性類天疱瘡の1例

  新熊 悟, 阿部 由紀子, 冨田 幸希, 夏賀 健, 氏家 英之, 阿部 理一郎, 秋山 真志, 清水 宏  日本皮膚科学会雑誌  118-  (5)  933  -937  2008/04  [Not refereed][Not invited]
   

  83歳女性。略全身の浮腫性紅斑と水疱、びらんを主訴に来院した。臨床、病理組織、免疫組織学的所見およびBP180ELISA値から水疱性類天疱瘡と診断した。ステロイドの全身投与(prednisolone,0.75mg/kg/日)を開始したが、内服開始後4週間の時点でも紅斑、水疱の新生を認めた。肺癌を併発しており全身状態が不良であることから、免疫抑制剤の併用や血漿交換療法はリスクが高いと考え、免疫グロブリン大量静注療法(400mg/kg/日、5日間)を施行した。投与直後より皮疹の著明な改善とBP180ELISA値の著明な低下を認め、その後ステロイドの減量を行うも皮疹の再燃は認めなかった。(著者抄録)
• Speckled lentiginous nevus上に生じた悪性黒色腫の1例

  後藤 由香, 青柳 哲, 夏賀 健, 秦 洋郎, 猪熊 大輔, 澤村 大輔, 月永 一郎, 深澤 雄一郎, 清水 宏  臨床皮膚科  62-  (1)  62  -65  2008/01  [Not refereed][Not invited]
   

  69歳,男性.出生時から腰部に帯状の巨大色素斑が存在し,3年前から色素斑上に黒色局面が出現した.病理組織学的には色素斑は単純黒子と境界部型の母斑細胞母斑が混在しており,また黒色局面はtumor thickness1.2mmの表在拡大型悪性黒色腫であった.センチネルリンパ節転移も認めたため,鼠径リンパ節郭清術と化学療法にて追加治療した.術後12ヵ月を経過して再発,転移を認めない.speckled lentiginous nevusは,稀ながら悪性黒色腫を生じる例もあり,慎重な経過観察が必要であると考えた.(著者抄録)
• Case report: A case of malignant melanoma on a speckled lentiginous nevus

  後藤 由香, 青柳 哲, 夏賀 健  臨床皮膚科  62-  (1)  62  -65  2008/01
• 皮膚科からみた口腔粘膜疾患 基礎と臨床

  夏賀 健, 清水 宏  老年歯科医学  22-  (2)  71  -75  2007/09  [Not refereed][Not invited]
  
• 免疫グロブリン大量静注療法が著効した水疱性類天疱瘡の1例

  新熊 悟, 阿部 由紀子, 冨田 幸希, 夏賀 健, 氏家 英之, 阿部 理一郎, 秋山 真志, 清水 宏  皮膚の科学  6-  (3)  319  -319  2007/06  [Not refereed][Not invited]
  
• 特異な角化性局面を契機にHIV感染症と診断した1例

  新熊 悟, 阿部 理一郎, 西村 真智子, 夏賀 健, 西江 渉, 浜坂 幸吉, 清水 宏  日本皮膚科学会雑誌  117-  (4)  710  -710  2007/03  [Not refereed][Not invited]
  
• 症例報告 HIV感染患者に生じた第2期梅毒の1例

  夏賀 健, 芝木 晃彦, 清水 宏  臨床皮膚科  60-  (9)  847  -849  2006/08  

  20歳男性.患者は同性愛者で1年前にHIV感染していた.今回,発熱から全身に皮疹が出現したため受診となった.所見では,躯幹・四肢・手掌・足底に径2mm大で鮮紅色,浮腫状で表面平滑な丘疹が散在性に認められた.検査所見ではCD4/CD8 0.46,RPP 64倍,TPLA 882.02TUで,腹部の丘疹の生検では,表皮は肥厚して軽度の海綿状態を呈し,真皮上層から表皮内にかけて炎症細胞が密に浸潤,部分的に液状変性も確認された.浸潤細胞はリンパ球,組織球からなり,多数の形質細胞を混じていた.抗treponema pallidum抗体により表皮内,真皮上層にらせん状の病原体を認め,HIV感染患者に生じた第2期梅毒と診断された.以上より,治療としてベンジルペニシリンベンザチンの8週間投与を行ったところ皮疹は消失し,抗体価はRPP 8倍,TPLA 191.0TUまで低下した
• Ultrastructural observations in a pigmented nevus in Hermansky-Pudlak syndrome (HPS) type 1: abnormal melanosome formation in nevus cells is of significant diagnostic value

