Early-stage Melanoma Research Articles

Impact of adjuvant immunotherapy in resected stage II/III/IV melanoma: A single center experience.

e21565 Background: The incidence of malignant melanoma (MM) in Mediterranean countries is estimated at 3- 5/100.000 inhabitants/year with a predicted rising trend. MM is responsible for over 90% skin cancer deaths due to its tendency to metastasize early. Surgical excision is the treatment of choice for early-stage melanoma, however, 40-60% of patients with high recurrence risk melanoma, thick ulcerated primary melanomas (Stage IIC) and those with lymph node involvement (Stage III), experience a tumor relapse. Adjuvant therapy in resected high-risk melanomas has shown improved Relapse-Free-Survival (RFS), although no Overall Survival (OS) benefit has been observed to date. This retrospective observational study, conducted in one center, aims to evaluate effectiveness and safety of adjuvant therapies for resected high-risk melanoma. Methods: Single center cross-sectional study that included patients with a resected high-risk melanoma candidate for adjuvant treatment, at 2015-2022 period. Convenience sampling 48 patients recruited. Median follow up 55,4 months. Results: Median age was 54y, 52% were male and 93’6 is cutaneous melanoma. 53% were BRAF WT. Median Breslow (mm) was 3’85, median num/mm2 mitosis are 4 and 62’5% had ulceration. 15% presented satelosis or intransit metastasis and 55% had a lymphadenectomy. Melanoma stage: 9%IIB, 11%IIC, 11%IIIA, 25%IIIB, 38%IIIC, 2% IIID and 2% IV. Adjuvant treatment: 22% nivolumab, 20% ipilimumab+nivolumab, 12% ipilimumab, 16% interferon.75% of patients completed adjuvant treatment, 14% stopped for toxicity and 9% presented disease relapse (21% loca, 64% systemic). Best treatment response in relapse was 12% partial response, 37% progression. Toxicity: 18% presented grad ¾ toxicity. Efficacy: median RFS was 78’8m and median OS was NR. Conclusions: Adjuvant treatment is associated with an increased DFI in resected high risk melanoma patients. Adjuvant therapy was generally well tolerated, with manageable adverse events, consistent in type and incidence to those seen in clinical trials that allowed the approval for these compounds in adjuvant and metastatic scenarios. Prospective studies are needed to determine new biomarkers for the selection of those patients who could benefit to a greater extent from adjuvant therapies in resected high-risk melanoma.

Longitudinal circulating tumor DNA monitoring for detection of molecular residual disease in patients with surgically resected stage II/III melanoma.

9582 Background: Over the past decade, advances in immunotherapy (IO) have dramatically improved overall survival (OS) for patients with metastatic melanoma. Immune checkpoint inhibitors and BRAF/MEK inhibitors also became standard of care for surgically resected stage III melanoma, and most recently pembrolizumab was approved for stage IIB/C patients. Although the phase III trials that led to their respective approvals met their primary endpoints for relapse-free survival, no benefit in OS has been observed. Given that a large proportion of early-stage melanoma patients are cured with surgery alone, the decision to proceed with adjuvant therapy should be weighed against the risk of potential life-altering toxicity. There is a high unmet need to develop biomarkers to accurately select patients who are most likely to benefit from adjuvant IO while sparing a group of low risk patients from unnecessary treatment. In this study, we evaluated the prognostic value of personalized, tumor-informed circulating tumor DNA (ctDNA) testing in patients with resectable stage II/III melanoma. Methods: In this real-world study, longitudinal plasma samples (n = 159) were analyzed in real-time from 45 stage IIA-IIID cutaneous melanoma patients treated at Rush University between 03/30/2021 and 01/12/2023. A personalized, tumor-informed ctDNA assay (Signatera bespoke, mPCR-NGS assay) was used for detection and quantification of ctDNA in plasma samples. Results: Personalized, tumor-informed ctDNA assays were successfully designed for 93% (42/45) of patients using tissue samples from either surgical resection (14/42, 33%) or biopsy (26/42, 67%; 16 shave, 10 excisional, 1 punch, 1 core). ctDNA was detectable at one or more time points in 36% (15/42) of patients. We observed a higher ctDNA-positivity rate in stage III patients (11/20, 55%) compared to stage II patients (4/22, 18%). Twelve of the 15 ctDNA-positive patients received adjuvant IO/treatment escalation after a positive ctDNA result; 6 patients had sustained ctDNA clearance, 2 had transient clearance, and 4 continue to receive adjuvant IO and follow-up ctDNA testing. Radiographic recurrence was observed in 7/42 patients, 6 of whom were ctDNA-positive. Ninety-six percent (26/27) of patients with persistent ctDNA-negativity remain clinically and radiographically progression-free. Conclusions: Our data suggest that molecular residual disease detection using a personalized, tumor-informed ctDNA assay is highly prognostic in melanoma patients undergoing curative resection. Sustained clearance of ctDNA was achieved with adjuvant therapy in a subset of this cohort. Prospective studies evaluating the utility of ctDNA to aid clinical decision-making regarding adjuvant treatment intensification or de-escalation are warranted.

