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827 Melanoma imaging biomarkers show strong spectral dependance
ABSTRACTS | Pigmentation and Melanoma 825
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Measurement of skin pigmentation using a chromameter in a 3-dimensional epidermal model containing functional melanocytes B Breyfogle, M Bachelor and M Klausner MatTek Corporation, Ashland, MA Various cosmetic or skin care pharmaceutical formulations augment skin pigmentation either for the intended purpose of skin lightening or as an off target drug effect. A convenient way to screen such effects utilizes MelanoDerm, a highly differentiated, three-dimensional tissue culture model of human epidermis containing normal human melanocytes and keratinocytes. Use of this model can provide valuable in vitro data as an early screening tool prior to the commencement of costly clinical trials. In this study, pigmentation was evaluated over the course of 2-3 weeks using a tristimulus chromometer to measure brightness (L*), yellowness (b*) and redness (a*) in MelanoDerm tissue produced with normal human melanocytes from Black, Asian, or Caucasian donors. In parallel to measurements taken with the chromameter, total melanin content of tissues was also quantified. Over time, cultures became increasingly pigmented with retention of normal epithelial morphology with the expected pigmentation level of the donor tissue, i.e. Black>Asian>Caucasian when cultured in media containing alpha-MSH and beta-FGF. Several OTC skin lightening products were also evaluated in cultures containing normal human melanocytes from Black donors. Over the 2-3 week treatment period, control cultures became increasingly pigmented while tissues treated topically with cosmetic skin lightening agents containing tyosinase inhibitors such as kojic acid and magnesium ascorbyl phosphate remained distinctly lighter when compared to control cultures. After 14 days in culture, total melanin content was found to inversely correlate with surface reflectance (L*). The results described herein suggest that this model is useful for evaluating melanogenesis, skin lightening, and other pigmentation phenomena of skin in vitro. In particular, this study highlights two distinct endpoints, total melanin content and skin color measurement that can be used to evaluate skin pigmentation in vitro.
Identification of long non-coding RNAs regulating the development of malignant melanoma M Jingjing1, R Ge2, S Guo3, W Zhang4, T Gao1 and C Li1 1 Department of Dermatology, Xiijing Hospital, Fourth Military Medical University, Xi’an, Shaanxi, China, 2 Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi’an, Shanxi, China, 3 Department of Dermatology, Xijing Hospital, Xi’an, Shaanxi, China and 4 Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi’an, Shaanxi, China Long non-coding RNAs (lncRNAs), as a subgroup of non-coding RNAs, contribute important functions to the progression of a variety of human tumours. However, the clinical significance and biological mechanisms of lncRNAs in the progression of melanoma remain largely unknown. The objective of this study is to examine the expression and functions of lncRNAs in the tumorigenesis and progression of melanoma. LncRNA expression profiling of primary and metastatic melanoma and pigmented nevi was performed to identify tumour-related lncRNAs. Compared with pigmented nevi, there are 1646 lncRNAs with significantly different expression in melanoma. Coexpression networks of protein-coding RNAs and lncRNAs were constructed. The predicted target genes of lncRNAs mainly participate in cancer pathway. Then we focused on the most differently expressed lncRNAs and used quantitative RT-PCR to confirm their expression in melanoma and pigmented nevi. We identified melanoma-associated lncRNA (MAL) as a potential regulator of melanoma progression, which displayed a remarkable trend of increasing expression levels from nevi to carcinoma tissues. Furthermore, functional analysis demonstrated that knock-down the expression of MAL lead to the significant suppression of migration and invasion of melanoma cell lines. Taken together, our results reveal the potential role of lncRNAs in the development of melanoma and MAL maybe an oncogenic lncRNA that promotes tumor development.