  K Natsuga, M Akiyama, T Shimizu, T Suzuki, S Ito, Y Tomita, J Tanaka, H Shimizu  JOURNAL OF INVESTIGATIVE DERMATOLOGY  123-  (5)  A93  -A93  2004/11  [Not refereed][Not invited]
  

Presentations

• Revertant mosaicism  [Invited]

  Ken Natsuga

  EB Clinet Conference 2024  2024/10
• Revertant mosaicism therapy for EB  [Invited]

  Ken Natsuga

  International Epidermolysis Bullosa Symposium  2023/08
• Evaluation of revertant mosaicism in EB  [Invited]

  Ken Natsuga

  International EB Symposium  2023/05
• Physiology and pathology of the cutaneous basement membrane zone  [Invited]

  Ken Natsuga

  The 47th Annual Meeting of the Japanese Society for Investigative Dermatology  2022/12
• Biology of the cutaneous basement membrane zone: the state of the art  [Invited]

  Ken Natsuga

  The 48th Annual Meeting of the Taiwanese Dermatological Association (TDA)  2022/11
• Bursting the Bubble: Advances in the Diagnosis of Bullous Diseases  [Invited]

  Ken Natsuga

  4th Annual Meeting of Dermatopathology  2022/10
• Revertant mosaicism in EB  [Invited]

  Ken Natsuga

  International epidermolysis bullosa workshop  2022/08
• Diagnosis and treatment for genodermatosis in Japan  [Invited]

  Ken Natsuga

  2021 SATU Joint Research/SDGs Seminar  2021/08
• EB care and research in Japan  [Invited]

  Ken Natsuga

  International epidermolysis bullosa workshop  2021/01
• Epidermolysis bullosa: implications for stem cells and wound healing.  [Invited]

  Ken Natsuga

  JSID Asian Kisaragi-Juku 2020  2020/12
• Epidermal responses to physical stimuli  [Invited]

  Ken Natsuga

  Japan-Singapore International Skin Conference 2019  2019/04
• Organization of the Basement Membrane Zone: Implications for Epidermolysis Bullosa.  [Invited]

  Ken Natsuga

  115th Annual Meeting of the Japanese Dermatological Association (JDA)  2016/06
• Organization of the Basement Membrane Zone: Implications for Bullous Pemphigoid and Epidermolysis Bullosa.  [Invited]

  Ken Natsuga

  72nd annual meeting of the American Academy of Dermatology  2014/03
• Pathogenetic mechanisms in autoimmune skin fragility.  [Invited]

  Ken Natsuga

  40th annual SCUR meeting/ 6th joint meeting of SCUR and SSSR  2013/05

Association Memberships

• THE JAPANESE SOCIETY FOR INVESTIGATIVE DERMATOLOGY   THE JAPANESE DERMATOLOGICAL ASSOCIATION   日本人類遺伝学会   米国研究皮膚科学会   日本乾癬学会   日本白斑学会   皮膚の会   欧州研究皮膚科学会   日本臨床皮膚科医会   

Research Projects

• 表皮水疱症におけるリバータントスキンの診断法確立

  日本医療研究開発機構 難治性疾患実用化研究事業：

  Date (from‐to) : 2024/04 -2027/03 

  Author : 夏賀 健、高島翔太
• 表皮水疱症の創傷治癒遅延因子の同定とその克服

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  Date (from‐to) : 2023/04 -2026/03 

  Author : 夏賀 健
• 加齢に着目した皮膚の免疫自己寛容破綻機序の解明

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  Date (from‐to) : 2021/04 -2024/03 

  Author : 氏家 英之, 柳 輝希, 泉 健太郎, 夏賀 健, 岩田 浩明

   

