Non-melanoma Patients Research Articles

SkinViT: A transformer based method for Melanoma and Nonmelanoma classification.

Over the past few decades, skin cancer has emerged as a major global health concern. The efficacy of skin cancer treatment greatly depends upon early diagnosis and effective treatment. The automated classification of Melanoma and Nonmelanoma is quite challenging task due to presence of high visual similarities across different classes and variabilities within each class. According to the best of our knowledge, this study represents the classification of Melanoma and Nonmelanoma utilising Basal Cell Carcinoma (BCC) and Squamous Cell Carcinoma (SCC) under the Nonmelanoma class for the first time. Therefore, this research focuses on automated detection of different skin cancer types to provide assistance to the dermatologists in timely diagnosis and treatment of Melanoma and Nonmelanoma patients. Recently, artificial intelligence (AI) methods have gained popularity where Convolutional Neural Networks (CNNs) are employed to accurately classify various skin diseases. However, CNN has limitation in its ability to capture global contextual information which may lead to missing important information. In order to address this issue, this research explores the outlook attention mechanism inspired by vision outlooker, which improves important features while suppressing noisy features. The proposed SkinViT architecture integrates an outlooker block, transformer block and MLP head block to efficiently capture both fine level and global features in order to enhance the accuracy of Melanoma and Nonmelanoma classification. The proposed SkinViT method is assessed by different performance metrics such as recall, precision, classification accuracy, and F1 score. We performed extensive experiments on three datasets, Dataset1 which is extracted from ISIC2019, Dataset2 collected from various online dermatological database and Dataset3 combines both datasets. The proposed SkinViT achieved 0.9109 accuracy on Dataset1, 0.8911 accuracy on Dataset3 and 0.8611 accuracy on Dataset2. Moreover, the proposed SkinViT method outperformed other SOTA models and displayed higher accuracy compared to the previous work in the literature. The proposed method demonstrated higher performance efficiency in classification of Melanoma and Nonmelanoma dermoscopic images. This work is expected to inspire further research in implementing a system for detecting skin cancer that can assist dermatologists in timely diagnosing Melanoma and Nonmelanoma patients.

Clinical Outcomes Following an Initial Stereotactic Radiosurgery Course for Brain Metastases from Melanoma

Brain metastases (BM) are common in melanoma patients. The effect of gene mutations is not well characterized since first-line metastatic therapy has shifted from chemotherapy (CHT) to molecularly targeted therapies (TT) and immunotherapy (IO). We report outcomes of melanoma BM patients stratified by molecular subtype and pre-stereotactic radiosurgery (SRS) systemic therapy. We identified all patients completing an initial SRS course for BM at two institutions between 1/2015 and 12/2020. Patients who had prior WBRT and/or resection were eligible. Demographic and clinical parameters were collected, along with melanoma tumor molecular characteristics. Intracranial progression (ICP) was defined as any radiographic distant and/or in-field progression per multidisciplinary consensus. Overall survival (OS) and freedom from ICP (FFICP) were estimated via the Kaplan Meier method. From a total of 1383 SRS BM patients, we identified 118 (8.5%) with melanoma. Median follow up was 8.7 months, median age 64 years (IQR 51-72), 81% had cutaneous origin, and 55% had a KPS of 90-100. Molecular subtypes included BRAF (45%), NRAS (9.3%), and c-KIT (3.4%). Overall, 61% received IO prior to SRS, while 25% and 9.3% received TT and CHT prior to SRS respectively. 60% of patients harboring a mutation received IO as first line therapy, 10% received TT, and 30% received both TT and IO prior to SRS. BRAFmut patients more likely to have received TT prior to SRS (43% vs 9.2%, p<0.001) compared to BRAFwt patients. Median OS was 9.7 months (95% CI 7.8-13) and was not significantly different from non-melanoma patients (p = 0.6). Median FFICP was worse for melanoma patients (5.9 mos, 95% CI 3.5-8.5) than non-melanoma patients (8.96 mos, 95% CI 8.2-9.7, p = 0.009). A total of 72 ICP events occurred, with 56 (77.8%) distant ICP cases, 3 (4.2%) in-field ICP, and 13 (18%) ICP events that were radionecrosis (RN) only. RN was associated with the presence of a targetable mutation (18% vs 2%, p = 0.006) and receipt of TT pre-SRS (36% vs 9.8%, p = 0.001). BRAFmut patients had significantly worse FFICP (3.8 mos, 95% CI 3.0-6.8) compared to BRAFwt patients (8.5 mos, 95% CI 5.8-30.2, p = 0.006), although median OS was not significantly different (9.6 mos, 95% CI 6.9-16 vs 10.7 mos, 95% CI 6.7-15.5, p = 0.8). NRASmut was associated with better FFICP (29 mos, 95% CI 2.94-NA, p = 0.02). In this modern, multi-institutional cohort of SRS patients, melanoma BM patients had worse FFICP compared to non-melanoma BM patients, and BRAFmut patients had worse FFICP than BRAFwt patients. RN was associated with mutational status and receipt of TT pre-SRS. OS did not vary significantly across groups. This analysis may help inform systemic therapy decisions and future genomic studies for patients with BMs from melanoma.