Association of biomarkers (BMs) with efficacy of adjuvant nivolumab (NIVO) vs placebo (PBO) in patients with resected stage IIB/C melanoma (CA209-76K).

9504 Background: Adjuvant NIVO significantly prolonged recurrence-free survival (RFS) over PBO in stage IIB/C melanoma in the randomized, phase III Checkmate 76K trial (HR; 95% CI: 0.42 [0.30–0.59]). For the first time, select BMs and their association with RFS were analyzed in early-stage melanoma treated with an anti-PD-1 agent. Methods: Baseline primary tumor and serum BMs were analyzed, including interferon gamma-related gene expression signature (IFNγ-sig), TMB, BRAF mutation, % intratumoral CD8+ T cells, tumor cell PD-L1 expression, and serum CRP levels. BMs were evaluated as continuous variables (high vs low levels) and dichotomized by median or prespecified cutoffs. HRs were estimated using a Cox proportional hazards model. Results: Analysis of continuous BM levels within treatment arms showed that higher IFNγ-sig, TMB, and % CD8+ T cells, and lower CRP levels were associated with prolonged RFS with NIVO but not PBO. The same trend was observed in the dichotomized analyses. No clear association between PD-L1 expression and RFS was identified in either arm. Benefit of NIVO over PBO was seen across all BM subgroups, including by BRAFV600 mutation status. Patients with higher IFNγ-sig, TMB, and % CD8+ T cells, and lower CRP levels showed numerically greater relative benefit with NIVO, although CIs overlapped (Table). Conclusions: Higher IFNγ-sig, TMB, and % CD8+ T cells, and lower CRP levels were associated with longer RFS with NIVO treatment. No BMs assessed were prognostic for RFS in the PBO arm, in contrast to previous reports in melanoma. Prolonged RFS was seen in patients treated with NIVO over PBO across levels of BMs, and irrespective of BRAF mutational status. Composite analyses of BMs with clinical and histopathologic factors will be evaluated for their potential to optimize risk:benefit stratification for NIVO in this patient population. Clinical trial information: NCT04099251 . [Table: see text]

Skin Lesions Detection using Deep Learning Techniques

Abstract: The main category of cancer is skin cancer, which is manifested by certain skin diseases. They have been constrained into various typologies stationed on morphological features, color, structure, and texture. Due to certain factors, the study equips analysis efforts to advance algorithms with greater closeness and tensility in detecting early-stage melanoma. As per the ACS report, melanoma is one of the recurrent cancers in the world. In 2017, around 87,110 new cases were diagnosed in the United States alone. Dermoscope images contain imperfections such as shadows, artifacts, and noise that degrade image essence. To overthrow these objection, deep learning neural networks have been used to detect skin cancer by tampering images. In the proposed work, automatic classification of skin lesions is proposed. Image classification, object detection etc are some of the computer vision tasks in deep CNN which have driven DCNNs to be reliable on several substitute assumptions, tiling its way for new exploration areas. CNN attained performance equivalent to all experts tested, achieving an exact competence equivalent to dermatologists treating skin cancer. In this article, we attempt to improve the Deep Convolutional Neural Networks example using the ImageNet dataset with HAM10000, fortuitously classifying seven categories of skin lesions. HAM10000 is a dataset of 10000 dermoscope images. Layers are fine-tuned applying separate approach such as InceptionV3, InceptionResNet, DenseNet and VGG-16. Over previous years, the power of deep learning-based approaches has enhanced fiercely and their work come out to outperform common image processing approach on classification tasks. However, these categories of machine learning-based accession have important drawbacks. Training requires thousands of annotated images for each class. The idea is to use deep learning algorithms and available dataset assets to bear models with higher accuracy and best results.