  免疫系の老化、すなわち免疫老化によって自己免疫疾患のリスクが増加することが知られているが、詳細な機序の大部分は不明である。水疱性類天疱瘡（BP）は表皮基底膜部に存在するBP180やBP230に対する自己抗体によって生じる自己免疫性水疱症で、高齢者に好発する。我々は最近、一部の高齢マウスは表皮基底膜部に対する自己抗体を自然産生していることを発見した。本研究の目的は、BPをモデル疾患として“加齢に伴う免疫自己寛容の破綻機序を解明する”ことである。 まず、高齢マウスが産生する抗基底膜部自己抗体の標的抗原を同定するため、BP180、ラミニンγ1、ラミニンγ2、ラミニンβ3、ラミニンα3の各リコンビナントタンパクを作成した。高齢マウス（70週齢以上）の血清を採取し、作成したリコンビナントタンパクを基質としてウエスタンブロットを施行した。BP180リコンビナントタンパクを基質としたウエスタンブロット法で、約半数の高齢マウス血清中に抗BP180 IgM抗体が検出された。ラミニンγ1、ラミニンγ2、ラミニンβ3、ラミニンα3を基質としたウエスタンブロット法は陰性だった。 次に、加齢に伴いB細胞が抗基底膜部自己抗体を産生する機序を、制御性T細胞(Treg)とAge-associate B cells（ABCs）に着目して検討した。Treg欠損マウスでは野生型マウスに比較して、脾臓におけるABCsが増加していた。 また、薬剤投与により高齢マウスに病原性自己抗体を誘導できるかどうかを、近年BPの発症誘因として注目されているDPP-4阻害薬と抗PD-1抗体に着目して検討している。現在、高齢マウスにこれらの薬剤を投与中だが、今のところBP自己抗体の有意な誘導は見られていない。
• エピトープスプレディング現象に着目した水疱性類天疱瘡の病態解明

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  Date (from‐to) : 2020/04 -2023/03 

  Author : 西江 渉, 岩田 浩明, 泉 健太郎, 氏家 英之, 夏賀 健

   

  2020年度は、18名の水疱性類天疱瘡患者（DPP4阻害薬内服中の症例を含む）末梢血単核球中のB細胞をEBウイルスを用い形質転換した。樹立したLymphoblastoid cell lineの培養上清を用い、水疱性類天疱瘡抗原であるBP180タンパクへの自己抗体の反応性をELISA法で確認したところ、全ての患者さん由来細胞でIgGクラス自己抗体の反応性は確認できなかった。しかし、6名のLymphoblastoid cell line培養上清に坑BP180 IgMクラス自己抗体を確認したため、形質転換した細胞群から単一あるいはポリクローナルな状態での、坑BP180自己抗体産生細胞の採取をFACSソーティングで試みた。その結果、坑BP180自己抗体産生細胞数が極めて少なかったことと、蛍光標識BP180タンパクが細胞表面に結合した後のエンドサイトーシスに伴う蛍光の減弱のため、細胞の単離は上手くいかなかった。そこで、pH耐性緑色蛍光タンパクであるGamillusを融合したBP180タンパクを作製し、再度FACSソーティングを行うこととした。細胞を単離したのち、次世代シークエンサーによる自己抗体可変領域のアミノ酸配列の解読と、リコンビナント坑BP180自己抗体の作製、そしてマウスへ投与し病原性の確認を計画しそれぞれの予備実験を開始していたが、今後の研究を遂行できなくなったため、2021年以降の交付申請は辞退することにした。
• 表皮水疱症の治療最適化戦略

  日本医療研究開発機構：難治性疾患実用化研究事業

  Date (from‐to) : 2018/04 -2021/03 

  Author : 夏賀 健
• Novel bullous pemphigoid models induced by breaking tolerance to autoantigens.