Associations of serum carotenoids with the severity of sunburn and the risk of cancer: A cross-sectional analysis of 1999-2018 NHANES data.

Sunburn is a common problem for outdoor workers and casual outdoor walkers. Carotenoids are important elements in normal function of skin tissue and skin metabolism and are critical in the development of some cancers. However, the possible relationships between sunburn sensitivity, carotenoids and the risk of cancers remain unknown. To explore the associations of serum carotenoids with sunburn severity and the risk of cancers. A cross-sectional study from the National Health and Nutrition Examination Survey from 1999 to 2018 were conducted. The relationship between sunburn and serum carotenoids, cancers were investigated by unconditional or ordinal logistic regression. Mediation analysis was used to explore the effect of carotenoids on the relationship between sunburn and cancers. A total of 25,440 US adults from 1999 to 2018 were enrolled in this study. There were significant differences in sex, race and natural hair color between the sunburn and non-sunburn people. The severity of sunburn was significantly associated with serum trans-β-carotene, cis-β-carotene, combined lutein, and vitamin A. The odds ratios of severe reactions were 5.065 (95% CI: 2.266-11.318) in melanoma patients, 5.776 (95% CI: 3.362-9.922) in non-melanoma patients, and 1.880 (95% CI: 1.484-2.380) in non-skin cancers patients. Additionally, serum carotenoids were partially attributable to the effect of sunburn on skin and non-skin cancers. Sunburn severity was associated with cancers, and severer sunburn was related with higher risk of cancers. Serum carotenoids were also associated with sunburn severity. Moreover, the relationship between sunburn and cancers was mediated by some serum carotenoids.

33597 Checkpoint inhibitor–induced vitiligo in nonmelanoma patients

Checkpoint inhibitors (ICIs) are anticancer therapies with a well-known association of vitiligo induction in melanoma patients. ICI-induced vitiligo is typically rapidly progressive and associated with increased tumor response, possibly because it indicates the body’s heightened immune response to melanocytes. Previously, ICI-induced vitiligo in nonmelanoma cancers had only been described in scattered case reports. Here we describe a cohort of melanoma and nonmelanoma patients with ICI-induced vitiligo. This is a retrospective cohort study from a single institution’s electronic medical record for cancer patients treated with CPIs who subsequently developed vitiligo. We identified 151 patients with ICI-induced vitiligo, 19 (12.6%) nonmelanoma and 132 (77.4%) melanoma patients. Time to onset of vitiligo was nearly doubled in the nonmelanoma cohort, however, this is confounded by possible delayed diagnosis or underreporting of this asymptomatic condition in patients who do not regularly receive skin exams. The majority of patients had a stable course of vitiligo with 91.4% receiving no treatment in this largely Caucasian cohort. Two patients with nonmelanoma cancers and Fitzpatrick type IV or above skin received treatment with narrowband ultraviolet B light therapy and topical steroids with near-complete response. This study highlights the occurrence of CPI-induced vitiligo in a variety of nonmelanoma cancers, where skin of color patients will be more prevalent and the need for treatment will potentially be more urgent. Further study is needed to elucidate the mechanism of CPI-induced vitiligo and determine if nonmelanoma cancers have the same association between vitiligo and increased tumor response.

