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TRPS1 haploinsufficiency results in increased STAT3 and SOX9 mRNA expression in hair follicles in trichorhinophalangeal syndrome
Abstracts / Journal of Dermatological Science 84 (2016) e1–e88
is also indispensable for maintenance of skin epidermis. To test the hypothesis, HPEK were irradiated with UVB. UVB irradiation stimulated BNIP3 expression and cleavage of caspase3. Surprisingly, suppression of BNIP3 expression induced by UVB irradiation caused a further increase of the cleaved caspase3 protein level, suggesting that BNIP3 has a protective effect against UVB-induced apoptosis in keratinocytes. Overall, our data shed light on functions of BNIP3, an inducer of autophagy, in both differentiation and maintenance of epidermal keratinocytes. http://dx.doi.org/10.1016/j.jdermsci.2016.08.175 P09-07 Plasma cytoid dendritic cells as a key player in the induction of alopecia areata Taisuke Ito ∗ , Takahiro Suzuki, Jun-ichi Sakabe, Atsuko Funakoshi, Yoshiki Tokura Department of Dermatology, Hamamatsu University School of Medicine, Japan Recently, the pathomechanism of alopecia areata (AA) has been thought to be a tissue-specific autoimmune disease. So far, interferon (IFN)-␥ has been regarded as the most important key cytokine that may induce the collapse of HF immune privilege, apoptosis and the upregulation of CXCL10 in C3H/HeJ AA mouse. In this study, we focused on the role of type I IFN, IFN-␣, in the initiation of AA. We previously reported seven cases of AA, including both new onset and recurrent AA, following swine flu viral infection. After viral infection, Th1-mediated immune responses are generally induced by an overproduction of interferons (IFNs). We also showed AA patients just after PEG-IFN-␣2b therapy. In this study, immunohistochemical staining revealed that IFN-␣ producing plasmacytoid dendritic cells (pDCs) densely distributed around HFs in not only lesion skin but also non-lesional skin obtained from C3H/HeJ mice with AA. Flowcytometric analysis showed the increased frequency of pDCs in skin-infiltrated cells of non-lesional skin from C3H/HeJ mice with AA. Real-time PCR also showed higher expression of IFN␣2 and IFN-␣4 mRNA in C3H/HeJ mouse with AA. In vitro study, IFN-␣ inhibited the hair elongation of cultured murine vibrissae in Willium’s E medium by Philpott’s model. In addition, H-2k and CXCL10 expression were upregulated in cultured murine vibrissae by IFN-␣. Imiquimod (IMQ) is the potent inducer of IFN-␣ via TLR7/9 on pDCs. IMQ applied C3H/HeJ mouse showed retardation of hair cycle stage compared to control mouse with infiltration of SiglecH+ pDCs with IFN-␣ expression by immunohistochemical staining. In conclusion, pDCs may play an important role for the initiation of AA by over expression of IFN-␣ via pDCs. http://dx.doi.org/10.1016/j.jdermsci.2016.08.176 P09-08 Human scalp-derived fibroblasts modulate FGF expression profile to be putatively folliculogenic in response to WNT activation Misaki Ise ∗ , Aki Tsukashima, Masayuki Amagai, Manabu Ohyama Department of Dermatology, Keio University School of Medicine, Tokyo, Japan Hair follicle (HF) morphogenesis is supported by cooperative epithelial–mesenchymal interactions. Previous studies have been focused on the investigation of the crosstalk between HF cell components and, despite wide recognition of its significance, the
e57
mechanism by which surrounding environment supports HF morphogenesis is poorly understood. Recently, the role of dermal fibroblasts in de novo HF regeneration via FGF9 secretion was uncovered in mice. However, whether human dermal fibroblasts are capable of amending the environment to be folliculogenic using similar machinery remains to be dissected. The aim of this study is to assess the ability of human scalp-derived dermal fibroblasts (hsFBs) to modulate their FGF expression profile in response to WNT activation, a key event to initiate HF morphogenesis. In primary culture, hsFBs were distinguished from dermal papilla (DP) and dermal sheath (DS) cells, known trichogenic cell populations, by high expression of FGF5, 13 and 18, potential negative regulators of hair formation and cycling. Interestingly, when exposed to WNT activator CHIR999021, cultured hsFBs suppressed the expression of FGF7, which has been shown to inhibit HF morphogenesis, in a dose dependent manner. WNT activation also up-regulated of FGF1 and 10, two representative DS FGFs, in hsFBs. Importantly, hsDFs tended to increase FGF9 subfamily FGFs, FGF9, 16 and 20 