Melanoma imaging biomarkers show strong spectral dependance S Leachman1, K Linden2 and JG Krueger3 1 OHSU, Portland, OR, 2 University of California Irvine, Irvine, CA and 3 Rockefeller University, New York, NY Previous data suggest that melanoma imaging biomarkers may have spectral dependence. Here we confirm and extend these observations by use of a novel hyperspectral dermatoscope using 21 wavelengths from 350nm to 950nm. In our cohort of 73 patients imaged with the hyperspectral dermatoscope, we find that melanoma imaging biomarkers have more intense values outside the visible spectrum. Two examples are: (1) the statistical deviation of branch lengths in pigmented network is greater in the near infrared than in the red and (2) image variation due to superficial epidermal atypia is greater in the ultraviolet. Beyond extending our visible imaging biomarkers outside the visible spectrum, completely new imaging biomarkers, such as oxymetric irregularity, that may be linked to metabolic and angiogenic activity, and that can only be achieved in the presence of heavy pigment by using the entire spectrum, show great promise to noninvasively elucidate underlying biology in pigmented neoplasms
Dysbiosis of gut microbiota by ampicillin exacerbates vitiligo S Akhtar1, E Dellacecca1, V Engelhard2, K Knight1 and C Le Poole1 1 Loyola University Chicago, Maywood, IL and 2 University of Virginia, Charlottesville, VA The intestinal tract carries a complex and diverse microbial community which plays an important role in maintaining overall well-being. Disruption of the gut microbiota (or dysbiosis) has been linked to disorders such as obesity and inflammatory bowel disease, but can also drive disease development at ‘distant’ sites, including autoimmunity, infectious disease and cancer. Vitiligo is characterized by T cell mediated destruction of mature epidermal melanocytes, supported by limited availability of Treg. The effects of ampicillin and neomycin on depigmentation were investigated in vitiligo-prone FH-A2D mice. These antibiotics target broad spectrum bacteria or gram negative and anaerobic species, respectively. Nine pregnant FH-A2D mice were randomly assigned to one of three groups and given either plain drinking water, 1.0 mg/ml of ampicillin or 1.0 mg/ml of neomycin in drinking water. Antibiotic treatment continued after the pups were born. Starting at the age of 4 weeks, pups (8-10 per group) were assessed for depigmentation on a biweekly basis. Up to the age of 10 weeks no considerable difference in depigmentation was observed in the three treatment groups. However, starting at 12 weeks of age a sharp increase in percent depigmentation was seen in mice given ampicillin in drinking water, resulting in 4-fold increased depigmentation by 18 weeks. A more subtle increase in depigmentation (1.8-fold) was observed in the neomycin group. Both antibiotics will reduce colonization by gram negative species including B. fragilis. Overall, our data demonstrate that dysbiosis of the gut microbiota exacerbates vitiligo development in mice.
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Fluoxetine promotes human hair follicle pigmentation: A new anti-greying strategy J Gherardini1, J Che´ret1, M Bertolini1 and R Paus2 1 Univ of Mu¨nster, Muenster, NordrheinWestfalen, Germany and 2 Univ of Manchester, Manchester, United Kingdom Besides regulating complex central nervous system functions, the neurotransmitter, serotonin (5-HT), is produced and act in the periphery (e.g. by modulating receptors involved in pain and pruritus). 5-HT itself reportedly promotes pigmentation in vitro (human melanocyte cell lines), and in vivo (mice), and selective serotonin reuptake inhibitors (SSRI), namely Fluoxetine, can stimulate melanin production in murine hair follicles (HFs). Therefore, we asked whether Fluoxetine exerts any effects on human HF pigmentation in organ-cultured scalp anagen VI HFs treated with Fluoxetine (1mM and 100nM) for 48h. Quantitative (immuno-) histomorphometry for standard HF pigmentation parameters revealed that Fluoxetine significantly increased melanin production in the hair matrix (Masson-Fontana (MF) histochemistry) and a-melanocyte-stimulating hormone (a-MSH) protein expression in the outer root sheath (ORS) ex vivo.Next, we asked whether Fluoxetine can even stimulate some degree of re-pigmentation of white HFs over 6 days of HF organ culture. Quantitative (immuno-)histomorphometric analyses for a-MSH, adrenocorticotropic hormone (ACTH), tyrosinase activity in situ did not reveal significant effects. However, white anagen VI HFs revealed upregulated immunoreactivity for corticotropin-releasing hormone (CRH) in the ORS, and for the pre-melanosome protein (gp100) in the matrix of white HFs. Most importantly, actual melanin production in the HF pigmentary unit was also significantly induced in white anagen VI HFs, as assessed by MF. Taken together, these data suggest a new role for Fluoxetine in human skin physiology and pathology: Fluoxetine may not only retard HF depigmentation by stimulating the HF pigmentary unit, but also can promote the re-pigmentation of white HFs. Therefore, topically applied fluoxetine and other SSRIs deserve to be fully explored as candidate anti-hair greying agents.