Immune checkpoint inhibitors\u2013related encephalitis in melanoma and non-melanoma cancer patients: a single center experience

BackgroundTreatment with immune checkpoint inhibitors (ICI) has greatly improved survival for patients with a number of malignant diseases in recent years. Neurological immune-related adverse events (n-irAE) of varying severity have been reported in the literature. We aimed to identify the incidence of n-irAE, focusing on immune-related encephalitis (IRE), in patients treated with ICI for multiple non-hematological malignancies in our institution.MethodsAll patients with histologically verified cancer that received treatment with ICI at the Sheba Medical Center between January 2017 and August 2019 were surveyed. Medical records for each patient were reviewed and information regarding n-irAE was recorded.ResultsIn total, 1993 patients were included. Eleven cases of IRE were recorded, affecting 0.55% of patients overall, eight had non-melanoma cancer. Eight patients had made a full recovery.ConclusionsIRE is a n-irAE more frequent than previously reported, particularly in non-melanoma patients. The diagnostic criteria and optimal treatment needs to be determined. ICI re-challenge after IRE can be considered for selected patients.

1022MO Clinical potential of adoptive cell therapy with tumour infiltrating lymphocytes therapy in combination with checkpoint inhibitors in non-melanoma patients

1022MO Clinical potential of adoptive cell therapy with tumour infiltrating lymphocytes therapy in combination with checkpoint inhibitors in non-melanoma patients

Immune-Related Adverse Events and Outcomes in Patients with Advanced Non–Small Cell Lung Cancer: A Predictive Marker of Efficacy?

Immune-Related Adverse Events and Outcomes in Patients with Advanced Non–Small Cell Lung Cancer: A Predictive Marker of Efficacy?

Abstract 5393: BRAF mutant allele fraction in circulating tumor DNA as marker of treatment response in BRAF mutated non malignant melanoma cancers identified in the Copenhagen Prospective Personalized Oncology study

Abstract Background: BRAF inhibitors have been approved in BRAF mutated malignant melanoma. However, in other BRAF mutated cancers, the effect of BRAF inhibition has been studied less extensively. BRAF mutated non-melanoma cancer patients (pts) were identified in a prospective genomic profiling study. The effect of targeted treatment was assessed by BRAF mutant allele fraction (AF) in circulating tumor DNA (ctDNA) and radiological response. Methods: Pts had tumor biopsies performed at enrolment and, if feasible, on or after treatment. Biopsies were subjected to whole exome sequencing, RNA sequencing and SNP array. Blood samples for ctDNA analysis were collected at baseline and longitudinally during treatment. Mutant BRAF fragments were quantified using droplet digital PCR (ddPCR) or by targeted next generation sequencing (NGS). Radiological response was evaluated according to RECIST 1.1. Results: Twenty-two BRAF mutated non-melanoma cancer pts (lung n=2, colorectal n=14, bile duct n=4, prostate n=1, pancreas n=1) were identified out of a total of 405 (5 %) pts enrolled in the Copenhagen Prospective Personalized Oncology (CoPPO) study. In all tumors we identified the BRAF V600E mutation except in one prostate cancer pt (BRAF K601E). Fifteen pts were treated with dabrafenib/trametinib (lung, bile duct), vemurafenib/panitumumab +/- irinotecan (colorectal) or vemurafenib (pancreas, prostate). Two pts were non-evaluable because one patient requested early termination and one has pending tumor evaluation. All pts had tumor reduction, except one patient with colon cancer. Six out of 13 (46%) pts achieved partial response (lung n=2, bile duct n=1, colorectal n= 3) and 7/13 (44%) had stable disease (prostate n=1, bile duct n=1, colorectal n= 5) as best response according to RECIST 1.1. Median progression-free survival was 20 weeks. Eighteen pts analysed by ddPCR or NGS at baseline had detectable mutant BRAF in the plasma obtained before initiation of therapy. Eight of the 18 pts had ctDNA assessed during treatment and all presented with a reduction in mutant BRAF AF corresponding to a reduction in tumor volume. Moreover, in 5/8 pts, time of best radiological response coincided with the lowest detectable BRAF mutant AF. Analysis of on- and post treatment biopsies and matched ctDNA samples are pending to identify putative mechanisms of resistance and will be reported at the meeting. Conclusion: Targeted therapy seems active in BRAF mutated non-melanoma cancer pts and treatment response is evaluable by monitoring mutant BRAF in ctDNA during therapy. Citation Format: Lise Barlebo Ahlborn, Ida Viller Tuxen, Olga Oestrup, Ane Yde Schmidt, Cecilia Brunhoff Håkansson, Finn Celius Nielsen, Ulrik Lassen, Christina Westmose Yde, Morten Mau-Sorensen. BRAF mutant allele fraction in circulating tumor DNA as marker of treatment response in BRAF mutated non malignant melanoma cancers identified in the Copenhagen Prospective Personalized Oncology study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5393. doi:10.1158/1538-7445.AM2017-5393

