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Immunomodulatory effects of classical chemotherapeutic agents for melanoma on M2 macrophages
Abstracts from the 41st Annual Meeting / Journal of Dermatological Science 86 (2017) e1–e95
characteristics of 30 samples which were taken by biopsy and surgical resection from 2004 to 2014. HMGB1 staining intensity was scored from 0 (negative) to 5 (strong), according to the percentages of the positive nuclear or cytoplasmic staining areas of the tumor cells. Tumors with a final staining score of over 3 were considered to be higher HMGB1. Kaplan Meier analysis indicated that patients with tumors with high levels of HMGB1 had a significantly shorter survival period than patients with low levels (p < 0.05). This study demonstrated that high levels of expression of HMGB1 in angiosarcoma and indicate a poor prognosis. http://dx.doi.org/10.1016/j.jdermsci.2017.02.065 P02-15[O1-37] Immunomodulatory effects of classical chemotherapeutic agents for melanoma on M2 macrophages Yumi Kambayashi ∗ , Taku Fujimura, Aya Kakizaki, Sadanori Furudate, Setsuya Aiba The Department of Dermatology, Tohoku University Graduate School of Medicine, Japan Together with regulatory T cells (Tregs), tumor-associated macrophages (TAMs) play roles in maintaining the tumor microenvironment. Though DAV combination chemotherapy using dacarbazine (DTIC), nimustine hydrochroride (ACNU) and vincristine (VCR) has been clinically used for the treatment of malignant melanoma in Japan, immunomodulatory effects of them are not fully understood. Since the majority of TAMs are alternatively activated M2 macrophages that favor tumor development, the aim of this study was to elucidate the immunomodulatory effects of DAV reagents on human monocytes derived M2 macrophages. We investigated mRNA expression and protein production of immune checkpoint molecules, PD-L1, and chemokines from CD163+ CD206+ M2 macrophages derived from peripheral blood mononuclear cells to determine the functional effect of DTIC, ACNU or VCR on M2 macrophages. DTIC and VCR significantly decreased the mRNA expression of PD-L1 expression, which is confirmed by flow cytometry analysis. In addition, ACNU and VCR significantly decreased the mRNA expression of CXCL9, and VCR significantly decreased the mRNA expression of CCL17. Moreover, the mRNA expression and the production of CCL22 were significantly decreased by DTIC, which suggested that DTIC might suppress the recruitment of Tregs in the tumor site. Our present report suggests one of the possible mechanisms of DAV combination chemotherapy for melanoma patients. http://dx.doi.org/10.1016/j.jdermsci.2017.02.066 P02-16[O1-38] Smurfs E3 ubiquitin ligases negatively regulate TGF- signaling in keratinocytes Ken Shiraishi 1,∗ , Masamoto Murakami 1 , Mikiko Tohyama 1 , Xiuju Dai 1 , Takeshi Imamura 2 , Koji Sayama 1 1 Department of Dermatology, Ehime University Graduate School of Medicine, Ehime, Japan 2 Department of Molecular Medicine for Pathogenesis, Ehime University Graduate School of Medicine, Ehime, Japan Members of the transforming growth factor- (TGF-) family are multifunctional proteins that regulate cellular responses, including proliferation, migration, differentiation, and apoptosis.
e23
Intracellular signaling of the TGF- family is mainly mediated by Smad proteins. This signaling is regulated by various post-translational modifications, including ubiquitination. Smad ubiquitin regulatory factor 1 (Smurf1) and Smurf2 are HECT (homologous to the E6-accessory protein. C-terminus)-type E3 ubiquitin ligases, which suppress TGF- signaling through the degradation of Smads and TGF- receptors. However, the biological function and regulation of Smurf1 and Smurf2 in human keratinocytes remain to be elucidated. In this study, we demonstrated that TGF- induced Smad2/3 phosphorylation, enhanced Smad7 protein expression, and upregulated Smurf1 and Smurf2 expression in cultured human keratinocytes. Moreover, TGF--induced Smad2/3 phosphorylation and Smad7 protein expression were significantly increased by the inhibition of both Smurf1 and Smurf2. Furthermore, proliferation and migration of human keratinocytes after TGF- stimulation was significantly inhibited by knockdown of both Smurf1 and Smurf2. These results suggest that Smurfs play an important role as regulators of TGF- signaling in human keratinocytes. http://dx.doi.org/10.1016/j.jdermsci.2017.02.067 P02-17[O1-39] Immunological background of mycosis fungoides develops from inflammatory to steady states Sadanori Furudate ∗ , Taku Fujimura, Aya Kakizaki, Setsuya Aiba The Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai, Japan The immunological background of mycosis fungoides (MF) differs with each tumor stage. As we previously reported, in early stage, cancer stroma contains dense deposition of periostin (POSTN), whereas in advanced stage, the expression of IL-4 is prominent instead of POSTN deposition. Since POSTN modulates cell functions, including the production of chemokines related to monocytes/macrophages (CCL2, CCL4, CCL7), and proinflammatory cytokines such as IL-1 and TNF-␣ from keratinocytes and fibroblasts, we hypothesized that immunological background of MF in early stage might be under inflammatory condition and, that of MF in tumor stage might be under steady state. To prove our hypothesis, we investigated chemokine-production and matrix metalloproteinases (MMPs)-deposition as well as CD205+ DCs and Foxp 3+ regulatory T cells (Tregs) in the lesional skin of each MF stage. As we expected, immunohistochemical staining revealed that proinflammatory chemokines (CXCL5, CCL18, CCL22), MMP12 and PD-L1 were prominent in early MF, whereas CD205+ DCs and Foxp3+ Tregs were increased in advanced MF. Our present data suggested that the immunological background of MF might change from inflammatory state to steady state. http://dx.doi.org/10.1016/j.jdermsci.2017.02.068
