Factor In Melanoma Research Articles

A Conditional Zebrafish MITF Mutation Reveals MITF Levels Are Critical for Melanoma Promotion vs. Regression In Vivo

The microphthalmia-associated transcription factor (MITF) is the “master melanocyte transcription factor” with a complex role in melanoma. MITF protein levels vary between and within clinical specimens, and amplifications and gain- and loss-of-function mutations have been identified in melanoma. How MITF functions in melanoma development and the effects of targeting MITF in vivo are unknown because MITF levels have not been directly tested in a genetic animal model. Here, we use a temperature-sensitive mitf zebrafish mutant to conditionally control endogenous MITF activity. We show that low levels of endogenous MITF activity are oncogenic with BRAFV600E to promote melanoma that reflects the pathology of the human disease. Remarkably, abrogating MITF activity in BRAFV600Emitf melanoma leads to dramatic tumor regression marked by melanophage infiltration and increased apoptosis. These studies are significant because they show that targeting MITF activity is a potent antitumor mechanism, but also show that caution is required because low levels of wild-type MITF activity are oncogenic.

Prevalence of tanning behaviour among young females in Makkah Al Mukarramah

Background: Tanning and exposure to ultraviolet radiation is an important risk factor for melanoma and non-melanoma skin cancer and it is wide practice in developed countries. Aims & Objective: To study the prevalence of tanning behaviour among young female in Makkah, Saudi Arabia and the extent of their awareness of tanning habit. Materials and Methods: A self-administered questionnaire-based cross-sectional survey was carried out from 9 Feb 2013 until 22 June 2013 among 1000 female adolescents aged 14-23 years in Makkah shopping centers, secondary and intermediate female schools and medical and non-medical universities. Results: We found that 20.2 % of respondents reported tanning their skin. Most of our population (89.1%) is willing to stop tanning if the practice proves dangerous for their health. 60.2% of them feel should cut down on their tanning habit and 66.2% have heard that tanning behaviour is dangerous or unhealthy. Conclusion: Tanning use is prevalent among young female in Makkah region. However, they are aware about the dangerous of tanning behaviour.

Impact of Age on the Management of Primary Melanoma Patients

Objectives: Age is an understudied factor when considering treatment options for melanoma. Here, we examine the impact of age on primary melanoma treatment in a prospective cohort of patients. Methods: We used logistic regression models to examine the associations between age and initial treatment, using recurrence and melanoma-specific survival as endpoints. Results: 444 primary melanoma patients were categorized into three groups by age at diagnosis: 19-45 years (24.3%), 46-70 (50.2%), and 71-95 (25.5%). In multivariate models, older patients experienced a higher risk of recurrence (hazard ratio 3.34, 95% confidence interval, CI, 1.53-7.25; p < 0.01). No significant differences were observed in positive biopsy margin rates or extent of surgical margins across age groups. Patients in the middle age group were more likely to receive adjuvant therapy than those in the older group (odds ratio 2.78, 95% CI 1.19-6.45; p = 0.02) and showed a trend to longer disease-free survival when receiving adjuvant therapy (p = 0.09). Conclusion: Our data support age as an independent negative prognostic factor in melanoma. Our data suggest that age does not affect primary surgical treatment but may affect decisions of whether or not patients receive postoperative treatment(s). Further work is needed to better understand the biological variables affecting treatment decisions and efficacy in older patients.