Melanoma Skin Cancer Classification based on CNN Deep Learning Algorithms

Melanoma, the deadliest form of skin cancer, is on the rise. The goal of this study is to present a deep learning system implementation for the detection of melanoma lesions on a server equipped with a graphics processing unit (GPU). When applied by a dermatologist, the recommended method might aid in the early detection of this kind of skin cancer. Evidence shows that deep learning may be used in a variety of settings to successfully extract patterns from data such as signals and images. This research presents a convolution neural network–based strategy for identifying early-stage melanoma skin cancer. Images are input into a deep learning model known as a convolutional neural network (CNN) that has already been pre-trained. The CNN classifier, which is trained with large amounts of data, can discriminate between malignant and nonmalignant melanoma. The method's success in the lab bodes well for its potential to aid dermatologists in the early detection of melanoma. However, the experimental results show that the proposed technique excels beyond the state-of-the-art methods in terms of diagnostic accuracy.

Early Stage Melanoma of the Vermillion with Mucosal Melanoma in Situ: A Clinical Conundrum

BackgroundMelanoma is a predominantly cutaneous malignancy associated with sun exposure. Mucosal melanoma is rare and carries a distinct pathogenesis from cutaneous tumors. The vermillion of the lip is a unique site that divides cutaneous from mucosal tissues. Tumors arising on the dry aspect are classified as cutaneous and those of the wet aspect are mucosal. The distinction is importation in tumor staging as all mucosal melanomas are classified as T3-T4b under the current 8th edition American Joint Committee of Cancer (AJCC) guidelines.MethodsWe describe a case of early stage melanoma of the vermillion with associated mucosal melanoma in situ. We discuss nuances of management at this site and the distinctions between cutaneous versus mucosal melanomas with a review of the literature.ResultsOur patient was treated surgically with 2-3 cm margins. Residual melanoma in situ was present at the mucosal margin on final pathology which required a second surgery for margin revision. The case was discussed at tumor board with recommendation for no further treatment.ConclusionsThe nuances between the vermillion and mucosal lip must be understood for appropriate staging and treatment of melanomas. The paucity of literature on melanomas affecting this site render management decisions challenging. Multidisciplinary discussion is essential for guiding care.

Transcriptome Analysis Identifies Oncogenic Tissue Remodeling during Progression from Common Nevi to Early Melanoma

Early detection and treatment of melanoma, the most aggressive skin cancer, improves the median 5-year survival rate of patients from 25% to 99%. Melanoma development involves a stepwise process during which genetic changes drive histological alterations within nevi and surrounding tissue. Here, we perform a comprehensive analysis of publicly available gene expression datasets of melanoma, common or congenital nevi (CN), and dysplastic nevi (DN), and assess molecular & genetic pathways leading to early melanoma. Our results demonstrate several pathways reflective of ongoing local structural tissue remodeling activity likely involved during the transition from benign to early-stage melanoma. These processes include the gene expression of cancer associated fibroblasts (CAF), collagens, extracellular matrix (ECM) and integrins, which assist early melanoma development and the immune surveillance which plays a substantial role at this early stage. Further, we show genes upregulated in dysplastic nevi (DN) are also overexpressed in melanoma tissue, supporting the notion DN may serve as a transitional phase towards oncogenesis. Lastly, we reveal CN collected from healthy individuals exhibit different gene signatures compared to histologically benign nevi tissue located adjacent to melanoma (adjacent nevi, AN). We show the expression profile of micro-dissected AN tissue is more similar to melanoma compared to CN, revealing the melanoma influence on this annexed tissue.

Impact of the COVID-19 pandemic on melanoma diagnosis and presentation: a `review.

The ongoing challenges posed by COVID-19 are concerning for their impact on successful detection and recognition of melanoma as total body skin examinations and skin biopsies are critical for identifying early-stage melanoma and intervening before progression to metastatic disease. A comprehensive electronic search of PubMed/MEDLINE was conducted on or before August 1, 2022, using the search terms ("skin" AND "COVID-19"), (["skin cancer" AND "COVID-19"] OR ["skin cancer" AND "coronavirus"]), (["melanoma" AND "COVID-19"] OR ["melanoma" AND "coronavirus"]), ("dermatology" AND "COVID-19"), and ("cutaneous" AND "COVID-19"). Eight articles representing Belgium, Chile, France, Germany, Spain, the United Kingdom, and the United States were included. Four articles analyzed changes in the proportion of in situ melanoma at diagnosis and consistently reported decreases, with an overall decrease ranging from 7.6 to 40.4%. Five studies analyzed changes in the proportion of melanoma diagnoses by staging, but no clear changes in staging patterns were observed. Five studies analyzed changes in the mean Breslow thickness of melanoma diagnoses and consistently reported increases, with an overall increase ranging from 4.0 to 38%. Disruptions to proper diagnosis and treatment of melanoma are creating undue morbidity, mortality, and healthcare costs as the pandemic continues. Continued research with improved, centralized data collection is needed to better address the COVID-19 pandemic's ongoing challenge to appropriate detection and treatment of melanoma.

