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A homozygous single T deletion found in the GGCX gene with PXE-like phenotypes
Abstracts / Journal of Dermatological Science 84 (2016) e1–e88
P07-08
P07-09
The role of the hairless (hr) gene in the development of atopic dermatitis-like pruritus caused by feeding a special diet to mice
HLA-C*12:02 is a susceptibility factor in late-onset type of psoriasis in Japanese
Masanori Fujii 1,∗ , Fumiko Endo-Okuno 1 , Asuka Iwai 1 , Keisuke Doi 1 , Yoshiki Matsumoto 1 , Rieko Matsui 1 , Yumeka Yamada 1 , Naoki Inagaki 2 , Takeshi Nabe 1,3 , Susumu Ohya 1 1
Department of Pharmacology, Division of Pathological Sciences, Kyoto Pharmaceutical University, Japan 2 Laboratory of Pharmacology, Department of Bioactive Molecules, Gifu Pharmaceutical University, Japan 3 Laboratory of Toxicology, Faculty of Pharmaceutical Sciences, Setsunan University, Japan Atopic dermatitis (AD) is a highly pruritic inflammatory skin disease that can be regulated by various genetic factors. We have shown that HR-1 hairless mice fed a commercial special diet (named HR-AD) develop AD-like pruritic skin inflammation that is caused by deficiency of polyunsaturated fatty acids (PUFAs) (Fujii et al., Exp Dermatol, 2013). In the present study, we examined the influence of genetic background on the diet induced AD-like pruritus. Five different strains of mice (HR-1, BALB/c, C57BL/6, ddY and ICR) were given a normal diet or the special diet for 12 weeks and AD-related symptoms were assessed. Among the five strains of mice fed the special diet, HR-1 mice were most severely affected in terms of decreased skin hydration, reduced skin barrier function, itch-related scratching and an increase in dermal mast cells. Interestingly, the mast cell hyperplasia was observed even in normal diet-fed HR-1 mice. Because only HR-1 mice carry a mutation in the hairless (hr) gene, we hypothesized that the hr mutation is responsible for the high susceptibility to develop AD-related symptoms in HR-1 mice. To prove this, we newly generated a hairless congenic strain that carries the same hr mutation on ICR background (called hr-ICR). HR-1 mice and hr-ICR mice showed similarly dry skin symptoms and itch-related scratching induced by the special diet feeding, indicating that the hr mutation promotes the development of these symptoms. On the other hand, the mast cell hyperplasia was not fully reproduced in hr-ICR mice, suggesting that genetic factor(s) other than the hr mutation is also involved in it. Taken together, our study demonstrates that mutation of the hairless (hr) gene drives AD-like pruritus caused by PUFA deficiency in mice. http://dx.doi.org/10.1016/j.jdermsci.2016.08.153
e49
Tomotaka Mabuchi 1,∗ , Tami Ota 1 , Yasuaki Manabe 1 , Norihiro Ikoma 1 , Akira Ozawa 1 , Tadashi Terui 2 , Shigaku Ikeda 3 , Hidetoshi Inoko 4 , Akira Oka 5 1 Department of Dermatology, Tokai University School of Medicine, Kanagawa, Japan 2 Department of Dermatology, Nihon University School of Medicine, Tokyo, Japan 3 Department of Dermatology, Juntendo University School of Medicine, Tokyo, Japan 4 Molecular Life Science, Tokai University School of Medicine, Kanagawa, Japan 5 The Institute of Medical Science, Tokai University School of Medicine, Kanagawa, Japan
Background: HLA-C locus on chromosome 6p21.33 remains the strongest susceptibility candidate locus in psoriasis. The strong association between psoriasis and HLA-Cw6 allele has been well known in various races. It is known that psoriatic patients with early onset are more likely to be familial and associated with HLACw6, but it was not confirmed in Japanese yet. On the other hand, no specific late-onset type of psoriasis susceptibility loci, including HLA-C alleles has been reported. Methods: Four hundred forty six unrelated Japanese patients with psoriasis vulgaris and 557 sex- and age-matched unrelated Japanese healthy controls were investigated by genotyping. Results: The strong association between