Atopic dermatitis is not a protective factor for melanoma but asthma may be

There is evidence from cohort studies for an inverse association between atopic dermatitis and asthma and cutaneous melanoma. However, these studies have been too heterogeneous and did not show statistically significant results. Also, this association has not been compared to traditional melanoma risk factors. To test for associations between history of atopic disorders and melanoma life-time prevalence, and for associations between atopic disorders and melanoma prognosis. Validated questionnaires from the European Community Respiratory Health Survey and International Study of Asthma and Allergies in Children protocol on life-time prevalence of atopic disorders were sent to 280 patients with histopathologically confirmed melanoma. The control group consisted of their spouses. The skin phototype was also assessed using a validated questionnaire. One hundred and eighty-four melanoma patients and 169 controls responded to the questionnaire. The life-time prevalence of atopic dermatitis and hayfever was not different in melanoma patients (8.7 % vs. 8.2, p = 0.890 and 15.2 vs. 18.3 %, p = 0.432, respectively). Asthma was non-significantly lower in melanoma patients (3.8 vs. 8.2 %, p = 0.075). Atopic melanoma patients did not differ from non-atopic patients in terms of Breslow thickness, metastases and second melanomas. Atopic dermatitis is not a protective factor in cutaneous melanoma but a history of asthma may be.

Personal melanoma risk awareness versus intrinsic risk.

9069 Background: Intrinsic risk factors for melanoma include personal and family history of the condition, a high number of naevi and a light skin phototype (I or II). The objective of this study was to evaluate the correlation between personal awareness of melanoma risk and objective risk factors and to analyze the elements associated with under-or over-evaluation of the actual risk. Methods: EDIFICE melanoma, a nationwide French observational survey, was conducted through phone interviews on a representative sample of 1502 subjects aged ≥ 18 using typical quotas. The survey took place from 28th Sept 2011 to 20th Oct 2011. Results: 393 subjects (26%) had at least one melanoma risk factor: personal: 1%; family history: 11%; high number of naevi: 8% and phototype I-II: 11%. 1109 (74%) had no risk factor. 1029 (73%) had a correct perception of their risk level, 135 (10%) overestimated their risk and 241 (17%) underestimated it. Compared to the control group (correct perception), the population overestimating the melanoma risk is characterised by a higher percentage of individuals living alone (32% vs. 24%, p&lt;0.05), socio-professional category + (38% vs. 28%, p&lt;0.01) and greater alcohol consumption (45% vs. 34%, p&lt;0.02). They are also more likely to expose themselves to the sun (89% vs. 78%, p&lt;0.004) and less likely to use sunscreen protection (58% vs. 44%, p&lt;0.003). A greater proportion of them participates in melanoma screening programmes (21% vs. 14%, p&lt;0.04). The population that underestimates the risk is characterised by lower educational attainment (11% vs. 7%, p&lt;0.05), greater use of high SPF sunscreen (41% vs. 29%, p&lt;0.0004) and a more frequent use of UV sunbeds (9% vs. 6%, p&lt;0.06). Conclusions: Overall, the French have a fair perception of their personal likelihood of developing melanoma. Interestingly, subjects overestimating their intrinsic risk do not behave appropriately with respect to sun protection measures (more sun exposure and less sunscreen protection). On the other hand, subjects underestimating their risk use UV sunbeds more extensively.

The neural crest transcription factor Brn3a is expressed in melanoma and required for cell cycle progression and survival

Pigment cells and neuronal cells both are derived from the neural crest. Here, we describe the Pit-Oct-Unc (POU) domain transcription factor Brn3a, normally involved in neuronal development, to be frequently expressed in melanoma, but not in melanocytes and nevi. RNAi-mediated silencing of Brn3a strongly reduced the viability of melanoma cell lines and decreased tumour growth in vivo. In melanoma cell lines, inhibition of Brn3a caused DNA double-strand breaks as evidenced by Mre11/Rad50-containing nuclear foci. Activated DNA damage signalling caused stabilization of the tumour suppressor p53, which resulted in cell cycle arrest and apoptosis. When Brn3a was ectopically expressed in primary melanocytes and fibroblasts, anchorage-independent growth was increased. In tumourigenic melanocytes and fibroblasts, Brn3a accelerated tumour growth in vivo. Furthermore, Brn3a cooperated with proliferation pathways such as oncogenic BRAF, by reducing oncogene-induced senescence in non-malignant melanocytes. Together, these results identify Brn3a as a new factor in melanoma that is essential for melanoma cell survival and that promotes melanocytic transformation and tumourigenesis.