1235 Endothelial AMBRA1 and Claudin5 as potential biomarkers of metastases in early-stage melanoma

1235 Endothelial AMBRA1 and Claudin5 as potential biomarkers of metastases in early-stage melanoma

Current application of immunotherapy in melanoma.

Current application of immunotherapy in melanoma.

1232 Quantitative proteomics on laser microdissected early-stage melanomas reveals a mitochondrial involvement in the disease progression

1232 Quantitative proteomics on laser microdissected early-stage melanomas reveals a mitochondrial involvement in the disease progression

Melanoma Clinical Decision Support System: An Artificial Intelligence-Based Tool to Diagnose and Predict Disease Outcome in Early-Stage Melanoma Patients.

This study set out to assess the performance of an artificial intelligence (AI) algorithm based on clinical data and dermatoscopic imaging for the early diagnosis of melanoma, and its capacity to define the metastatic progression of melanoma through serological and histopathological biomarkers, enabling dermatologists to make more informed decisions about patient management. Integrated analysis of demographic data, images of the skin lesions, and serum and histopathological markers were analyzed in a group of 196 patients with melanoma. The interleukins (ILs) IL-4, IL-6, IL-10, and IL-17A as well as IFNγ (interferon), GM-CSF (granulocyte and macrophage colony-stimulating factor), TGFβ (transforming growth factor), and the protein DCD (dermcidin) were quantified in the serum of melanoma patients at the time of diagnosis, and the expression of the RKIP, PIRIN, BCL2, BCL3, MITF, and ANXA5 proteins was detected by immunohistochemistry (IHC) in melanoma biopsies. An AI algorithm was used to improve the early diagnosis of melanoma and to predict the risk of metastasis and of disease-free survival. Two models were obtained to predict metastasis (including "all patients" or only patients "at early stages of melanoma"), and a series of attributes were seen to predict the progression of metastasis: Breslow thickness, infiltrating BCL-2 expressing lymphocytes, and IL-4 and IL-6 serum levels. Importantly, a decrease in serum GM-CSF seems to be a marker of poor prognosis in patients with early-stage melanomas.

Abstract 6295: Belzupacap sarotalocan, an investigational virus-like drug conjugate, preferentially binds cancer cell glycosaminoglycans associated with the epithelial to mesenchymal transition

Abstract Human papillomaviruses (HPVs) are non-enveloped, double-stranded DNA viruses that utilize modified glycosaminogycans (GAGs) such as heparan sulfate proteoglycans, within the epidermal basement membrane as initial attachment factors prior to cell entry and infection. Similarly modified GAGs are also commonly found on the surface of tumor cells, enabling HPV capsids with tumor-selective binding properties. Belzupacap sarotalocan (bel-sar, formerly AU-011), is a virus-like drug conjugate composed of an HPV virus-like particle (VLP) conjugated to a cytotoxic payload (phthalocyanine dye) which upon light activation, causes rapid immunogenic tumor necrosis of bound tumor cells in vivo and has the potential to induce long-term anti-tumor immunity. Bel-sar has been shown to bind to and kill a wide variety of tumor types in preclinical models and is entering a Phase 3 clinical trial for the treatment of early-stage choroidal melanoma. To investigate the precise GAG modifications as well as the overall genetic signature which mediates the binding specificity of bel-sar, a large, multi-tumor type binding screen of bel-sar on 124 cancer cell lines was performed in vitro, where we identified a strong relationship between drug binding and pathways involved in the epithelial to mesenchymal transition (EMT). Additionally, through dose escalation treatments, HeLa subclones were generated which fail to bind and are resistant to the cytotoxic effects of bel-sar. Gene expression profiling and biochemical characterization of these resistant HeLa cell lines demonstrated a strong reduction in the expression of GAGs and a marked down-regulation of TGF-β signaling, ultimately resulting in the induction of a mesenchymal to epithelial transition (MET). Furthermore, gene-gene correlation studies showed that a variety of GAG biosynthesis genes are associated with TGF-β signaling and EMT progression, and that these genes are important determinates of bel-sar binding. Overall, these data suggest that the binding of bel-sar and HPV VLPs is strongly dependent on the expression and modification of GAGs that occurs during the EMT process. As many cancers, especially metastatic tumors, are thought to undergo at least partial EMT, these data provide mechanistic insights into the tumor-targeting ability of HPV VLP-conjugates such as bel-sar and suggest possible applications for such therapies across a wide variety of other tumor types. Citation Format: Nathan R. Fons, Rhonda C. Kines, John T. Schiller. Belzupacap sarotalocan, an investigational virus-like drug conjugate, preferentially binds cancer cell glycosaminoglycans associated with the epithelial to mesenchymal transition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6295.