S142 Journal of Investigative Dermatology (2017), Volume 137
Gene-UV interactions determining sun damage D Lynn1, E Meyer2, J Aalborg1, N Asdigian1, C Little1, T Terzian1, M Berwick3, S Mokrohisky1, J Morelli1, RP Dellavalle4, L Crane1 and N Box1 1 University of Colorado, Aurora, CO, 2 University of Colorado, Denver, CO, 3 University of New Mexico, Albuquerque, NM and 4 U.S. Department of Veterans Affairs, Eastern Colorado Health Care System, Denver, CO Skin cancers that form after ultraviolet (UV) light exposure occur more frequently than any other cancer type, and in the case of melanoma, they are often fatal. UV exposure induces these cancers through DNA mutation, and causes damage such as aging, solar elastosis and hyperpigmentation. Ultraviolet (UV) facial photographs obtained using the Canfield Visia UV camera may detect sub-visible skin damage caused by UV exposure, and could be used to help track and minimize the accumulation of sun damage. Nevertheless, it is not known how these skin damage scores relate to melanoma risk factors such as UV exposure history or to genetic factors that may predispose to their formation. We are currently following a group of 1,145 children with annual skin exams and telephone interviews and collecting a comprehensive longitudinal set of skin cancer risk factor information. We have used facial UV photography to generate a sun damage score on 550 children. Here we report that facial sun damage scores correlate with known skin cancer and melanoma risk factors such as UV exposure history and genetic risk factors such as MC1R, IRF4, and TYR (among others). Thus we show that UV photography may be used to accurately identify, based on genetic makeup, sun damage associated with an individual’s UV exposure history. Moreover, we report those loci that interact with sun exposure history and those that act in an additive fashion to maximize the effects of UV exposure on sun damage. We are working to determine if this technology may be used to identify unique high risk groups that may benefit from dermatologic surveillance and preventive action.
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Measurement of skin pigmentation using a chromameter in a 3-dimensional epidermal model containing functional melanocytes B Breyfogle, M Bachelor and M Klausner MatTek Corporation, Ashland, MA Various cosmetic or skin care pharmaceutical formulations augment skin pigmentation either for the intended purpose of skin lightening or as an off target drug effect. A convenient way to screen such effects utilizes MelanoDerm, a highly differentiated, three-dimensional tissue culture model of human epidermis containing normal human melanocytes and keratinocytes. Use of this model can provide valuable in vitro data as an early screening tool prior to the commencement of costly clinical trials. In this study, pigmentation was evaluated over the course of 2-3 weeks using a tristimulus chromometer to measure brightness (L*), yellowness (b*) and redness (a*) in MelanoDerm tissue produced with normal human melanocytes from Black, Asian, or Caucasian donors. In parallel to measurements taken with the chromameter, total melanin content of tissues was also quantified. Over time, cultures became increasingly pigmented with retention of normal epithelial morphology with the expected pigmentation level of the donor tissue, i.e. Black>Asian>Caucasian when cultured in media containing alpha-MSH and beta-FGF. Several OTC skin lightening products were also evaluated in cultures containing normal human melanocytes from Black donors. Over the 2-3 week treatment period, control cultures became increasingly pigmented while tissues treated topically with cosmetic skin lightening agents containing tyosinase inhibitors such as kojic acid and magnesium ascorbyl phosphate remained distinctly lighter when compared to control cultures. After 14 days in culture, total melanin content was found to inversely correlate with surface reflectance (L*). The results described herein suggest that this model is useful for evaluating melanogenesis, skin lightening, and other pigmentation phenomena of skin in vitro. In particular, this study highlights two distinct endpoints, total melanin content and skin color measurement that can be used to evaluate skin pigmentation in vitro.