Lymphatic Invasion Predicts Aggressive Behavior in Melanocytic Tumors of Uncertain Malignant Potential (MELTUMP)

Lymphatic invasion (LI) identified by immunohistochemical (IHC) staining is common in primary cutaneous melanoma, and LI has been shown to be an independent prognostic factor in melanoma. Its prognostic significance in melanocytic tumors of uncertain malignant potential (MELTUMPs) has not been well characterized. This study included 32 patients with provisional diagnoses of MELTUMP. Lesions were evaluated for tumor thickness, the presence of ulceration, mitotic figures, mitotic figures at the base, tumor infiltrating lymphocytes, as well as peritumoral and intratumoral lymphatic density. Dual IHC staining was used to microscopically detect lymphatic endothelium (podoplanin) containing melanoma cells (S100), with the aid of multispectral imaging in select cases. Univariate analysis was performed to identify associations between clinical and pathologic variables and melanoma-related events. The 32 patients had a median follow-up of 111 months. Two patients subsequently died of melanoma-related disease, 1 died of unknown causes, 5 developed nodal metastases, and the remainder showed no evidence of progressive disease. LI was identified in 8/32 patients (25%) by dual IHC staining, which included the 2 patients who died of melanoma-related disease, 1 patient with bulky nodal metastasis, 1/4 patients with microscopic nodal metastases, and 4 patients who showed no evidence of progressive disease. The presence of LI was associated with melanoma metastases or melanoma-related death (P=0.05). The presence of LI by dual IHC in MELTUMPs is associated with a poorer prognosis, specifically with melanoma metastasis, and may therefore serve as a useful prognostic factor for risk stratification of patients with these diagnostically challenging lesions.

Abstract 1516: Melanocyte activation by UVR-induced calprotectin.

Abstract Melanoma is the most common cause of skin cancer death, although it accounts for only 5% of all skin cancers. Epidemiologic and clinical observations have shown that the ultraviolet (UVR) component of sunlight is the most important etiologic factor in melanoma; however, the precise mechanisms by which UVR drives melanomagenesis are unknown. Mutations in oncogenes and tumor suppressor genes have been identified in melanomas, but most of these are not the UVR signature mutations found in non-melanoma skin cancers. This suggests an indirect role for UVR in melanomagenesis, likely as a tumor promoter. UVR is a potent inducer of keratinocyte-derived paracrine and melanocyte/melanoma cell-derived autocrine factors that profoundly affect melanocyte/melanoma cell gene expression and behavior. Many of these mediators are also potent proinflammatory signals that create an inflammatory environment with additional effects on untransformed and transformed melanocytes. We have identified calprotectin, a secreted heterodimer of S100A8 and A9, as one of these mediators. UVR exposure induces S100A8 and A9 transcription in and calprotectin release from human keratinocytes and melanocytes. Human melanocytes and melanoma cells constitutively express TLR4, one of the receptors for calprotectin. Treatment of human melanocytes and melanoma cells with exogenous calprotectin enhances their proliferation, migration, and invasiveness. UVR-induced calprotectin secretion by melanocytes/melanoma cells and surrounding keratinocytes activates melanocytes/melanoma cells and may thus promote melanomagenesis. Citation Format: Kristine von Maltzan, Stephanie H. Shirley, Donna F. Kusewitt. Melanocyte activation by UVR-induced calprotectin. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1516. doi:10.1158/1538-7445.AM2013-1516

Abstract 1365: Sun protection practices among melanoma survivors.

