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466 Sebaceous carcinoma treated with Mohs micrographic surgery
ABSTRACTS | Melanoma and Other Skin Cancers 465
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Ultraviolet (UV)-A irradiation induces melanoma invasion via enhanced Warburg effect T Baban1, Y Kamenisch2, W Schuller1, A von Thaler1, T Sinnberg1, J Bauer1, G Metzler1, C Garbe1, M Ro¨cken1 and M Berneburg2 1 Department of Dermatology, Eberhard Karls University, Tu¨bingen, Germany and 2 Department of Dermatology, University Regensburg, Regensburg, Germany Melanoma is a malignant tumor for which exposure to ultraviolet (UV) radiation is considered to be an important risk factor. It was recently shown that UVA radiation (320-400nm) is capable to induce murine melanoma, but the role of UVA in the progression of melanoma is still not investigated. During early progression of melanomas before metastasising, most melanomas show initial proliferation and a metabolic characteristic of most proliferating tumor cells is the preference of aerobic glycolysis instead of oxidative phosphorylation (Warburg effect). Here we investigated the role of repetitive UVA radiation in progression of melanoma, especially induction of progression markers, changes in Warburg effect and invasive potential. In skin reconstructs treated with repetitive UVA irradiation initial melanoma cells show increased initial dermal invasion. Upon UVA radiation, initial melanoma cells show increased Warburg effect with increased glucose consumption and increased lactate production. The tumor marker transketolase and phosphorylated Akt kinase, which are involved in metabolic changes and associated with proliferation, are also elevated upon UVA radiation. With in vitro invasion assays we show, that lactate, which is produced via UVA enhanced Warburg effect, increases invasiveness of initial melanoma cells. The expression of tumor relevant matrix metalloproteinases (MMP) and urokinase type plasminogen activator (uPA) are highly upregulated upon UVA irradiation or treatment with lactic acid and MMP and uPA mainly facilitate in vitro invasion. Furthermore treatment with lactic acid alone can mimic the UVA induced increase of the invasive potential in vitro. Therefore we could show in melanoma cells, derived from melanomas of early progression that production of lactate, induced by UVA radiation, increases invasiveness of initial melanoma cells via expression of MMPs.
Sebaceous carcinoma treated with Mohs micrographic surgery J Kim, J Seo and K Chung Dermatology, Yonsei University College of Medicine, Seoul, Korea (the Republic of) Sebaceous carcinoma is a rare, aggressive tumor of sebaceous glands commonly located on the head and neck. Clinically it resembles benign inflammatory condition, which may contribute to higher rates of distant metastasis mortality due to diagnostic delay. Due to its rare occurrence, optimal treatment modality has yet to be established. Earlier, surgical resection with wide local excision was performed for most of the cases. However, high recurrence rate is reported after conventional resection. Recent studies have shown Mohs micrographic surgery (MMS), which enables accurate margin control, can lead to less chance of recurrence with more appreciable cosmetic and functional results. The results of three patients with sebaceous carcinoma treated by MMS have been analyzed. Medical records were retrospectively reviewed for the analysis of clinical characteristics and detailed surgical data. Review of literature for treatment modalities and recurrence rates of sebaceous carcinoma was also performed. We have experienced three elderly patients (1 male, 2 female; mean age 82.3) with sebaceous carcinoma. The lesions were located on the nose, forehead and scalp. All three patients were successfully treated with MMS. During the average follow up of 28 months, there was no recurrence of the tumor or associated morbidity. Based on our experience, MMS is a relatively simple and effective treatment for patients for sebaceous carcinoma with low recurrence and patient morbidity. Despite the limitation of being a retrospective study with relatively small sample size, the results of this study show the possibility of Mohs micrographic surgery for rare malignant tumors.