Identification of long non-coding RNAs regulating the development of malignant melanoma M Jingjing1, R Ge2, S Guo3, W Zhang4, T Gao1 and C Li1 1 Department of Dermatology, Xiijing Hospital, Fourth Military Medical University, Xi’an, Shaanxi, China, 2 Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi’an, Shanxi, China, 3 Department of Dermatology, Xijing Hospital, Xi’an, Shaanxi, China and 4 Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi’an, Shaanxi, China Long non-coding RNAs (lncRNAs), as a subgroup of non-coding RNAs, contribute important functions to the progression of a variety of human tumours. However, the clinical significance and biological mechanisms of lncRNAs in the progression of melanoma remain largely unknown. The objective of this study is to examine the expression and functions of lncRNAs in the tumorigenesis and progression of melanoma. LncRNA expression profiling of primary and metastatic melanoma and pigmented nevi was performed to identify tumour-related lncRNAs. Compared with pigmented nevi, there are 1646 lncRNAs with significantly different expression in melanoma. Coexpression networks of protein-coding RNAs and lncRNAs were constructed. The predicted target genes of lncRNAs mainly participate in cancer pathway. Then we focused on the most differently expressed lncRNAs and used quantitative RT-PCR to confirm their expression in melanoma and pigmented nevi. We identified melanoma-associated lncRNA (MAL) as a potential regulator of melanoma progression, which displayed a remarkable trend of increasing expression levels from nevi to carcinoma tissues. Furthermore, functional analysis demonstrated that knock-down the expression of MAL lead to the significant suppression of migration and invasion of melanoma cell lines. Taken together, our results reveal the potential role of lncRNAs in the development of melanoma and MAL maybe an oncogenic lncRNA that promotes tumor development.
Melanoma imaging biomarkers show strong spectral dependance S Leachman1, K Linden2 and JG Krueger3 1 OHSU, Portland, OR, 2 University of California Irvine, Irvine, CA and 3 Rockefeller University, New York, NY Previous data suggest that melanoma imaging biomarkers may have spectral dependence. Here we confirm and extend these observations by use of a novel hyperspectral dermatoscope using 21 wavelengths from 350nm to 950nm. In our cohort of 73 patients imaged with the hyperspectral dermatoscope, we find that melanoma imaging biomarkers have more intense values outside the visible spectrum. Two examples are: (1) the statistical deviation of branch lengths in pigmented network is greater in the near infrared than in the red and (2) image variation due to superficial epidermal atypia is greater in the ultraviolet. Beyond extending our visible imaging biomarkers outside the visible spectrum, completely new imaging biomarkers, such as oxymetric irregularity, that may be linked to metabolic and angiogenic activity, and that can only be achieved in the presence of heavy pigment by using the entire spectrum, show great promise to noninvasively elucidate underlying biology in pigmented neoplasms
Dysbiosis of gut microbiota by ampicillin exacerbates vitiligo S Akhtar1, E Dellacecca1, V Engelhard2, K Knight1 and C Le Poole1 1 Loyola University Chicago, Maywood, IL and 2 University of Virginia, Charlottesville, VA The intestinal tract carries a complex and diverse microbial community which plays an important role in maintaining overall well-being. Disruption of the gut microbiota (or dysbiosis) has been linked to disorders such as obesity and inflammatory bowel disease, but can also drive disease development at ‘distant’ sites, including autoimmunity, infectious disease and cancer. Vitiligo is characterized by T cell mediated destruction of mature epidermal melanocytes, supported by limited availability of Treg. The effects of ampicillin and neomycin on depigmentation were investigated in vitiligo-prone FH-A2D mice. These antibiotics target broad spectrum bacteria or gram negative and anaerobic species, respectively. Nine pregnant FH-A2D mice were randomly assigned to one of three groups and given either plain drinking water, 1.0 mg/ml of ampicillin or 1.0 mg/ml of neomycin in drinking water. Antibiotic treatment continued after the pups were born. Starting at the age of 4 weeks, pups (8-10 per group) were assessed for depigmentation on a biweekly basis. Up to the age of 10 weeks no considerable difference in depigmentation was observed in the three treatment groups. However, starting at 12 weeks of age a sharp increase in percent depigmentation was seen in mice given ampicillin in drinking water, resulting in 4-fold increased depigmentation by 18 weeks. A more subtle increase in depigmentation (1.8-fold) was observed in the neomycin group. Both antibiotics will reduce colonization by gram negative species including B. fragilis. Overall, our data demonstrate that dysbiosis of the gut microbiota exacerbates vitiligo development in mice.