Abstract Purpose: Unprotected sun exposure is a key risk factor for melanoma and further exposure to ultraviolet radiation after a melanoma diagnosis may increase the risk of a subsequent melanoma. We sought to determine whether melanoma survivors observed sun protection practices more vigilantly than the rest of the population. Methods: The National Health Interview Survey (NHIS) is an annual, cross-sectional survey of the civilian, non-institutionalized population of the United States on a wide range of health topics. We used the 2010 NHIS data on self-reported history of melanoma, sun protection practices, and indoor tanning to determine if individuals with a previous history of melanoma adhered more to sun protection practices than the general population. Results: In 2010, the NHIS surveyed 27,120 adults, of whom 171 (0.74%) had a prior history of melanoma. 54.8% of melanoma survivors were men, and 10.2% were under the age of 40. More melanoma survivors reported they always stay in the shade than the general population (15.6% vs. 10.5%, p&amp;lt;0.001); however, 15.4% of melanoma survivors still reported rarely or never staying in the shade. Melanoma survivors were more likely (than their general population counterparts) to always wear a baseball cap/visor (31.3% vs. 18.4%, p=0.028), a wide-brimmed hat (20.5% vs. 6.1%, p&amp;lt;0.001), and a long-sleeved shirt (12.0% vs. 5.2%, p=0.003) when going outside on a warm, sunny day for more than one hour. They were also more likely to report always using sunscreen (32.0% vs. 17.2%, p=0.005); however, 27.3% of melanoma survivors reported never wearing sunscreen when going outside on a warm sunny day for more than one hour. No significant relationship was found between melanoma survivors and the general population in wearing long pants/skirts on a sunny day (22.2% of melanoma survivors always wear such clothing versus 17.7% of the general population, p=0.404). In addition, melanoma survivors were less likely to have used indoor tanning devices in the past 12 months than the general public (2.1% vs. 5.5%, p=0.009). Conclusion: Melanoma survivors, in general, engage in more sun protection practices than the general population. However, the fact that many do not seek shade and never use sunscreen, and some reported tanning bed use, remains a concern for their skin cancer risk. Citation Format: Jeremy Puthumana, Leah Ferrucci, Susan Mayne, Donald Lannin, Anees Chagpar. Sun protection practices among melanoma survivors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1365. doi:10.1158/1538-7445.AM2013-1365

Prognostic value of tumor-infiltrating immune cells in melanoma

Host cells representing an integral component of solid tumors, among them cells contributing to the development of native and adaptive immune responses, can exert both positive and negative effects on the outcome of the disease. Infiltration of T lymphocyte subsets and antigen presenting dendritic cells, crucial in mounting an efficient antitumor immune response, is generally associated with favorable prognosis. On the contrary, accumulation of tumor-associated macrophages, mast cells and neutrophils, playing important roles in chronic inflammatory processes promoting tumor progression, predicts unfavorable disease outcome in most cases. In melanoma, studies on the prognostic impact of the lymphoid infiltrate in general, and that of T cells, yielded controversial results. In our studies, density of activated T lymphocytes correlated with patients' survival, and we obtained similar results in the case of B cells and mature dendritic cells. The amount of both B cells and dendritic cells showed correlation with that of activated T lymphocytes, and their combined analysis identified patient subgroups with different prognosis. According to data from the literature, intense infiltration by neutrophil granulocytes could be associated with shorter survival, while no unambiguous conclusions can be drawn from studies on the prognostic value of tumor-associated macrophages and other immune cell types. Our results indicate that a prominent infiltration of dendritic cells, B cells and activated T lymphocytes can be considered as a favorable prognostic factor in malignant melanoma.

Determinants of sunbed use in a population of Danish adolescents

In Denmark, melanoma is the most common type of cancer in individuals aged 15-34 years. Ultraviolet radiation from sunbeds is a risk factor for melanoma. Knowledge of the characteristics of sunbed users is important in the development and implementation of prevention strategies of sunbed use. The objective of this study was to examine sunbed use and its association with smoking, parental socioeconomic status (SES), friends' attitudes towards artificial tanning, and school environment among adolescents aged 14-18 years at continuation schools in Denmark. We conducted a survey among adolescents in Danish continuation schools in 2011. We examined sunbed use and its association with age, smoking, friends' attitudes towards artificial tanning, parental SES, and shared environment of the continuation school, using logistic regression. Within the past 12 months, 38% of the pupils had used a sunbed (70% girls and 28% boys). There was no difference in sunbed use according to age. Smoking and friends' positive attitudes towards, and higher use of sunbeds were associated with increased use of sunbeds. High SES of mothers' was associated with lower odds for sunbed use among girls. The association of school environment with sunbed use was modest compared with the other variables. Adolescents in continuation schools report a higher use of sunbeds than Danish adolescents as such. Educational interventions should be targeted at preteens, as sunbed use is common in 14-year-olds. Special educational tools are tested in the continuation school environment and may prove effective in this population.