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2016/04 -2019/03 

  Author : Nishie Wataru, IZUMI kentaro, TOYONAGA ellen

   

  The aim of this study is to develop bullous pemphigoid models for elucidating the pathomechanisms and producing novel therapies. We have established several novel bullous pemphigoid models by immunizing recombinant collagen XVII (COL17) proteins. Notably, monoclonal antibodies derived from the immunized mice disclosed that epitopes and subclasses of the monoclonal antibodies were associated with phenotypes of skin disease.
• Establishment of cicatricial alopecia animal models

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2016/04 -2019/03 

  Author : Nakamura Hideki

   

  Cicatricial alopecia is a type of hair loss characterized by loss of hair follicles and surrounding fibrosis. The aim of this study was to elucidate the mechanism of cicatricial alopecia by analyzing the hair follicles in the blistered skin of mice in detail. A major finding was a delay in hair follicle development/cycling at the re-epithelialized wounds after blistering. At the same time, infiltration of inflammatory cells was poor at the site where several layers of epithelization were completed, and there was no difference compared to the intact site. Taken together, these results indicate that the early stage of cicatricial alopecia involves less inflammation, with the proliferation of epithelial cells required for hair follicle growth directed toward re-epithelialization.
• Dynamics of epithelial stem cells upon wounding

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)

  Date (from‐to) : 2017/04 -2019/03 

  Author : Natsuga Ken, Watanabe Mika, Fujimura Yu

   

  In wounds in which the epidermis was selectively removed, there was 1 -3 reepithelialization at 24 hours after wounding as visible through HE staining. The basal layer of the epidermis expressing α6 integrin was present at this point, and cells in the re-epithelialized layer were stained with pan-cytokeratin. Loricrin staining, showing differentiation into epidermal granular layer, was not seen 48 hours after wound creation but was confirmed 72 hours later. HE staining also shows no granular layer at 48 hours after wounding, but at 72 hours after wounding, the final reepithelialization is complete. Lineage tracing also revealed that daughter cells of hair follicle upper stem cells are involved in wound reepithelialization.
• Pathogenesis of bullous pemphigoid: a study using class-switched monoclonal antibodies.

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2016/04 -2018/03 

  Author : Nishie Wataru

   

  Bullous pemphigoid (BP) is an autoimmune blistering skin disease, in which autoantibodies target collagen XVII (COL17) in the skin. Here we produced recombinant monoclonal antibodies (mAb) directing COL17 with different isotypes including IgG1, IgG2a, IgA and IgE. These mAbs have identical variable regions. Passive transfer of these mAbs into mice induced activation of complements when IgG2a but not IgG1 mAbs were injected. However, there was no clinical differences were observed in complement-activating and complement-non-activating recipient mice. These results suggest that activation of complements is not enough to induce blistering skin disease in mice.
• Treatment of epidermolysis bullosa with revertant keratinocyte-derived iPS cells

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2015/04 -2018/03 

  Author : Fujita Yasuyuki, MATSUMURA WAKANA, NAKAYAMA CHIHIRO

   

  Cell-based therapies such as allogeneic mesenchymal stem/stromal cells (MSCs) transplantation for recessive dystrophic epidermolysis bullosa (RDEB) were explored. However, some hurdles exist to come over in current MSC-based therapies; limited proliferation and limited cell survival. To solve these problems, we focused on keratinocyte-derived induced pluripotent stem cells (iPSCs). Keratinocytes from intact areas of patients might have revertant mosaicism where the pathogenic mutations are spontaneously corrected. In this study, we succeeded to develop MSCs from keratinocytes-derived iPSCs (KC-iPSC-MSCs) of normal human and a RDEB patient. Keratinocytes-derived iPSCs (KC-iPSCs) were cultured with activin A, BIO and BMP4 to induct mesoderm lineage, and then cultured with bFGF and EGF. In 14 days, spindle-shaped cells appeared which showed mesenchymal differentiation. Furthermore, the intravenous injection of the KC-iPSC-MSCs into wounded mice induced human type VII collagen.
• Prevention of autoantigen depletion for bullous pemphigoid treatment

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)

  Date (from‐to) : 2015/04 -2017/03 

  Author : Natsuga Ken, Watanabe Mika, Fujimura Yu

   

  Immune system works against external pathogens such as viruses and bacteria. Once immune system does not function well, it targets our body cells. This phenomenon is called "autoimmunity". Bullous pemphigoid, an autoimmune disease in which skin attachment molecules are targeted, shows skin blisters on the whole body. We analyzed how skin cells respond to the treatment with antibodies against skin. As a result, the expression levels of molecules associated with intracellular trafficking were increased.
• Induction of tolerance to bullous pemphigoid autoantigen by oral administration of antigenic peptides.