in WNT activated state. Supplementation of FGF9, but not 7, 10 and 16, to human DP cell culture resulted in up-regulation of some DP biomarkers, SRRY4, RGS2 and NOG, implying a role of FGF9 in the maintenance of DP properties. These findings suggested putative contribution of hsFBs to provide folliculogenic microenvironment by modulating regional FGF expression profile responding to WNTs. http://dx.doi.org/10.1016/j.jdermsci.2016.08.177 P09-09 TRPS1 haploinsufficiency results in increased STAT3 and SOX9 mRNA expression in hair follicles in trichorhinophalangeal syndrome Akitaka Shibata ∗ , Kana Tanahashi, Kazumitsu Sugiura, Masashi Akiyama The Department of Dermatology, Nagoya University Graduate School of Medicine, Nagoya, Japan Trichorhinophalangeal syndrome types I and III (TRPS1, OMIM 190350; TRPS3, OMIM 190351) are rare hereditary diseases with autosomal dominant inheritance. Mutations in TRPS1 were reported to cause both TRPS1 and TRPS3. These syndromes have characteristic sparse and slow-growing hair, craniofacial abnormalities such as bulbous pear-shaped nose and skeletal abnormalities. It is known that Trps1 regulates epithelial proliferation in the developing hair follicle via its control of Stat3 and Sox9 expression by the binding to the GATA sequence in mice model. However, the exact function of TRPS1 in the hair follicle has never been clarified yet. We performed mutation analysis of TRPS1 in a Japanese patient with hypotrichosis with skeletal abnormalities, and identified a heterozygous previously unreported mutation c.2191G > T in TRPS1, resulting in an immediate stop codon (p.Glu732X). We confirmed that mRNA decay occurred to the mRNA from the mutant TRPS1 in plucked hairs. The GATAtype zinc finger of TRPS1 protein resident at the position of 896–920 would be lost its repressive function resulted from haploinsufficiency of TRPS1 in the patient. We studied mRNA expression levels of TRPS1-related molecules in the hair follicle of the present TRPS1 patient. We clearly demonstrated for the first time that haploinsufficiency of TRPS1 leads to the up-regulation of STAT3 and SOX9 and that activated STAT3 and catenin 1 signaling might be associated with the hypotrichosis phenotype in TRPS1. Our data give us further insight into the function ofTRPS1 in hair signaling and into the pathomechanisms of TRPS. http://dx.doi.org/10.1016/j.jdermsci.2016.08.178
is also indispensable for maintenance of skin epidermis. To test the hypothesis, HPEK were irradiated with UVB. UVB irradiation stimulated BNIP3 expression and cleavage of caspase3. Surprisingly, suppression of BNIP3 expression induced by UVB irradiation caused a further increase of the cleaved caspase3 protein level, suggesting that BNIP3 has a protective effect against UVB-induced apoptosis in keratinocytes. Overall, our data shed light on functions of BNIP3, an inducer of autophagy, in both differentiation and maintenance of epidermal keratinocytes. http://dx.doi.org/10.1016/j.jdermsci.2016.08.175 P09-07 Plasma cytoid dendritic cells as a key player in the induction of alopecia areata Taisuke Ito ∗ , Takahiro Suzuki, Jun-ichi Sakabe, Atsuko Funakoshi, Yoshiki Tokura Department of Dermatology, Hamamatsu University School of Medicine, Japan Recently, the pathomechanism of alopecia areata (AA) has been thought to be a tissue-specific autoimmune disease. So far, interferon (IFN)-␥ has been regarded as the most important key cytokine that may induce the collapse of HF immune privilege, apoptosis and the upregulation of CXCL10 in C3H/HeJ AA mouse. In this study, we focused on the role of type I IFN, IFN-␣, in the initiation of AA. We previously reported seven cases of AA, including both new onset and recurrent AA, following swine flu viral infection. After viral infection, Th1-mediated immune responses are generally induced by an overproduction of interferons (IFNs). We also showed AA patients just after PEG-IFN-␣2b therapy. In this study, immunohistochemical staining revealed that IFN-␣ producing plasmacytoid dendritic cells (pDCs) densely distributed around HFs in not only lesion skin but also non-lesional skin obtained from C3H/HeJ mice with AA. Flowcytometric analysis showed the increased frequency of pDCs in skin-infiltrated cells of non-lesional skin from C3H/HeJ mice with AA. Real-time PCR also showed higher expression of IFN␣2 and IFN-␣4 mRNA in C3H/HeJ mouse with AA. In vitro study, IFN-␣ inhibited the hair elongation of cultured murine vibrissae in Willium’s E medium by Philpott’s model. In addition, H-2k and