Dermatosurgery - from surgical option to integral part of dermatologic therapy.

Dermatosurgery was long considered an isolated, and not always important, discipline within dermatology. As a therapeutic option, it was considered either the gold standard of first-line therapy, for example in basal cell carcinoma surgery and treatment of early-stage melanoma, or the last option, for instance in the treatment of warts. The fact that a profound change has taken place and that dermatosurgery is now an integral, equal, sometimes leading and always significant component of dermatology will be demonstrated in this review using three examples from geriatric dermatology, the treatment of hidradenitis suppurativa (acne inversa), and melanoma therapy. This review is supplemented by a section on the most important technique in dermatosurgery: microscopic (micrographic) surgery or Mohs Surgery.

Factors Influencing General Dermatologists When Referring Patients With Head and Neck Melanoma for Mohs Micrographic Surgery: A Nationwide Cross-Sectional Survey.

Mohs micrographic surgery (MMS) for cutaneous melanoma has demonstrated higher cure rates, lower local recurrence rates, and improved survival compared with wide local excision (WLE). However, factors affecting referrals by general dermatologists for MMS of head and neck melanoma (HNM) are unknown. To elucidate referral factors and treatment perspectives of general dermatologists regarding MMS for melanoma in situ (MIS)/lentigo maligna (LM) and early-stage melanoma on the head and neck. A cross-sectional analysis was performed using survey responses of general dermatologists with membership in the American Academy of Dermatology . A total of 231 and 132 of the 402 responding general dermatologists routinely referred melanoma in situ MIS/LM and early invasive melanoma for MMS, respectively. Lack of local access to a Mohs surgeon was the most common deterring reason for MIS/LM referral to MMS, whereas the preference for WLE was the most common deterring reason for early invasive melanoma. Lack of local access to a Mohs surgeon treating HNM with MMS is the primary barrier in referrals to Mohs surgeons for MIS and LM. Among general dermatologists, WLE is preferred for early invasive HNM.

Conventional versus Reduced-Frequency Follow-Up in Early-Stage Melanoma Survivors: A Systematic Review with Meta-Analysis

To date, there have been multiple studies and clinical guidelines or recommendations for complex management of melanoma patients. The most controversial subjects included the frequency of follow-up. This study provides a coherent and comprehensive comparison of conventional vs. reduced-frequency follow-up strategies for early-stage melanoma patients. The value of our study consists in the precise analysis of a large collection of articles and the selection of the most valuable works in relation to the topic according to rigorous criteria, which allowed for a thorough study of the topic. The search strategy was implemented using multiple databases. The inclusion criteria were randomized clinical trial or cohort studies that compared the outcomes of a conventional follow-up schedule versus a reduced-frequency follow-up schedule for patients diagnosed with melanoma. In this study, authors analyzed recurrence and 3-year survival. Meta-analysis of outcomes presented by Deckers et al. and Moncrieff et. al. did not reveal a significant difference favoring one of the groups (OR 1.14; 95%CI: 0.65-2.00; p = 0.64). The meta-analysis of 3-year overall survival included two studies. The statistical analysis showed no significant difference in favor of the conventional follow-up group. (OR 1.10; 95%CI: 0.57-2.11; p = 0.79). Our meta-analysis shows that there is no advantage in a conventional follow-up regimen over a reduced-frequency regimen in early-stage melanoma patients.