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2015/04 -2016/03 

  Author : Nishie Wataru, NATSUGA KEN

   

  Bullous pemphigoid (BP) is the major autoimmune blistering disease in which autoantibodies mainly target NC16A domain of collagen XVII (COL17). BP may be fatal because majority of patients are treated with systemic immmune suppressive drugs; thus, disease-specific therapies are required. In this study, we aimed at the fact that BP autoantibodies mainly target NC16A domain, and hypothesized that oral treatment with NC16A peptides may induce tolerance to COL17. We showed that oral treatment of NC16A peptides can suppress development of autoantibodies directing COL17 in mice, although statistical significance was not achieved. However, this finding must be very helpful to establish novel disaese-specific therapies for BP in the near future.
• Elucidating roles of plasmin in blister formation of bullous pemphigoid

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2013/04 -2016/03 

  Author : Nakamura Hideki, NISHIE Wataru, NATSUGA Ken, SHIMIZU Hiroshi

   

  Bullous pemphigoid (BP) is an autoimmune blistering disease. The aim of this study is to address the pathogenic roles of plasmin, a protease present in blister fluids of BP, which is known to cleave COL17. We first identified cleavage sites of COL17 by plasmin, and based on this data, cleavage site-specific antibodies were generated. The antibodies revealed that COL17 is actually cleaved by plasmin in lesional skin of certain numbers of BP patients.
• Production of a novel blistering disease model by regulating expression of collagen XVII

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2012/04 -2016/03 

  Author : Nishie Wataru, UJIIE HIDEYUKI, SHINKUMA SATORU, NATSUGA KEN

   

  Collagen XVII (COL17) is a hemidesmosomal molecule which keeps adhesion between the epidermis and dermis. Loss of CO17 expression due to mutation in the COL17A1 leads to blistering disease, epidermolysis bullosa (EB); in addition, autoimmunity to COL17 results in autoimmune blistering disorder bullous pemphigoid (BP). The purpose of this study is to produce a novel blistering model by regulating COL17 expression based on Tet-on system. Transgenic mice carrying human COL17 cDNA driven under the CMV promoter with Tet response element and Tet transactivator sequences driven under the K14 promoter were produced. Upon oral administration of doxycycline (DOX), the Tg mice started to express human COL17 in basal keratinocytes, following by production of IgG autoantibodies directing human COL17. The Tg mice must be useful tool for elucidating pathogenesis of EB and BP, as well as for establishing therapies for these orphan diseases.
• Generation of mosaic model of epidermolysis bullosa by inactivation of X chromosome.

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2014/04 -2015/03 

  Author : NISHIE Wataru, NATSUGA Ken

   

  The popups of this study is to generate a mosaic model of epidermolysis bullosa by inactivation of X chromosome. We have generated transgenic (Tg) mice with tetracycline response plasmid expressing human type XVII collagen (COL17) cDNA, and then crossed with the Tg mice expressing reverse tetracycline transactivator driven under keratin 14 promoter on X-chromosome. When primary keratinocytes from the double Tg mice were treated with doxycycline in culture, increased human COL17 gene expression was observed; however, COL17 protein expression was not induced. These results indicate that it may be challenging for this method to obtain mice with high inducible level of COL17, which is needed to be improved.
• Modulation of skin microbiota through basememt membrane

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2013/04 -2014/03 

  Author : SHIMIZU Hiroshi, NISHIE Wataru, NATSUGA Ken

   

  Skin microbiota of mice deficient for skin basement membrane protein (BMZ-deficient mice) were analyzed. At P1, BMZ-deficient mice had less abundant skin microbiota compared with the control group mice while the quantity of skin bacteria was comparable between BMZ-deficient mice and the control mice at P4. Visualization of skin microbiota using FISH methods revealed that bacterial signals were located mainly in the cornified layers of BMZ-deficient mice and the control mice. The specificity of the probe to detect bacterial signals was confirmed by the negative FISH signals using a probe reacting with fungal species which are not found in skin. Further analyses will be done on microbiota of adult BMZ-deficient mice skin and human epidermolysis bullosa patients' skin.