HLA-C*06:02 allele and early-onset type (OR = 13.9, 95% CI: 5.72–40.7, p = 3.91E−13) was observed, but late-onset type could not significantly demonstrated association with HLA-C*06:02 allele (OR = 2.38, 95% CI: 0.57–9.45, p = 0.167). By contrast, late-onset type of psoriasis indicated significant association with HLA-C*12:02 allele (OR = 1.91, 95% CI: 1.38–2.63, p = 5.48E–05), although there was no significant association with early-onset type of psoriasis. Conclusion: We confirmed the association between early-onset type of psoriasis with HLA-C*06:02 allele in Japanese, however, it was weaker than other populations. In addition, we detected the association between late-onset type of psoriasis with HLA-C*12:02 allele in Japanese. http://dx.doi.org/10.1016/j.jdermsci.2016.08.154 P07-10 A homozygous single T deletion found in the GGCX gene with PXE-like phenotypes Kosuke Yoshimi ∗ , Yumi Okubo, Susumu Ikehara, Yuta Koike, Yutaka Kuwatsuka, Atsushi Utani The Department of Dermatology, Nagasaki University, Nagasaki, Japan Pseudoxanthoma elasticum like disorder with multiple coagulation deficiency (tentatively named GGCX syndrome) is known to be caused by GGCX (gamma-glutamyl carboxylase) gene mutation. GGCX is an enzyme that carboxylates glutamate residue in a group of proteins including anti-calcification molecules (matrix gla protein (MGP), alpha-fetuin, etc.), and vitamine K dependent coagulation factors. We report a 55 years old Japanese male with GGCX syndrome shows pseudoxanthoma (PXE)-like phenotype such as cutis laxa, calcification of elastic fibers in the dermis, peripapillary atrophy of the retina, and coagulation deficiency. Furthermore
e50
Abstracts / Journal of Dermatological Science 84 (2016) e1–e88
he has been suffered from dysarthria and motor ataxia suspect of spinocerebellar degeneration as unreported phenotype. We found homozygous deletion of T within C-terminal end. It produces frameshift and yields 77 amino acid larger molecule of GGCX. Abnormal GGCX molecule located in rER in patient fibroblasts, which was similar pattern in normal fibroblasts. However one of GGCX substrate MGP was under carboxylated demonstrated by western blotting using carboxylation dependent antibody to MGP. These data suggest that this mutation induced loss of function of GGCX resulted in elastic fiber calcification as well as coagulation deficiency. The reason why he had symptoms due to possible spinocerebellar degeneration remains to be elucidated. http://dx.doi.org/10.1016/j.jdermsci.2016.08.155 P07-11 Postzygotic KRAS G12C mutation, associated with various malignancies, is also underling epidermal nevus syndrome Satomi Igawa 1,∗ , Masaru Honma 1 , Masako Minami-Hori 1 , Etsushi Tsuchida 2 , Hajime Iizuka 1 , Akemi Ishida-Yamamoto 1
including wrinkling, by activating the metalloproteinases (MMP1) that break down type I collagen (COL1A1). Mushrooms gathered from the wild flora have always been included in the diet of humans for centuries due to their specific taste as well as their medicinal and curative properties. P. cornucopiae, the golden oyster mushroom, is an edible mushroom and commonly found in East Asia. The anticancer and hepatoprotective effect of P. cornucopiae has been reported, but it has rarely been applied in skin care. This study was performed to evaluate the antiaging effect of P. cornucopiae extract (PCE). The antioxidant activities of PCE extract were determined by using five different methods, namely, DPPH and ABTS radical scavenging assay, ferric reducing antioxidant power (FRAP) assay and ORAC assay. Also antioxidant effect of PCE was evaluated by estimating its protective effects against the oxidative stress induced by H2 O2 on human dermal fibroblasts. PCE showed radical scavenging and cytoprotective activity against oxidative stress in a dose dependent manner. PCE additionally prevented the elevation of MMP-1 and reduction of COL1A1 induced by H2 O2 . Theses finding indicated that PCE protects human skin fibroblasts from H2 O2 -induced damage by antioxidant activity and therefore can be used as an antiaging agent. http://dx.doi.org/10.1016/j.jdermsci.2016.08.157