Rare sarcomas of the skin: A case series J Kim, J Seo and K Chung Dermatology, Yonsei University College of Medicine, Seoul, Korea (the Republic of) All sarcomas of deep soft tissues may also arise in superficial locations and there can be important clinicopathologic differences. Recently, we have experienced some kinds of these rare malignancies occurring in the skin. We report herein the four cases of different sarcomas having a primary cutaneous location: Ewing sarcoma, histiocytic sarcoma, myxofibrosarcoma, and liposarcoma. Extensive metastatic work-up, including a total body computed tomographic and positron emission scan (PET/CT scan), showed the neoplasm to be a primary sarcoma of the skin. All four patients presented with negative sentinel lymph node biopsy specimen and were treated by Mohs micrographic surgery (MMS) using paraffinembedded sections, i.e. slow Mohs. We hope that our experience will broaden dermatologists’ understanding of these rare sarcomas arising from the skin.
Estrogen- and progesterone receptor expression in pregnancy-associated melanoma M Fa´bia´n1, P Balogh2, T Krena´cs2, F Rencz4, V Brodszky4, J Ha´rsing1, K Ne´meth3 and S Ka´rpa´ti1 1 Dermatology, Venereology and Dermatooncology, Semmelweis University, Budapest, Hungary, 2 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary, 3 Medical College of Wisconsin, Milwaukee, WI and 4 Health Economics, Corvinus University, Budapest, Hungary Beside breast, ovarian and prostate cancer, part of the scientific community also considers malignant melanoma (MM) as a hormone dependent malignancy, while the role of estrogen receptors (ERs) and progesterone receptor (PR) in MM development and prognosis is still controversial. Pregnancy-associated melanoma (PAM) is diagnosed during gravidity or within one year after delivery (postpartum melanoma). The database of the Dermatology, Venereology and Dermatooncology Clinic of the Semmelweis University (DVDSE) was retrospectively reviewed and all the diagnosed PAM cases during a 12 year period were analyzed in this study. The expression of ERa, ERb and PR was examined by immunohistochemistry (IHC) and the stained slides were subjected to whole slide digitalization using Panoramic Scanner (3DHISTECH). The IHC was evaluated on 81 MM samples (38 PAM and 43 controls (nonpregnant women and men (NPAM)) who underwent surgery for the primary tumor at the DVDSE.We detected cytoplasmic ERb positivity of the tumor cells together with an intensive nuclear positivity in a subpopulation of inflammatory cells. The protein expression of ERb was significantly higher in PAM than in NPAM group. Conversely, the studied tissue samples remained negative for both ERa and PR. Our data also demonstrated that cytoplasmic ERb positivity was associated with lower Breslow thickness and lower mitotic rate, as well as better disease free and overall survival, indicating a positive prognostic role of the receptor expression in melanoma. In summary, the elevated ERb expression in MM may suggest a better outcome of melanomas, however, further investigation in larger patient cohorts and functional studies are still needed to elucidate these questions in detail.
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Experimental T cells immunotherapy by tumor-loaded ECP-generated dendritic antigen presenting cells A Ventura2, A Vassall2, E Robinson2, M Bosenberg2, L Bianchi1, S Chimenti1 and R Edelson2 1 Dermatology, University of Rome Tor Vergata, Rome, Italy and 2 Dermatology, Yale University, New Haven, CT Extracorporeal Photopheresis (ECP) is a widely used immunotherapy for cutaneous T cell lymphoma. We hypothesized that ECP’s physiologic induction of large-scale monocyte-todendritic antigen presenting cell (APC) conversion is mechanistically responsible for its anticancer effect. To investigate this possibility we developed an ECP protocol that is scalable from mouse to man and tested its capacity to produce APCs that, when advantageously tuned and tumor specific antigen-loaded, can limit the growth of otherwise lethal tumors in the engineered Yale University Mouse Melanoma (YUMM) 1.7 model (driven by PTEN loss, BRAFV600E activation and CDKN2A mutations) and in the Murine Colon Adenocarcinoma (MC38). Untreated control mouse tumors were 7 to 10-fold (p<0.001) larger than ECP treated tumors at the endpoint. Depletion of ECP-generated APCs from the cellular vaccine preparation prevented the immunoprotective effect (p<0.01). Depletion of ECP induced APC from the cellular vaccine preparation prevented the immunoprotective effect (p<0.01), indicating a primary role for ECP-induced APC.Depletion of platelets from the ECP-processed cellular suspension also prevented the immunizing effect (p<0.01), substantiating prior in vitro evidence that physiologic ECP-induction of functional APC is signaled by transient monocyte adherence to platelets. When YUMM1.7 tumor bearing mice received unrelated apoptotic MC38 tumor cells as source of the antigen there was statistically no difference than untreated control mice and contrary significant delay seen with the YUMM1.7. This anti-tumor effect is immune mediated and transferrable to naı¨ve C57BL/6 mice receiving splenocytes from mECP-treated mice (p<0.05). Collectively, these results show that, in ECP, platelet-induced tumor-loaded mature APCs are immunotherapeutic for established murine solid tumor. These findings verify a pivotal role for ECP-manufactured APC and suggest the intriguing possibility that ECP can be modified for immunotherapy of solid tumors.