Increased serum level of thymidine kinase 1 correlates with metastatic site in patients with malignant melanoma

Thymidine kinase 1 (TK1) is involved in cancer progression. Metastatic site is a prognostic factor in melanoma. We assessed whether serum levels of TK1 are associated with metastatic sites or prognosis in patients treated for stage IV melanoma. The study included 64 patients, who received dacarbazine or four-drug chemotherapy (dacarbazine, vincristine, bleomycin, and lomustine) both combined with interferon-alfa. Serum samples for TK1 were analyzed by ELISA. The patients (n = 22) with only skin and subcutaneous metastases had significantly lower mean TK1 levels (1,639 pg/ml) than the patients (n = 42) with other distant metastases (2,586 pg/ml, Mann-Whitney, p = 0.031). TK1 levels above the median (1,590 pg/ml) were significantly related to deep lymph node involvement (odds ratios 3.672; 95 % confidence intervals 1.024-12.843, p = 0.036). There were no other significant associations between TK1 levels and tumor burden nor were the levels significantly related to the response to therapy or survival. Those eight patients who had received previous adjuvant IFN-alfa therapy had lower mean TK1 levels (1,735 pg/ml) as compared to those 56 patients without previous IFN-alfa therapy (2,338 pg/ml, analysis of variance, p = 0.026). This is the first study exploring serum TK1 in melanoma. TK1 might be involved in the deep lymphatic dissemination and progression of melanoma metastasis.

TBX2 expression is regulated by PAX3 in the melanocyte lineage.

The paired box homeotic gene 3 (PAX3) is a crucial regulator for the maintenance of melanocytic progenitor cells and has a poorly defined role in melanoma. To understand how PAX3 affects melanocyte and melanoma proliferation, we identified potential PAX3 downstream targets through gene expression profiling. Here, we identify T-box 2 (TBX2), a key developmental regulator of cell identity and an antisenescence factor in melanoma, as a directly regulated PAX3 target. We also found that TBX2 is involved in the survival of melanoma cells and is overexpressed in some melanoma specimens. The identification of TBX2 as a target for PAX3 provides a key insight into how PAX3 may contribute to melanoma evolution and may provide opportunities for prosenescence therapeutic intervention aimed at disrupting the ability of PAX3 to regulate TBX2.

Delivery of interferons and siRNA targeting STAT3 using lentiviral vectors suppresses the growth of murine melanoma

Interferons (IFNs) have been investigated as important cytokines in immunotherapy. The use of IFNs in cancer immunotherapy has had limited success. In this study, IFN genes were delivered into B16 melanoma cells by lentiviral vectors, and their effects on B16 melanoma were comprehensively analyzed. Type II IFN significantly impaired the viability of B16 cells in vitro. Expression of IFNα and IFNβ in B16 cells efficiently suppressed the establishment of inoculated melanoma. However, intratumoral delivery of IFNs alone with lentiviral vectors had no therapeutic effects on established melanoma. To address the lack of response, a lentivector was constructed to simultaneously transfer therapeutic genes and small interfering RNAs (siRNAs). IFNs and siRNA targeting signal transducer and activator of transcription 3 (STAT3), which is a major immune suppressive transcription factor in melanoma, were delivered simultaneously into the tumor milieu. This treatment successfully rescued the response to IFNγ and attenuated the growth of established tumors. This method has the potential to improve the therapeutic effects of IFNs in cancer immunotherapy.