1
Department of Dermatology, Asahikawa Medical University, Asahikawa, Japan 2 Department of Pediatrics, Asahikawa Medical University, Asahikawa, Japan A KRAS c.34G>T p.G12C point mutation has been reported in various carcinomas and sarcomas, but it has never been reported in epidermal nevus syndrome (ENS), a heterogeneous group of congenital disorders characterized by the presence of epidermal nevi associated with systemic involvement. Keratinocytic epidermal nevus syndrome and nevus sebaceous/Schimmelpenning syndrome are included in this group. Recently, keratinocytic epidermal nevus (KEN) and sebaceous nevus (SN) were found to be caused by postzygotic mutations in several genes related to cell proliferation, among which are HRAS and KRAS. Interestingly, the same point mutation was reported in KEN and SN. We report a case of a three-year-old Japanese girl with two types of nevi on the left side of her body following Blaschko lines present since birth: yellowish plaques suggesting SN on the head and face, and dark brownish verrucous plaques suggesting KEN on the trunk and lower extremity. Detailed examination revealed delayed eruption of the left teeth and a left cerebral arachnoid cyst. Mutation analysis showed a KRAS c.34G>T p.G12C point mutation in genomic DNA derived from her lesional epidermis but not in her nonlesional epidermis or peripheral blood leukocytes. As far as we are aware, this is the first case of ENS caused by postzygotic KRAS G12C mutation. No malignant tumors have developed in the patient up to the present age of three years, but regular follow-up is warranted. http://dx.doi.org/10.1016/j.jdermsci.2016.08.156 P07-12 Antiaging effect of pleurotus cornucopiae extracts Kum Hyun Woo ∗ , Seoung Woo Shin, Eunsun Jung, Deok Hoon Park, Min kyung Kim, Rin a Lee Life Science Institute, Seongnam City, Gyeonggi Do, Republic of Korea Oxidative stress is commonly involved in the pathogenesis of skin damage induced by these factors, causing skin aging,
Category 8(P08): Tissue Regeneration/Stem Cell and Wound Healing P08-01[I-1] A cell polarity protein, aPKC, is essential for maintaining hair follicle stem cell quiescence and hair follicle regeneration Shin-Ichi Osada 1,∗ , Naoko Minematsu 1 , Fumino Oda 1 , Kazunori Akimoto 2 , Seiji Kawana 1 , Shigeo Ohno 2 1
Department of Dermatology, Nippon Medical School, Tokyo, Japan 2 Department of Molecular Biology, Yokohama City University Graduate School of Medical Science, Yokohama, Kanagawa, Japan The atypical protein kinase C (aPKC)-PAR complex regulates the formation of tight junctions and apico-basal epithelial polarity. To examine the role of this complex in the epidermis, we generated mutant mice harboring epidermal specific deletion of aPKC (cKO), a major component of the aPKC-PAR complex. The mutant mice exhibited abnormal hair cycling, progressive losses of pelage hairs and vibrissae, and altered differentiation into the epidermis and sebaceous gland. We found that in the aPKC cKO mice hair follicle stem cell (HFSC) quiescence was lost, as revealed by the decreased expression level of quiescence-inducing factors (Fgf18 and Bmp6), produced in Keratin 6-positive bulge stem cells. The loss of quiescence dysregulated the bulge stem cell marker expression and led to the increase in Lrig1-positive cells, inducing hyperplasia of the interfollicular epidermis and sebaceous glands, and drove an increase in Lef1-positive matrix cells, causing a prolonged anagenlike phase. Persistent bulge stem cell activation led to a gradual depletion of CD34- and integrin ␣6-positive HFSC reservoirs. Moreover, hair follicle regeneration after wounding was disturbed in the mutant epidermis, where the expression of Wnt ligands and their downstream targets were severely suppressed. These results suggest aPKC regulates signaling pathways implicated in HFSC quiescence and hair follicle regeneration. http://dx.doi.org/10.1016/j.jdermsci.2016.08.158