S240 Journal of Investigative Dermatology (2016), Volume 136
Putative driver mutations of Kaposiform hemangioendothelioma detected by exome sequence analysis S Egashira, M Jinnin, M Harada, S Masuguchi, S Fukushima and H Ihn Kumamoto University, Kumamoto city, Japan Kaposiform hemangioendothelioma (KHE) is a rare intermediate malignant vascular tumor. The etiology of the tumor is unknown, and there are no specific treatments. The aim of this study is to identify putative candidates for the responsible mutations by exome sequencing. The genomic DNA for exome sequencing was obtained from the tumor tissue as well as from matched normal tissue of the same KHE patient. The exome sequence was performed on HiSeq2000 sequencer platform. Compared with the reference human genome, there were 80,062 nucleotide changes in the normal tissue, and 73,878 changes in the tumor tissue. Among them, 68,342 changes were common to both tissues. Among oncogenes, germline missense single nucleotide variants (SNVs) were found in TP53 and APC gene in both tumor and normal tissue. As tumor-specific somatic mutations, we observed 81 candidate genes with 4 nonsense changes, 68 missense changes or 9 insertions/deletions. Among them, for example, ITGB2, IL-32 and DIDO1 has been well implicated in the mechanism of angiogenesis or the pathogenesis of malignant disorders. This is a pilot study, and further analysis with increased number of patients is needed to clarify the detailed pathogenesis of this tumor, novel diagnostic methods by detecting specific mutation, and new therapeutic strategies targeting the mutation.
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Ultraviolet (UV)-A irradiation induces melanoma invasion via enhanced Warburg effect T Baban1, Y Kamenisch2, W Schuller1, A von Thaler1, T Sinnberg1, J Bauer1, G Metzler1, C Garbe1, M Ro¨cken1 and M Berneburg2 1 Department of Dermatology, Eberhard Karls University, Tu¨bingen, Germany and 2 Department of Dermatology, University Regensburg, Regensburg, Germany Melanoma is a malignant tumor for which exposure to ultraviolet (UV) radiation is considered to be an important risk factor. It was recently shown that UVA radiation (320-400nm) is capable to induce murine melanoma, but the role of UVA in the progression of melanoma is still not investigated. During early progression of melanomas before metastasising, most melanomas show initial proliferation and a metabolic characteristic of most proliferating tumor cells is the preference of aerobic glycolysis instead of oxidative phosphorylation (Warburg effect). Here we investigated the role of repetitive UVA radiation in progression of melanoma, especially induction of progression markers, changes in Warburg effect and invasive potential. In skin reconstructs treated with repetitive UVA irradiation initial melanoma cells show increased initial dermal invasion. Upon UVA radiation, initial melanoma cells show increased Warburg effect with increased glucose consumption and increased lactate production. The tumor marker transketolase and phosphorylated Akt kinase, which are involved in metabolic changes and associated with proliferation, are also elevated upon UVA radiation. With in vitro invasion assays we show, that lactate, which is produced via UVA enhanced Warburg effect, increases invasiveness of initial melanoma cells. The expression of tumor relevant matrix metalloproteinases (MMP) and urokinase type plasminogen activator (uPA) are highly upregulated upon UVA irradiation or treatment with lactic acid and MMP and uPA mainly facilitate in vitro invasion. Furthermore treatment with lactic acid alone can mimic the UVA induced increase of the invasive potential in vitro. Therefore we could show in melanoma cells, derived from melanomas of early progression that production of lactate, induced by UVA radiation, increases invasiveness of initial melanoma cells via expression of MMPs.