Immune-Related Adverse Events as a Biomarker in Non-Melanoma Patients Treated with Programmed Cell Death 1 Inhibitors.

The programmed death 1 (PD-1) checkpoint inhibitors (CKIs) can lead to immune-related adverse events (irAEs). We sought to evaluate whether the development of irAEs correlates with treatment response in non-melanoma malignancies. We conducted a retrospective study of patients who received anti-PD-1 CKI monotherapy at Fox Chase Cancer Center. Endpoints included overall response rate (ORR), time to next therapy or death (TTNTD), and overall survival (OS). Fisher's exact tests and logistic regression models were used to determine the association between irAE incidence and ORR, and Kaplan-Meier curves with log-rank tests and Cox regression models were used for the comparison of TTNTD and OS. Between November 2011 and November 2016, 160 patients were treated with >1 dose of an anti-PD-1 CKI. Seventy-three (46%) were treated on a clinical trial. Immune-related adverse events were noted in 64 patients (40%), with steroids required in 36 (23%). Of the 142 patients evaluable for clinical response, 28 patients (20%) achieved a partial response at first scan. An association between irAEs and ORR was seen in clinical trial patients (p = .007), but not in non-trial patients (p = .13). When controlling for clinical trial participation and cancer type using multivariate analysis, low-grade irAEs had higher ORR (p = .017) and longer TTNTD (p = .008). No association between irAE incidence and OS was seen (p = .827). Immune-related adverse events that required steroid treatment were marginally associated with increased TTNTD (p = .05, hazard ratio 0.62) but were not associated with OS (p = .13). We demonstrate several positive associations between the development of irAEs and clinical outcomes in non-melanoma patients treated with PD-1 CKIs, for which further validation is required. This study evaluated whether the development of immune-related adverse events in non-melanoma patients treated with programmed cell death 1 checkpoint inhibitors correlates with improved clinical outcomes. The results indicate that for a subset of patients, in particular those with low-grade immune-related adverse events, immune-related adverse events predicted for an improved response rate and longer time to next therapy or death.

Patient centered outcomes for skin cancer treatment: A single day Delphi process to assess the importance of treatment themes to a representative panel of skin cancer patients.

e21079 Background: Increased emphasis is being placed on patient centered outcomes. There is little existing literature on the patient population's prioritization of outcomes in skin cancer care. A PCORI-supported Patient Outcomes Summit was organized to address this. Methods: Patients were recruited from patient support groups, solicitation of ASDS member practices, as well as personal recruitment of patients from the practices of the investigators. A one-day Patient Outcome Summit was convened in Arlington, VA in August 2016. Patients attendees were given orientation to existing outcomes for skin cancer as well as the Delphi process. They then participated in a two round Delphi process by run non-physician facilitators (AB and EA). Group discussion was used to solicit outcomes for inclusion in the rating rounds. Outcomes were then rated anonymously on a 1 (lowest relevance) to 9 (highest relevance) scale. Patients were then able to view the anonymous feedback of the group and rerate the items as they deemed fit based on the group feedback. Results: 21 patients participated in the Delphi process (5 men and 21 women). 10 were melanoma and 11 were non-melanoma patients. 56 treatment related outcomes and themes were identified. The ten most highly rated themes are shown in the table. Conclusions: The first ever skin cancer patient outcome summit was convened to address the lack of patient-centered data in the skin cancer literature and guide measure development. Some traditional outcomes (recurrence) were identifiend. However, the majority of the themes rated most important by the patient attendees center around inclusion of the patient in the decision making process and the physician-patient communication. Increased emphasis and education in these domains may drive increased patient satisfaction in skin cancer care. [Table: see text]