P07-08
P07-09
The role of the hairless (hr) gene in the development of atopic dermatitis-like pruritus caused by feeding a special diet to mice
HLA-C*12:02 is a susceptibility factor in late-onset type of psoriasis in Japanese
Masanori Fujii 1,∗ , Fumiko Endo-Okuno 1 , Asuka Iwai 1 , Keisuke Doi 1 , Yoshiki Matsumoto 1 , Rieko Matsui 1 , Yumeka Yamada 1 , Naoki Inagaki 2 , Takeshi Nabe 1,3 , Susumu Ohya 1 1
Department of Pharmacology, Division of Pathological Sciences, Kyoto Pharmaceutical University, Japan 2 Laboratory of Pharmacology, Department of Bioactive Molecules, Gifu Pharmaceutical University, Japan 3 Laboratory of Toxicology, Faculty of Pharmaceutical Sciences, Setsunan University, Japan Atopic dermatitis (AD) is a highly pruritic inflammatory skin disease that can be regulated by various genetic factors. We have shown that HR-1 hairless mice fed a commercial special diet (named HR-AD) develop AD-like pruritic skin inflammation that is caused by deficiency of polyunsaturated fatty acids (PUFAs) (Fujii et al., Exp Dermatol, 2013). In the present study, we examined the influence of genetic background on the diet induced AD-like pruritus. Five different strains of mice (HR-1, BALB/c, C57BL/6, ddY and ICR) were given a normal diet or the special diet for 12 weeks and AD-related symptoms were assessed. Among the five strains of mice fed the special diet, HR-1 mice were most severely affected in terms of decreased skin hydration, reduced skin barrier function, itch-related scratching and an increase in dermal mast cells. Interestingly, the mast cell hyperplasia was observed even in normal diet-fed HR-1 mice. Because only HR-1 mice carry a mutation in the hairless (hr) gene, we hypothesized that the hr mutation is responsible for the high susceptibility to develop AD-related symptoms in HR-1 mice. To prove this, we newly generated a hairless congenic strain that carries the same hr mutation on ICR background (called hr-ICR). HR-1 mice and hr-ICR mice showed similarly dry skin symptoms and itch-related scratching induced by the special diet feeding, indicating that the hr mutation promotes the development of these symptoms. On the other hand, the mast cell hyperplasia was not fully reproduced in hr-ICR mice, suggesting that genetic factor(s) other than the hr mutation is also involved in it. Taken together, our study demonstrates that mutation of the hairless (hr) gene drives AD-like pruritus caused by PUFA deficiency in mice. http://dx.doi.org/10.1016/j.jdermsci.2016.08.153
e49
Tomotaka Mabuchi 1,∗ , Tami Ota 1 , Yasuaki Manabe 1 , Norihiro Ikoma 1 , Akira Ozawa 1 , Tadashi Terui 2 , Shigaku Ikeda 3 , Hidetoshi Inoko 4 , Akira Oka 5 1 Department of Dermatology, Tokai University School of Medicine, Kanagawa, Japan 2 Department of Dermatology, Nihon University School of Medicine, Tokyo, Japan 3 Department of Dermatology, Juntendo University School of Medicine, Tokyo, Japan 4 Molecular Life Science, Tokai University School of Medicine, Kanagawa, Japan 5 The Institute of Medical Science, Tokai University School of Medicine, Kanagawa, Japan
Background: HLA-C locus on chromosome 6p21.33 remains the strongest susceptibility candidate locus in psoriasis. The strong association between psoriasis and HLA-Cw6 allele has been well known in various races. It is known that psoriatic patients with early onset are more likely to be familial and associated with HLACw6, but it was not confirmed in Japanese yet. On the other hand, no specific late-onset type of psoriasis susceptibility loci, including HLA-C alleles has been reported. Methods: Four hundred forty six unrelated Japanese patients with psoriasis vulgaris and 557 sex- and age-matched unrelated Japanese healthy controls were investigated by genotyping. Results: The strong association between HLA-C*06:02 allele and early-onset type (OR = 13.9, 95% CI: 5.72–40.7, p = 3.91E−13) was observed, but