Sebaceous carcinoma treated with Mohs micrographic surgery J Kim, J Seo and K Chung Dermatology, Yonsei University College of Medicine, Seoul, Korea (the Republic of) Sebaceous carcinoma is a rare, aggressive tumor of sebaceous glands commonly located on the head and neck. Clinically it resembles benign inflammatory condition, which may contribute to higher rates of distant metastasis mortality due to diagnostic delay. Due to its rare occurrence, optimal treatment modality has yet to be established. Earlier, surgical resection with wide local excision was performed for most of the cases. However, high recurrence rate is reported after conventional resection. Recent studies have shown Mohs micrographic surgery (MMS), which enables accurate margin control, can lead to less chance of recurrence with more appreciable cosmetic and functional results. The results of three patients with sebaceous carcinoma treated by MMS have been analyzed. Medical records were retrospectively reviewed for the analysis of clinical characteristics and detailed surgical data. Review of literature for treatment modalities and recurrence rates of sebaceous carcinoma was also performed. We have experienced three elderly patients (1 male, 2 female; mean age 82.3) with sebaceous carcinoma. The lesions were located on the nose, forehead and scalp. All three patients were successfully treated with MMS. During the average follow up of 28 months, there was no recurrence of the tumor or associated morbidity. Based on our experience, MMS is a relatively simple and effective treatment for patients for sebaceous carcinoma with low recurrence and patient morbidity. Despite the limitation of being a retrospective study with relatively small sample size, the results of this study show the possibility of Mohs micrographic surgery for rare malignant tumors.
Rare sarcomas of the skin: A case series J Kim, J Seo and K Chung Dermatology, Yonsei University College of Medicine, Seoul, Korea (the Republic of) All sarcomas of deep soft tissues may also arise in superficial locations and there can be important clinicopathologic differences. Recently, we have experienced some kinds of these rare malignancies occurring in the skin. We report herein the four cases of different sarcomas having a primary cutaneous location: Ewing sarcoma, histiocytic sarcoma, myxofibrosarcoma, and liposarcoma. Extensive metastatic work-up, including a total body computed tomographic and positron emission scan (PET/CT scan), showed the neoplasm to be a primary sarcoma of the skin. All four patients presented with negative sentinel lymph node biopsy specimen and were treated by Mohs micrographic surgery (MMS) using paraffinembedded sections, i.e. slow Mohs. We hope that our experience will broaden dermatologists’ understanding of these rare sarcomas arising from the skin.
Estrogen- and progesterone receptor expression in pregnancy-associated melanoma M Fa´bia´n1, P Balogh2, T Krena´cs2, F Rencz4, V Brodszky4, J Ha´rsing1, K Ne´meth3 and S Ka´rpa´ti1 1 Dermatology, Venereology and Dermatooncology, Semmelweis University, Budapest, Hungary, 2 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary, 3 Medical College of Wisconsin, Milwaukee, WI and 4 Health Economics, Corvinus University, Budapest, Hungary Beside breast, ovarian and prostate cancer, part of the scientific community also considers malignant melanoma (MM) as a hormone dependent malignancy, while the role of estrogen receptors (ERs) and progesterone receptor (PR) in MM development and prognosis is still controversial. Pregnancy-associated melanoma (PAM) is diagnosed during gravidity or within one year after delivery (postpartum melanoma). The database of the Dermatology, Venereology and Dermatooncology Clinic of the Semmelweis University (DVDSE) was retrospectively reviewed and all the diagnosed PAM cases during a 12 year period were analyzed in this study. The expression of ERa, ERb and PR was examined by immunohistochemistry (IHC) and the stained slides were subjected to whole slide digitalization using Panoramic Scanner (3DHISTECH). The IHC was evaluated on 81 MM samples (38 PAM and 43 controls (nonpregnant women and men (NPAM)) who underwent surgery for the primary tumor at the DVDSE.We detected cytoplasmic ERb positivity of the tumor cells together with an intensive nuclear positivity in a subpopulation of inflammatory cells. The protein expression of ERb was significantly higher in PAM than in NPAM group. Conversely, the studied tissue samples remained negative for both ERa and PR. Our data also demonstrated that cytoplasmic ERb positivity was associated with lower Breslow thickness and lower mitotic rate, as well as better disease free and overall survival, indicating a positive prognostic role of the receptor expression in melanoma. In summary, the elevated ERb expression in MM may suggest a better outcome of melanomas, however, further investigation in larger patient cohorts and functional studies are still needed to elucidate these questions in detail.