characteristics of 30 samples which were taken by biopsy and surgical resection from 2004 to 2014. HMGB1 staining intensity was scored from 0 (negative) to 5 (strong), according to the percentages of the positive nuclear or cytoplasmic staining areas of the tumor cells. Tumors with a final staining score of over 3 were considered to be higher HMGB1. Kaplan Meier analysis indicated that patients with tumors with high levels of HMGB1 had a significantly shorter survival period than patients with low levels (p < 0.05). This study demonstrated that high levels of expression of HMGB1 in angiosarcoma and indicate a poor prognosis. http://dx.doi.org/10.1016/j.jdermsci.2017.02.065 P02-15[O1-37] Immunomodulatory effects of classical chemotherapeutic agents for melanoma on M2 macrophages Yumi Kambayashi ∗ , Taku Fujimura, Aya Kakizaki, Sadanori Furudate, Setsuya Aiba The Department of Dermatology, Tohoku University Graduate School of Medicine, Japan Together with regulatory T cells (Tregs), tumor-associated macrophages (TAMs) play roles in maintaining the tumor microenvironment. Though DAV combination chemotherapy using dacarbazine (DTIC), nimustine hydrochroride (ACNU) and vincristine (VCR) has been clinically used for the treatment of malignant melanoma in Japan, immunomodulatory effects of them are not fully understood. Since the majority of TAMs are alternatively activated M2 macrophages that favor tumor development, the aim of this study was to elucidate the immunomodulatory effects of DAV reagents on human monocytes derived M2 macrophages. We investigated mRNA expression and protein production of immune checkpoint molecules, PD-L1, and chemokines from CD163+ CD206+ M2 macrophages derived from peripheral blood mononuclear cells to determine the functional effect of DTIC, ACNU or VCR on M2 macrophages. DTIC and VCR significantly decreased the mRNA expression of PD-L1 expression, which is confirmed by flow cytometry analysis. In addition, ACNU and VCR significantly decreased the mRNA expression of CXCL9, and VCR significantly decreased the mRNA expression of CCL17. Moreover, the mRNA expression and the production of CCL22 were significantly decreased by DTIC, which suggested that DTIC might suppress the recruitment of Tregs in the tumor site. Our present report suggests one of the possible mechanisms of DAV combination chemotherapy for melanoma patients. http://dx.doi.org/10.1016/j.jdermsci.2017.02.066 P02-16[O1-38] Smurfs E3 ubiquitin ligases negatively regulate TGF- signaling in keratinocytes Ken Shiraishi 1,∗ , Masamoto Murakami 1 , Mikiko Tohyama 1 , Xiuju Dai 1 , Takeshi Imamura 2 , Koji Sayama 1 1 Department of Dermatology, Ehime University Graduate School of Medicine, Ehime, Japan 2 Department of Molecular Medicine for Pathogenesis, Ehime University Graduate School of Medicine, Ehime, Japan Members of the transforming growth factor- (TGF-) family are multifunctional proteins that regulate cellular responses, including proliferation, migration, differentiation, and apoptosis.
e23
Intracellular signaling of the TGF- family is mainly mediated by Smad proteins. This signaling is regulated by various post-translational modifications, including ubiquitination. Smad ubiquitin regulatory factor 1 (Smurf1) and Smurf2 are HECT (homologous to the E6-accessory protein. C-terminus)-type E3 ubiquitin ligases, which suppress TGF- signaling through the degradation of Smads and TGF- receptors. However, the biological function and regulation of Smurf1 and Smurf2 in human keratinocytes remain to be elucidated. In this study, we demonstrated that TGF- induced Smad2/3 phosphorylation, enhanced Smad7 protein expression, and upregulated Smurf1 and Smurf2 expression in cultured human keratinocytes. Moreover, TGF--induced Smad2/3 phosphorylation and Smad7 protein expression were significantly increased by the inhibition of both Smurf1 and Smurf2. Furthermore, proliferation and migration of human keratinocytes after TGF- stimulation was significantly inhibited by knockdown of both Smurf1 and Smurf2. These results suggest that Smurfs play an important role as regulators of TGF- signaling in human keratinocytes. http://dx.doi.org/10.1016/j.jdermsci.2017.02.067 P02-17[O1-39] Immunological background of mycosis fungoides develops from inflammatory to steady states Sadanori Furudate ∗ , Taku Fujimura, Aya Kakizaki, Setsuya Aiba The Department of Dermatology, Tohoku University Graduate School of Medicine, Sendai, Japan The immunological background of mycosis fungoides (MF) differs with each tumor stage. As we previously reported, in early stage, cancer stroma contains dense deposition of periostin (POSTN), whereas in advanced stage, the expression of IL-4 is prominent instead of POSTN deposition. Since POSTN modulates cell functions, including the production of chemokines related to monocytes/macrophages (CCL2, CCL4, CCL7), and proinflammatory cytokines such as IL-1 and TNF-␣ from keratinocytes and fibroblasts, we hypothesized that immunological background of MF in early stage might be under inflammatory condition and, that of MF in tumor stage might be under steady state. To prove our hypothesis, we investigated chemokine-production and matrix metalloproteinases (MMPs)-deposition as well as CD205+ DCs and Foxp 3+ regulatory T cells (Tregs) in the lesional skin of each MF stage. As we expected, immunohistochemical staining revealed that proinflammatory chemokines (CXCL5, CCL18, CCL22), MMP12 and PD-L1 were prominent in early MF, whereas CD205+ DCs and Foxp3+ Tregs were increased in advanced MF. Our present data suggested that the immunological background of MF might change from inflammatory state to steady state. http://dx.doi.org/10.1016/j.jdermsci.2017.02.068