late-onset type could not significantly demonstrated association with HLA-C*06:02 allele (OR = 2.38, 95% CI: 0.57–9.45, p = 0.167). By contrast, late-onset type of psoriasis indicated significant association with HLA-C*12:02 allele (OR = 1.91, 95% CI: 1.38–2.63, p = 5.48E–05), although there was no significant association with early-onset type of psoriasis. Conclusion: We confirmed the association between early-onset type of psoriasis with HLA-C*06:02 allele in Japanese, however, it was weaker than other populations. In addition, we detected the association between late-onset type of psoriasis with HLA-C*12:02 allele in Japanese. http://dx.doi.org/10.1016/j.jdermsci.2016.08.154 P07-10 A homozygous single T deletion found in the GGCX gene with PXE-like phenotypes Kosuke Yoshimi ∗ , Yumi Okubo, Susumu Ikehara, Yuta Koike, Yutaka Kuwatsuka, Atsushi Utani The Department of Dermatology, Nagasaki University, Nagasaki, Japan Pseudoxanthoma elasticum like disorder with multiple coagulation deficiency (tentatively named GGCX syndrome) is known to be caused by GGCX (gamma-glutamyl carboxylase) gene mutation. GGCX is an enzyme that carboxylates glutamate residue in a group of proteins including anti-calcification molecules (matrix gla protein (MGP), alpha-fetuin, etc.), and vitamine K dependent coagulation factors. We report a 55 years old Japanese male with GGCX syndrome shows pseudoxanthoma (PXE)-like phenotype such as cutis laxa, calcification of elastic fibers in the dermis, peripapillary atrophy of the retina, and coagulation deficiency. Furthermore
e50
Abstracts / Journal of Dermatological Science 84 (2016) e1–e88
he has been suffered from dysarthria and motor ataxia suspect of spinocerebellar degeneration as unreported phenotype. We found homozygous deletion of T within C-terminal end. It produces frameshift and yields 77 amino acid larger molecule of GGCX. Abnormal GGCX molecule located in rER in patient fibroblasts, which was similar pattern in normal fibroblasts. However one of GGCX substrate MGP was under carboxylated demonstrated by western blotting using carboxylation dependent antibody to MGP. These data suggest that this mutation induced loss of function of GGCX resulted in elastic fiber calcification as well as coagulation deficiency. The reason why he had symptoms due to possible spinocerebellar degeneration remains to be elucidated. http://dx.doi.org/10.1016/j.jdermsci.2016.08.155 P07-11 Postzygotic KRAS G12C mutation, associated with various malignancies, is also underling epidermal nevus syndrome Satomi Igawa 1,∗ , Masaru Honma 1 , Masako Minami-Hori 1 , Etsushi Tsuchida 2 , Hajime Iizuka 1 , Akemi Ishida-Yamamoto 1
including wrinkling, by activating the metalloproteinases (MMP1) that break down type I collagen (COL1A1). Mushrooms gathered from the wild flora have always been included in the diet of humans for centuries due to their specific taste as well as their medicinal and curative properties. P. cornucopiae, the golden oyster mushroom, is an edible mushroom and commonly found in East Asia. The anticancer and hepatoprotective effect of P. cornucopiae has been reported, but it has rarely been applied in skin care. This study was performed to evaluate the antiaging effect of P. cornucopiae extract (PCE). The antioxidant activities of PCE extract were determined by using five different methods, namely, DPPH and ABTS radical scavenging assay, ferric reducing antioxidant power (FRAP) assay and ORAC assay. Also antioxidant effect of PCE was evaluated by estimating its protective effects against the oxidative stress induced by H2 O2 on human dermal fibroblasts. PCE showed radical scavenging and cytoprotective activity against oxidative stress in a dose dependent manner. PCE additionally prevented the elevation of MMP-1 and reduction of COL1A1 induced by H2 O2 . Theses finding indicated that PCE protects human skin fibroblasts from H2 O2 -induced damage by antioxidant activity and therefore can be used as an antiaging agent. http://dx.doi.org/10.1016/j.jdermsci.2016.08.157