CXCL10 expression were upregulated in cultured murine vibrissae by IFN-␣. Imiquimod (IMQ) is the potent inducer of IFN-␣ via TLR7/9 on pDCs. IMQ applied C3H/HeJ mouse showed retardation of hair cycle stage compared to control mouse with infiltration of SiglecH+ pDCs with IFN-␣ expression by immunohistochemical staining. In conclusion, pDCs may play an important role for the initiation of AA by over expression of IFN-␣ via pDCs. http://dx.doi.org/10.1016/j.jdermsci.2016.08.176 P09-08 Human scalp-derived fibroblasts modulate FGF expression profile to be putatively folliculogenic in response to WNT activation Misaki Ise ∗ , Aki Tsukashima, Masayuki Amagai, Manabu Ohyama Department of Dermatology, Keio University School of Medicine, Tokyo, Japan Hair follicle (HF) morphogenesis is supported by cooperative epithelial–mesenchymal interactions. Previous studies have been focused on the investigation of the crosstalk between HF cell components and, despite wide recognition of its significance, the
e57
mechanism by which surrounding environment supports HF morphogenesis is poorly understood. Recently, the role of dermal fibroblasts in de novo HF regeneration via FGF9 secretion was uncovered in mice. However, whether human dermal fibroblasts are capable of amending the environment to be folliculogenic using similar machinery remains to be dissected. The aim of this study is to assess the ability of human scalp-derived dermal fibroblasts (hsFBs) to modulate their FGF expression profile in response to WNT activation, a key event to initiate HF morphogenesis. In primary culture, hsFBs were distinguished from dermal papilla (DP) and dermal sheath (DS) cells, known trichogenic cell populations, by high expression of FGF5, 13 and 18, potential negative regulators of hair formation and cycling. Interestingly, when exposed to WNT activator CHIR999021, cultured hsFBs suppressed the expression of FGF7, which has been shown to inhibit HF morphogenesis, in a dose dependent manner. WNT activation also up-regulated of FGF1 and 10, two representative DS FGFs, in hsFBs. Importantly, hsDFs tended to increase FGF9 subfamily FGFs, FGF9, 16 and 20 in WNT activated state. Supplementation of FGF9, but not 7, 10 and 16, to human DP cell culture resulted in up-regulation of some DP biomarkers, SRRY4, RGS2 and NOG, implying a role of FGF9 in the maintenance of DP properties. These findings suggested putative contribution of hsFBs to provide folliculogenic microenvironment by modulating regional FGF expression profile responding to WNTs. http://dx.doi.org/10.1016/j.jdermsci.2016.08.177 P09-09 TRPS1 haploinsufficiency results in increased STAT3 and SOX9 mRNA expression in hair follicles in trichorhinophalangeal syndrome Akitaka Shibata ∗ , Kana Tanahashi, Kazumitsu Sugiura, Masashi Akiyama The Department of Dermatology, Nagoya University Graduate School of Medicine, Nagoya, Japan Trichorhinophalangeal syndrome types I and III (TRPS1, OMIM 190350; TRPS3, OMIM 190351) are rare hereditary diseases with autosomal dominant inheritance. Mutations in TRPS1 were reported to cause both TRPS1 and TRPS3. These syndromes have characteristic sparse and slow-growing hair, craniofacial abnormalities such as bulbous pear-shaped nose and skeletal abnormalities. It is known that Trps1 regulates epithelial proliferation in the developing hair follicle via its control of Stat3 and Sox9 expression by the binding to the GATA sequence in mice model. However, the exact function of TRPS1 in the hair follicle has never been clarified yet. We performed mutation analysis of TRPS1 in a Japanese patient with hypotrichosis with skeletal abnormalities, and identified a heterozygous previously unreported mutation c.2191G > T in TRPS1, resulting in an immediate stop codon (p.Glu732X). We confirmed that mRNA decay occurred to the mRNA from the mutant TRPS1 in plucked hairs. The GATAtype zinc finger of TRPS1 protein resident at the position of 896–920 would be lost its repressive function resulted from haploinsufficiency of TRPS1 in the patient. We studied mRNA expression levels of TRPS1-related molecules in the hair follicle of the present TRPS1 patient. We clearly demonstrated for the first time that haploinsufficiency of TRPS1 leads to the up-regulation of STAT3 and SOX9 and that activated STAT3 and catenin 1 signaling might be associated with the hypotrichosis phenotype in TRPS1. Our data give us further insight into the function ofTRPS1 in hair signaling and into the pathomechanisms of TRPS. http://dx.doi.org/10.1016/j.jdermsci.2016.08.178