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830
Fluoxetine promotes human hair follicle pigmentation: A new anti-greying strategy J Gherardini1, J Che´ret1, M Bertolini1 and R Paus2 1 Univ of Mu¨nster, Muenster, NordrheinWestfalen, Germany and 2 Univ of Manchester, Manchester, United Kingdom Besides regulating complex central nervous system functions, the neurotransmitter, serotonin (5-HT), is produced and act in the periphery (e.g. by modulating receptors involved in pain and pruritus). 5-HT itself reportedly promotes pigmentation in vitro (human melanocyte cell lines), and in vivo (mice), and selective serotonin reuptake inhibitors (SSRI), namely Fluoxetine, can stimulate melanin production in murine hair follicles (HFs). Therefore, we asked whether Fluoxetine exerts any effects on human HF pigmentation in organ-cultured scalp anagen VI HFs treated with Fluoxetine (1mM and 100nM) for 48h. Quantitative (immuno-) histomorphometry for standard HF pigmentation parameters revealed that Fluoxetine significantly increased melanin production in the hair matrix (Masson-Fontana (MF) histochemistry) and a-melanocyte-stimulating hormone (a-MSH) protein expression in the outer root sheath (ORS) ex vivo.Next, we asked whether Fluoxetine can even stimulate some degree of re-pigmentation of white HFs over 6 days of HF organ culture. Quantitative (immuno-)histomorphometric analyses for a-MSH, adrenocorticotropic hormone (ACTH), tyrosinase activity in situ did not reveal significant effects. However, white anagen VI HFs revealed upregulated immunoreactivity for corticotropin-releasing hormone (CRH) in the ORS, and for the pre-melanosome protein (gp100) in the matrix of white HFs. Most importantly, actual melanin production in the HF pigmentary unit was also significantly induced in white anagen VI HFs, as assessed by MF. Taken together, these data suggest a new role for Fluoxetine in human skin physiology and pathology: Fluoxetine may not only retard HF depigmentation by stimulating the HF pigmentary unit, but also can promote the re-pigmentation of white HFs. Therefore, topically applied fluoxetine and other SSRIs deserve to be fully explored as candidate anti-hair greying agents.
S142 Journal of Investigative Dermatology (2017), Volume 137
Gene-UV interactions determining sun damage D Lynn1, E Meyer2, J Aalborg1, N Asdigian1, C Little1, T Terzian1, M Berwick3, S Mokrohisky1, J Morelli1, RP Dellavalle4, L Crane1 and N Box1 1 University of Colorado, Aurora, CO, 2 University of Colorado, Denver, CO, 3 University of New Mexico, Albuquerque, NM and 4 U.S. Department of Veterans Affairs, Eastern Colorado Health Care System, Denver, CO Skin cancers that form after ultraviolet (UV) light exposure occur more frequently than any other cancer type, and in the case of melanoma, they are often fatal. UV exposure induces these cancers through DNA mutation, and causes damage such as aging, solar elastosis and hyperpigmentation. Ultraviolet (UV) facial photographs obtained using the Canfield Visia UV camera may detect sub-visible skin damage caused by UV exposure, and could be used to help track and minimize the accumulation of sun damage. Nevertheless, it is not known how these skin damage scores relate to melanoma risk factors such as UV exposure history or to genetic factors that may predispose to their formation. We are currently following a group of 1,145 children with annual skin exams and telephone interviews and collecting a comprehensive longitudinal set of skin cancer risk factor information. We have used facial UV photography to generate a sun damage score on 550 children. Here we report that facial sun damage scores correlate with known skin cancer and melanoma risk factors such as UV exposure history and genetic risk factors such as MC1R, IRF4, and TYR (among others). Thus we show that UV photography may be used to accurately identify, based on genetic makeup, sun damage associated with an individual’s UV exposure history. Moreover, we report those loci that interact with sun exposure history and those that act in an additive fashion to maximize the effects of UV exposure on sun damage. We are working to determine if this technology may be used to identify unique high risk groups that may benefit from dermatologic surveillance and preventive action.