Cost-effectiveness analysis in melanoma detection: A transition model applied to dermoscopy

AimThe main aim of this study is to demonstrate how our melanoma disease model (MDM) can be used for cost-effectiveness analyses (CEAs) in the melanoma detection field. In particular, we used the data of two cohorts of Belgian melanoma patients to investigate the cost-effectiveness of dermoscopy. MethodsA MDM, previously constructed to calculate the melanoma burden, was slightly modified to be suitable for CEAs. Two cohorts of patients entered into the model to calculate morbidity, mortality and costs. These cohorts were constituted by melanoma patients diagnosed by dermatologists adequately, or not adequately, trained in dermoscopy. Effectiveness and costs were calculated for each cohort and compared. Effectiveness was expressed in quality-adjusted life years (QALYs), a composite measure depending on melanoma-related morbidity and mortality. Costs included costs of treatment and follow-up as well as costs of detection in non-melanoma patients and costs of excision and pathology of benign lesions excised to rule out melanoma. ResultsThe result of our analysis concluded that melanoma diagnosis by dermatologists adequately trained in dermoscopy resulted in both a gain of QALYs (less morbidity and/or mortality) and a reduction in costs. ConclusionThis study demonstrates how our MDM can be used in CEAs in the melanoma detection field. The model and the methodology suggested in this paper were applied to two cohorts of Belgian melanoma patients. Their analysis concluded that adequate dermoscopy training is cost-effective. The results should be confirmed by a large-scale randomised study.

Dynamics of mutant BRAF V600E in free circulating DNA (fcDNA) of non-melanoma cancer patients (pts) in response to treatment with BRAF and MEK/EGFR inhibitors.

11531Background: Therapies targeting mutant BRAF V600E have changed practice in the treatment of metastatic melanoma and are currently tested in other malignancies. Treatment response can be monito...

Immune related adverse events as a biomarker in non-melanoma patients treated with programmed death 1 inhibitors (PD-1Is).

3069Background: The PD-1Is pembrolizumab (P) and nivolumab (N) have demonstrated clinical activity in multiple tumor types. They can lead to unique immune related adverse events (irAEs). Studies ha...

Association of eosinophila with efficacy in non-melanoma patients treated with programmed death 1 inhibitors.

e14510Background: Programmed death-1 inhibitors (PD-1Is) have demonstrated durable responses in patients (pts) with a variety of tumor types. We explored the association between eosinophilia and re...

Quantitative Measurement of Melanoma Spread in Sentinel Lymph Nodes and Survival

Sentinel lymph node spread is a crucial factor in melanoma outcome. We aimed to define the impact of minimal cancer spread and of increasing numbers of disseminated cancer cells on melanoma-specific survival. We analyzed 1,834 sentinel nodes from 1,027 patients with ultrasound node-negative melanoma who underwent sentinel node biopsy between February 8, 2000, and June 19, 2008, by histopathology including immunohistochemistry and quantitative immunocytology. For immunocytology we recorded the number of disseminated cancer cells (DCCs) per million lymph node cells (DCC density [DCCD]) after disaggregation and immunostaining for the melanocytic marker gp100. None of the control lymph nodes from non-melanoma patients (n = 52) harbored gp100-positive cells. We analyzed gp100-positive cells from melanoma patients by comparative genomic hybridization and found, in 45 of 46 patients tested, gp100-positive cells displaying genomic alterations. At a median follow-up of 49 mo (range 3-123 mo), 138 patients (13.4%) had died from melanoma. Increased DCCD was associated with increased risk for death due to melanoma (univariable analysis; p<0.001; hazard ratio 1.81, 95% CI 1.61-2.01, for a 10-fold increase in DCCD + 1). Even patients with a positive DCCD ≤3 had an increased risk of dying from melanoma compared to patients with DCCD = 0 (p = 0.04; hazard ratio 1.63, 95% CI 1.02-2.58). Upon multivariable testing DCCD was a stronger predictor of death than histopathology. The final model included thickness, DCCD, and ulceration (all p<0.001) as the most relevant prognostic factors, was internally validated by bootstrapping, and provided superior survival prediction compared to the current American Joint Committee on Cancer staging categories. Cancer cell dissemination to the sentinel node is a quantitative risk factor for melanoma death. A model based on the combined quantitative effects of DCCD, tumor thickness, and ulceration predicted outcome best, particularly at longer follow-up. If these results are validated in an independent study, establishing quantitative immunocytology in histopathological laboratories may be useful clinically.