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Experimental T cells immunotherapy by tumor-loaded ECP-generated dendritic antigen presenting cells A Ventura2, A Vassall2, E Robinson2, M Bosenberg2, L Bianchi1, S Chimenti1 and R Edelson2 1 Dermatology, University of Rome Tor Vergata, Rome, Italy and 2 Dermatology, Yale University, New Haven, CT Extracorporeal Photopheresis (ECP) is a widely used immunotherapy for cutaneous T cell lymphoma. We hypothesized that ECP’s physiologic induction of large-scale monocyte-todendritic antigen presenting cell (APC) conversion is mechanistically responsible for its anticancer effect. To investigate this possibility we developed an ECP protocol that is scalable from mouse to man and tested its capacity to produce APCs that, when advantageously tuned and tumor specific antigen-loaded, can limit the growth of otherwise lethal tumors in the engineered Yale University Mouse Melanoma (YUMM) 1.7 model (driven by PTEN loss, BRAFV600E activation and CDKN2A mutations) and in the Murine Colon Adenocarcinoma (MC38). Untreated control mouse tumors were 7 to 10-fold (p<0.001) larger than ECP treated tumors at the endpoint. Depletion of ECP-generated APCs from the cellular vaccine preparation prevented the immunoprotective effect (p<0.01). Depletion of ECP induced APC from the cellular vaccine preparation prevented the immunoprotective effect (p<0.01), indicating a primary role for ECP-induced APC.Depletion of platelets from the ECP-processed cellular suspension also prevented the immunizing effect (p<0.01), substantiating prior in vitro evidence that physiologic ECP-induction of functional APC is signaled by transient monocyte adherence to platelets. When YUMM1.7 tumor bearing mice received unrelated apoptotic MC38 tumor cells as source of the antigen there was statistically no difference than untreated control mice and contrary significant delay seen with the YUMM1.7. This anti-tumor effect is immune mediated and transferrable to naı¨ve C57BL/6 mice receiving splenocytes from mECP-treated mice (p<0.05). Collectively, these results show that, in ECP, platelet-induced tumor-loaded mature APCs are immunotherapeutic for established murine solid tumor. These findings verify a pivotal role for ECP-manufactured APC and suggest the intriguing possibility that ECP can be modified for immunotherapy of solid tumors.
S240 Journal of Investigative Dermatology (2016), Volume 136
Putative driver mutations of Kaposiform hemangioendothelioma detected by exome sequence analysis S Egashira, M Jinnin, M Harada, S Masuguchi, S Fukushima and H Ihn Kumamoto University, Kumamoto city, Japan Kaposiform hemangioendothelioma (KHE) is a rare intermediate malignant vascular tumor. The etiology of the tumor is unknown, and there are no specific treatments. The aim of this study is to identify putative candidates for the responsible mutations by exome sequencing. The genomic DNA for exome sequencing was obtained from the tumor tissue as well as from matched normal tissue of the same KHE patient. The exome sequence was performed on HiSeq2000 sequencer platform. Compared with the reference human genome, there were 80,062 nucleotide changes in the normal tissue, and 73,878 changes in the tumor tissue. Among them, 68,342 changes were common to both tissues. Among oncogenes, germline missense single nucleotide variants (SNVs) were found in TP53 and APC gene in both tumor and normal tissue. As tumor-specific somatic mutations, we observed 81 candidate genes with 4 nonsense changes, 68 missense changes or 9 insertions/deletions. Among them, for example, ITGB2, IL-32 and DIDO1 has been well implicated in the mechanism of angiogenesis or the pathogenesis of malignant disorders. This is a pilot study, and further analysis with increased number of patients is needed to clarify the detailed pathogenesis of this tumor, novel diagnostic methods by detecting specific mutation, and new therapeutic strategies targeting the mutation.