Detection of sentinel lymph nodes by SPECT/CT and planar scintigraphy: The influence of age, gender and BMI

Background : The most important prognostic factor in melanomas, breast cancers and gynecologic tumors is the state of the lymph nodes (LNs). There is not suitable pre-surgery examination procedure of detection of impacted LNs. The use of lymphatic mapping of sentinel lymph node (SLN) helps reduce the morbidity of surgery. Accurate visualization of the SLN is required for the best results. The aim is to assess the role of planar lymphoscintigraphy and fusion imaging of SPECT/CT in SLN detection in patients with various types of tumors and estimation of some factors influencing detection success – age, gender and body mass index (BMI). Methods : Planar scintigraphy and hybrid modality SPECT/CT were performed in 550 patients (pts) (mean age 58.1 ± 13.1 years): 69 pts with gynecological tumors (37 with cervical cancer, 25 pts with endometrial cancer, 7 pts with vulvar carcinoma); in 161 pts with melanomas; and in 318 women and 2 men with breast cancer. The radiopharmaceutical was injected around the tumor, subareolar or around the scar. Planar and SPECT/CT images were interpreted separately by two nuclear medicine physicians. Efficacy of these two techniques to image SLN were compared. We have estimated the influence of the age, gender and BMI on the succes rate of SLN detection. Student's paired t -test was used for comparing numbers of found SLN by both techniques; F-test, Student's t -test, and Fisher's exact test were utilized for age, gender and BMI characteristics comparisons. Values were considered significant when P <0.05. Results : Planar scintigraphy did not image SLN in 77 pts in total (14.0 %); in 8 pts with gynecologic tumors, in 23 pts with melanomas and in 46 pts with breast cancer. SPECT/CT was negative in 49 pts (8.9 %) in total; in 4 pts with gynecologic tumors, in 12 pts with melanomas and in 33 pts with breast cancer. In 199 (36.2 %) pts the number of SLNs captured on SPECT/CT was higher than on planar imaging. 35 foci of uptake (3.1 % from totally visible 1134 foci on planar images) interpreted on planar images as hot LNs were found to be false positive non-nodal sites of uptake when further assessed on SPECT/CT. Influence of the age, gender and BMI: The group of patients with higher number of detected SLN on SPECT/CT than on planar scintigraphy had lower average age than the group of patients with the same number of detected SLN on SPECT/CT and on planar scintigraphy, the difference was statistically significant ( P =0.008). BMI did not differ in these two groups. There has been proportionally higher number of men than women who have had higher detected SLNs on SPECT/CT than on planar scintigrams (difference statistically significant P =0.04). It follows that gender influences the difference of detected SLNs on SPECT/CT and planar scintigraphy in favor of male gender. Conclusion: In some patients with gynecologic, including breast cancers and melanomas SPECT/CT improves detection of SLNs as compared to planar scintigraphy. It can image nodes not visible on planar scintigrams, excludes false positive uptake and it improves anatomical localization of SLNs. We have found the influence of the age and gender on the difference of detected numbers of SLNs by the fusion of SPECT/CT and planar lymphoscintigraphy. This difference was not influenced by BMI.

MC1R variants predisposing to concomitant primary cutaneous melanoma in a monozygotic twin pair