Compliance with follow-up among patients with melanoma and non-melanoma skin cancers

A history of cutaneous malignancies puts patients at an increased risk of developing additional skin cancers, but there is little data available regarding compliance with any recommended follow-up regimens. To report on compliance with physician recommended follow-up regimens among patients diagnosed with melanoma and non-melanoma skin cancers. 105 patients with cutaneous melanoma and 1151 patients with non-melanoma skin cancers diagnosed between December 1996 and December 2001 were identified through biopsy records. The records of all identified patients were then retrospectively reviewed for compliance with physician recommended follow-up regimens over a 60-month period. At 60 months following initial diagnosis, 22.6% of melanoma patients and 19.3% of non-melanoma patients were still continuing follow-up examinations. However, only 10.5% of melanoma patients and 7.2% of non-melanoma patients were compliant with the recommended follow-up schedule. Among melanoma patients, Breslow thickness correlated positively with duration of follow-up (P = 0.03). The frequency of additional primary non-melanoma and melanoma skin cancers was positively correlated with duration of follow-up among patients with non-melanoma skin cancers (P ≤ 0.001). Patient compliance with physician recommended follow-up regimens is generally poor; further research and intervention is necessary to identify and address the underlying causes.

The New AJCC Guidelines in Practice

The American Joint Commission on Cancer 2010 guideline changes recommend all patients with single-cell melanoma metastasis identified by immunhistochemical staining of sentinel lymph nodes to be classified as N1 stage. In this study we evaluated the utility of adding microphthalmia transcription factor (MITF) to our current sentinel node evaluation protocol. Twenty benign lymph nodes from nonmelanoma patients were stained with MITF, MART-1/Melan-A, and HMB-45. Hematoxylin and eosin-stained and immunohistochemically stained slides were assessed for the frequency of positive staining, pattern and location of staining, and for the presence of pigment within the node. Fifteen of 20 (75%) lymph nodes evaluated showed >1 cell staining with MITF in variable locations and patterns throughout the lymph node. MART-1/Melan-A-positive cells were identified in 4 of 20 cases (20.0%), mostly in the sinusoidal location (3/4). Fifty percent (10/20) of the lymph nodes contained cells that stained positive for HMB-45. Nine of 10 of the HMB-45-positive cells were in a sinusoidal location corresponding to easily discernible pigmented histiocytes. We conclude that MITF stains nonmelanocytic cells in benign lymph nodes 75% of the time. MITF has a much higher rate of positive staining in benign lymph nodes compared with MART-1/Melan-A, which had positive staining only 20% of the time. MITF showed 2 different staining patterns, type A and type B. Both patterns could be difficult to distinguish from true metastasis, and thus adding MITF to sentinel node evaluation panels is not helpful and may be histologically vexing.

Expression of miR-16 is not a suitable reference for analysis of serum microRNAs in melanoma patients

Background: The evolving paradigm shift towards the molecular characterization of melanoma has expanded to include studies of microRNA (miRNA) expression. As miR-16 has been utilized as a normalizer in serum-based miRNA studies in several cancers, we evaluated miR-16 expression as a potential reference for normalization of serum miRNA expression in melanoma patients. Methods: 143 primary cutaneous melanoma patients who presented to New York University (NYU) Langone Medical Center for surgical resection of AJCC stage I-III disease were studied. In addition, sera samples from 60 control subjects were utilized including 22 healthy volunteers, 13 rheumatoid arthritis patients, 20 non-melanoma cancer patients (10 renal cell carcinoma and 10 bladder cancer), and 5 Atypical Mole Syndrome patients. The Kruskal-Wallis test (k = 6) or Wilcoxon test (k = 2) with Bonferroni correction was used for analyses of miR-16 expression in melanoma patients compared to various control groups, using raw Ct values directly. The Kruskal-Wallis test was used to compare miR-16 expression across stages of melanoma. The equivalence test for independent samples was used to test the equivalence of miR-16 expression among different groups. Results: No significant differential expression of miR-16 was observed between melanoma patients and healthy volunteers (Wilcoxon test, p = 0.37). However, miR-16 did show a significant difference in expression as it related to stage of melanoma (p = 0.015). Additionally, the equivalence test was unable to confirm equivalent expression of miR-16 in any melanoma versus control group pair. Conclusion: Our data indicate that miR-16 cannot be used as a universal normalizer in sera studies of melanoma patients.