1
Department of Dermatology, Asahikawa Medical University, Asahikawa, Japan 2 Department of Pediatrics, Asahikawa Medical University, Asahikawa, Japan A KRAS c.34G>T p.G12C point mutation has been reported in various carcinomas and sarcomas, but it has never been reported in epidermal nevus syndrome (ENS), a heterogeneous group of congenital disorders characterized by the presence of epidermal nevi associated with systemic involvement. Keratinocytic epidermal nevus syndrome and nevus sebaceous/Schimmelpenning syndrome are included in this group. Recently, keratinocytic epidermal nevus (KEN) and sebaceous nevus (SN) were found to be caused by postzygotic mutations in several genes related to cell proliferation, among which are HRAS and KRAS. Interestingly, the same point mutation was reported in KEN and SN. We report a case of a three-year-old Japanese girl with two types of nevi on the left side of her body following Blaschko lines present since birth: yellowish plaques suggesting SN on the head and face, and dark brownish verrucous plaques suggesting KEN on the trunk and lower extremity. Detailed examination revealed delayed eruption of the left teeth and a left cerebral arachnoid cyst. Mutation analysis showed a KRAS c.34G>T p.G12C point mutation in genomic DNA derived from her lesional epidermis but not in her nonlesional epidermis or peripheral blood leukocytes. As far as we are aware, this is the first case of ENS caused by postzygotic KRAS G12C mutation. No malignant tumors have developed in the patient up to the present age of three years, but regular follow-up is warranted. http://dx.doi.org/10.1016/j.jdermsci.2016.08.156 P07-12 Antiaging effect of pleurotus cornucopiae extracts Kum Hyun Woo ∗ , Seoung Woo Shin, Eunsun Jung, Deok Hoon Park, Min kyung Kim, Rin a Lee Life Science Institute, Seongnam City, Gyeonggi Do, Republic of Korea Oxidative stress is commonly involved in the pathogenesis of skin damage induced by these factors, causing skin aging,
Category 8(P08): Tissue Regeneration/Stem Cell and Wound Healing P08-01[I-1] A cell polarity protein, aPKC, is essential for maintaining hair follicle stem cell quiescence and hair follicle regeneration Shin-Ichi Osada 1,∗ , Naoko Minematsu 1 , Fumino Oda 1 , Kazunori Akimoto 2 , Seiji Kawana 1 , Shigeo Ohno 2 1
Department of Dermatology, Nippon Medical School, Tokyo, Japan 2 Department of Molecular Biology, Yokohama City University Graduate School of Medical Science, Yokohama, Kanagawa, Japan The atypical protein kinase C (aPKC)-PAR complex regulates the formation of tight junctions and apico-basal epithelial polarity. To examine the role of this complex in the epidermis, we generated mutant mice harboring epidermal specific deletion of aPKC (cKO), a major component of the aPKC-PAR complex. The mutant mice exhibited abnormal hair cycling, progressive losses of pelage hairs and vibrissae, and altered differentiation into the epidermis and sebaceous gland. We found that in the aPKC cKO mice hair follicle stem cell (HFSC) quiescence was lost, as revealed by the decreased expression level of quiescence-inducing factors (Fgf18 and Bmp6), produced in Keratin 6-positive bulge stem cells. The loss of quiescence dysregulated the bulge stem cell marker expression and led to the increase in Lrig1-positive cells, inducing hyperplasia of the interfollicular epidermis and sebaceous glands, and drove an increase in Lef1-positive matrix cells, causing a prolonged anagenlike phase. Persistent bulge stem cell activation led to a gradual depletion of CD34- and integrin ␣6-positive HFSC reservoirs. Moreover, hair follicle regeneration after wounding was disturbed in the mutant epidermis, where the expression of Wnt ligands and their downstream targets were severely suppressed. These results suggest aPKC regulates signaling pathways implicated in HFSC quiescence and hair follicle regeneration. http://dx.doi.org/10.1016/j.jdermsci.2016.08.158