BackgroundConcomitant primary cutaneous melanoma in monozygotic twins has been reported in only two pairs but in neither of them genetic analysis was performed. Two high-penetrance susceptibility genes, CDKN2A and CDK4 and one low-penetrance gene, MC1R, are well-defined genetic risk factors for melanoma. MITF has been recently identified as a novel intermediate risk melanoma-predisposing gene.Case presentationWe describe the extraordinary occurrence of a primary cutaneous invasive melanoma in two 44-year-old identical, female twins, on the same body site within 30 days of each other and report for the first time the genetic analysis of melanoma susceptibility genes in both twins. Data on characteristics of the twins were collected through a standardized questionnaire and skin examination. Exons 1α, 1β, 2 and 3 of CDKN2A, exon 2 of CDK4, the entire open reading frame of MC1R and the recently described MITF c.952 G > A (p.Glu318Lys) variant were investigated by direct sequencing. Sequencing analysis of the high-penetrance susceptibility genes showed no changes in CDKN2A and in exon 2 of the CDK4 gene. Both patients were heterozygous for the same CDKN2A UTR c.*29C > G variant. Interestingly, the same two heterozygous variants of the MC1R were identified in both twins: the c.451C > T (p.Arg151Cys) and the c.456C > A (p.Tyr152*) variants. Neither patient showed the c.952 G > A (p.Glu318Lys) substitution in the MITF gene.ConclusionsIdentification of two high-risk MC1R variants in our identical twins in the absence of CDKN2A and CDK4 mutations highlights the contribution of low penetrance genes, such as MC1R, in melanoma susceptibility.

219P - Assessment Of Circulating Free DNA (CFDNA) and Circulating Melanoma Cells (CMCS) as Prognostic Biomarkers in Metastatic Cutaneous Melanoma

ABSTRACT Background cfDNA is detectable in patients with metastatic cutaneous melanoma and its plasma concentration could potentially predict their clinical outcomes. We assessed the correlation between cfDNA concentration and patients' overall survival and clinical characteristics including known poor prognostic factors in melanoma. Prognostic value of CMCs number and its association with cfDNA level was also explored. Methods Pre-treatment blood samples from 50 patients with metastatic cutaneous melanoma were collected prospectively. cfDNA was extracted from 1 ml of plasma using QIAamp Circulating Nucleic Acids Kit (Qiagen, Hilden, Germany) and quantified by RNase P qPCR (ABI Life Technologies, Foster City, NJ, USA). CMC enumeration per 7.5 ml whole blood was performed within 96 hours of sample collection using the CellSearch Melanoma Kit (Veridex, NJ, USA). Correlation between cfDNA concentration and clinical characteristics and CMC number was performed using χ2-test and Fisher's exact test where appropriate. Survival curves were produced and compared by the Kaplan-Meier method and log-rank test respectively. Results The median concentration of cfDNA was 6.8 ng/ml (range 0-205.8) and median CMC number was 0/7.5ml (range 0-16). Patients with cfDNA concentration of >75% percentile had significantly shorter overall survival compared to those with cfDNA level Conclusions Plasma cfDNA concentration and presence of CMC in 7.5 ml whole blood can predict patient's survival outcomes in metastatic cutaneous melanoma. No association was found between these two biomarkers and this highlights the possibility that they may represent different tumour biological process. Disclosure G. Brady: has an advisory role in Epistem and owns shares in Epistem. G. Clack: employee of Astra-Zeneca pharamceuticals and ownes stock in Astra-zeneca. A. Hughes: Employee of Astra-Zeneca and ownes stock in Astra-Zeneca. All other authors have declared no conflicts of interest.

AIM1 and LINE-1 Epigenetic Aberrations in Tumor and Serum Relate to Melanoma Progression and Disease Outcome

Aberrations in the methylation status of non-coding genomic repeat DNA sequences and specific gene promoter region are important epigenetic events in melanoma progression. Promoter methylation status in LINE-1 and Absent in melanoma-1(AIM1;6q21) associated with melanoma progression and disease outcome was assessed. LINE-1 and AIM1 methylation status was assessed in paraffin-embedded archival tissues(PEAT)(n=133) and melanoma patients’ serum(n=56). LINE-1 U-Index(hypomethylation) and AIM1 were analyzed in microdissected melanoma PEAT sections. The LINE-1 U-Index of melanoma(n=100) was significantly higher than that of normal skin(n=14) and nevi(n=12)(P=0.0004). LINE-1 U-Index level was elevated with increasing AJCC stage(P<0.0001). AIM1 promoter hypermethylation was found in higher frequency(P=0.005) in metastatic melanoma(65%) than in primary melanomas(38%). When analyzed, high LINE-1 U-Index and/or AIM1 methylation in melanomas were associated with disease-free survival(DFS) and overall survival(OS) in Stage I/II patients (P=0.017, 0.027; respectively). In multivariate analysis, melanoma AIM1 methylation status was a significant prognostic factor of OS(P=0.032). Furthermore, serum unmethylated LINE-1 was at higher levels in both stage III(n=20) and stage IV(n=36) patients compared to healthy donors(n=14)(P=0.022). Circulating methylated AIM1 was detected in patients’ serum and was predictive of OS in Stage IV patients (P=0.009). LINE-1 hypomethylation and AIM1 hypermethylation have prognostic utility in both melanoma patients’ tumors and serum.