Immuno-Expression of Human Melanoma Stem Cell Markers in Tissues at Different Stages of the Disease

Human cutaneous melanoma can be one of the most aggressive tumors and is extremely resistant to all current therapeutic modalities. Immunohistochemistry (IHC) studies were undertaken to assess independent relative frequency on preferential overexpression and simultaneous expression of four stem cell markers; CD133+, ABCB5+, CD166, and Nestin, at different stages of the disease. Five-micron sections from paraffin blocks from primary melanoma, lymph node (LN) metastases, distant metastases, benign nevi from non-melanoma patients (NMP), and patients with past history of melanoma (MP) were IHC stained with monoclonal antibodies (mAb) to the four stem cell markers. Overexpression of CD133+ was noted in tissues from LN and distant metastases compared to benign nevi (P < 0.0022, P < 0.013, respectively). Overexpression of ABCB5+ was observed comparing primary melanoma, LN, and distant metastases to benign nevi from NMP (P < 0.0063, P < 0.001, P < 0.00058, respectively). Significant overexpression of ABCB5+ was noted in tissues from LN and distant metastases compared with benign nevi from MP (P < 0.0003, P < 0.0068). None of the benign nevi of NMP demonstrated ABCB5+. This study clearly shows the existence of a distinct hyperpolarized population of stem cells and may implicate genetic factors in human cutaneous melanoma. Simultaneous overexpression of CD133+ and Nestin could reflect on their origin or lineage association with a neural stem cell. These findings may open new perspectives for therapeutic implication of a melanoma stem cell in specific cancer therapy.

The state of systemic immune dysfunction in patients with metastatic melanoma

9067 Background: Immunotherapeutic strategies are often used for the treatment of patients with advanced melanoma. Although successful in pre-clinical models, clinical efficacy remains limited. One explanation for this discrepancy is the state of the host’s immune system (immunocompetent mouse vs immunocompromised cancer patient). We’ve previously reported that repeated peptide immunization of patients with metastatic melanoma lead to peptide-specific tolerance, not active immunity, which correlated with tumor progression. Further studies revealed that plasma from patients with advanced melanoma inhibited normal dendritic cell function in vitro, indicating the existence of a soluble mediator of immune dysfunction. These observations lead to our current effort to understand the differences in immune homeostasis in patients across stages of melanoma. Methods: Immune homeostasis was ascertained by analysis of relative frequencies of NK, dendritic and T-cells present in peripheral blood, plasma levels of 27 cytokines, and the functional state of antigen specific T-cells in 85 patients with benign and/or dysplastic nevi, 72 patients with stage I- III melanoma and 32 with stage IV melanoma. Matched expression array analysis of tumor biopsies from 19 of the 32 stage IV melanoma patients was performed as well. The data was analyzed using analysis covariance, adjusting for age, gender, and batch with logarithmic or square root transformation of parameters to achieve normality. Results: Notably, the cytokine levels for the Th2 cytokines, IL-4 (78 vs 4.4pg/ml), IL-10 (14 vs 3.8pg/ml), and IL-13 (13 vs 2.3pg/ml), in tumor bearing (stage IV) versus early stage (benign nevi-stage III) patients were significantly higher (p-value<0.02). Matched expression microarray analysis revealed that tumor cells transcribe immunologically relevant molecules that could contribute to the observed Th2 dominant state of systemic chronic inflammation. Conclusions: Tumor bearing, unlike tumor free or non-melanoma patients, appear to exhibit a state of systemic Th2 dominance (chronic inflammation) potentially explaining some of the observed immune dysfunction and poor clinical outcomes of immunotherapeutic interventions in patients with stage IV melanoma. No significant financial relationships to disclose.