Prognostic significance of metastatic cascade in melanoma.

e19029 Background: The ability of metastases to spawn subsequent generations of metastatic lesions is controversial, and its prognostic significance is unknown. We hypothesize that melanoma metastases that have spread to draining lymph nodes will have a worse prognosis than those that have not. Methods: One hundred consecutive patients who had undergone pulmonary resection for metastatic melanoma and who had concomitant hilar and/or mediastinal lymph node evaluation available were identified. Patient demographics, tumor characteristics, mediastinal lymph node status, and overall survival were analyzed. Results: 100 patients (71%) were male and the mean age at metastasectomy was 55 years (mean age at diagnosis 47 years.) The average Breslow thickness was 2.43mm (range 0.46 – 20.55). Twenty-one (21%) of the patients had evidence of metastatic disease present in regional lymph nodes during pulmonary resection. The 5-year overall survival for patients with positive mediastinal lymph nodes was 16% versus 36% with node negative disease (p=0.0005). On multivariate analysis, age at pulmonary resection (HR 1.018, 95% CI 1.002-1.035) and regional lymph nodes status (HR 3.203, 95% CI 1.774-5.781) were found to be independent prognostic indicators of 5-year overall survival. Conclusions: Regional lymph node status is an important prognostic factor in patients with pulmonary metastatic melanoma, and regional nodal assessment should be considered during metastasectomy. The metastatic cascade appears to be a strong prognostic factor in melanoma.

Sun exposure profile in the French population: Results of the EDIFICE melanoma survey.

1566 Background: The main environmental risk factor for melanoma is related to UV-exposure. Our objective was to evaluate in a survey the profile of sun exposition in the French population as well as the level of knowledge about UV risk and UV protection. Methods: “Edifice Melanoma” a nationwide observational survey was conducted in France via phone interviews among a representative sample of 1502 subjects aged ≥ 18, using the quota method. The survey took place shortly after the summer (September 28th to October 20th, 2011). Results: From the total population of 1502 subjects, 330 (22%) denied any sun exposition. Among those declaring being exposed to the sun (1172), 62% affirmed using a photoprotection method (clothes, sun protective cream). The opportunities for sun exposure arose during holidays (85%), sport (79%) or professional activities (23%), with an annual average exposure of 113 days. 902 subjects (77%) declared avoiding sun exposure between 12 and 4 PM. The main sociodemographic characteristics of subjects exposed to the sun, compared to non-exposed ones are as followed (p&lt;0,05): men, aged &lt;40, with no children or with children aged &lt;15, from high social and occupational group, with at least a 2-years university degree. The analysis of sociodemographic characteristics of subjects declaring to use sun protective measures are (p&lt;0,05): women from high social and occupational group, with at least a 2-years university degree, with high income levels. We explored additional risk behavior in the population studied. Subjects who declared having sun exposure were more likely (p&lt;0,05) to be : smokers, alcohol users and/or alcoholic, tanning bed users before the age of 30, and have a high number of nevi (&gt;50). On the other hand, prior history of cancer was associated with a non-exposure behavior. Conclusions: These data provide important new information about sun behavior in the French population and might give important clues in the field of public health and education to increase the effectiveness of melanoma primary prevention campaigns. A better knowledge of the target to improve would provide better results for public health benefit.