Dual Role of DUOX1-Derived Reactive Oxygen Species in Melanoma

Melanoma is the most serious type of skin cancer. Inflammation and oxidative stress play an essential role in the development of several types of cancer, including melanoma. Although oxidative stress promotes tumor growth, once cells escape from the primary tumor, they are subjected to a more hostile environment, with higher levels of oxidative stress typically killing most cancer cells. As Dual Oxidase 1 (DUOX1) is a major producer of reactive oxygen species (ROS) in epithelia, we used allotransplantation and autochthonous melanoma models in zebrafish together with in silico analysis of the occurrence and relevance of DUOX1 expression of the skin cutaneous melanoma (SKCM) cohort of The Cancer Genome Atlas (TCGA) to address the role of this enzyme in the aggressiveness of melanoma cells in vivo. It was found that high transcript levels of the gene encoding DUOX1 were associated with the poor prognosis of patients in the early-stage melanoma of TCGA cohort. However, DUOX1 transcript levels were not found to be associated to the prognosis of late-stage SKCM patients. In addition, the transcript level of DUOX1 in metastatic SKCM was lower than in primary SKCM. Using zebrafish primary melanoma and allotransplantation models, we interrogated the role of DUOX1 in vivo. Our results confirmed a dual role of DUOX1, which restrains melanoma proliferation but promotes metastasis. As this effect is only observed in immunocompromised individuals, the immune system appears to be able to counteract this elevated metastatic potential of DUOX1-deficient melanomas.

Preoperative Plasma miRNA Levels Predict Prognosis in Early-stage Malignant Melanoma.

Non-invasive circulating tumor biomarkers in liquid biopsy, such as microRNAs (miRNA), provide for better personalization of treatment strategies. The aim of our study was to assess the prognosis of patients with melanoma undergoing tumor resection with curative intent based on analysis of selected circulating miRNAs. A total of 22 patients with stage I to III melanoma were enrolled into this prospective study. Plasma samples were obtained pre-surgery and early post-surgery from peripheral blood draws. A panel of 23 candidate miRNAs was designed and expression of miRNAs were analyzed by reverse transcription-quantitative polymerase chain reaction with exogenous reference control cel-miR-39-3p. Higher preoperative expression levels of miR-99a (p=0.008), miR-320 (p=0.009), miR-1908 (p=0.001), miR-494 (p=0.018) and miR-4487 (p=0.048) were associated with a shorter disease-free interval. Similarly, higher preoperative plasma levels of miR-99a (p=0.017), miR-221 (p=0.026), miR-320 (p=0.016), miR-494 (p=0.009), miR-1260 (p=0.026) and miR-1908 (p=0.024) were associated with worse overall survival. No significant differences between pre- and postoperative plasma miRNA levels were observed. Liquid biopsy is a minimally-invasive approach which can lead to a better understanding of cancer behavior and offers the possibility of precise patient prognosis, allowing selection of the most appropriate treatment. Our study showed that preoperative plasma levels of miR-99a, miR-221, miR-320, miR-494, miR-1908 and miR-4487 were associated with disease-free interval and overall survival of patients with early-stage melanoma. This approach may help in decision-making about the appropriateness of modern adjuvant treatment administration in patients with resectable melanoma.

A genome-wide association study of germline variation and melanoma prognosis.

The high mortality of cutaneous melanoma (CM) is partly due to unpredictable patterns of disease progression in patients with early-stage lesions. The reliable prediction of advanced disease risk from early-stage CM, is an urgent clinical need, especially given the recent expansion of immune checkpoint inhibitor therapy to the adjuvant setting. In our study, we comprehensively investigated the role of germline variants as CM prognostic markers. We performed a genome-wide association analysis in two independent cohorts of N=551 (discovery), and N=550 (validation) early-stage immunotherapy-naïve melanoma patients. A multivariable Cox proportional hazard regression model was used to identify associations with overall survival in the discovery group, followed by a validation analysis. Transcriptomic profiling and survival analysis were used to elucidate the biological relevance of candidate genes associated with CM progression. We found two independent associations of germline variants with melanoma prognosis. The alternate alleles of these two SNPs were both associated with an increased risk of death [rs60970102 in MELK: HR=3.14 (2.05-4.81), p=1.48×10-7; and rs77480547 in SH3BP4: HR=3.02 (2.02-4.52), p=7.58×10-8, both in the pooled cohort]. The addition of the combined risk alleles (CRA) of the identified variants into the prognostic model improved the predictive power, as opposed to a model of clinical covariates alone. Our study provides suggestive evidence of novel melanoma germline prognostic markers, implicating two candidate genes: an oncogene MELK and a tumor suppressor SH3BP4, both previously suggested to affect CM progression. Pending further validation, these findings suggest that the genetic factors may improve the prognostic stratification of high-risk early-stage CM patients, and propose putative biological insights for potential therapeutic investigation of these targets to prevent aggressive outcome from early-stage melanoma.

Abstract IA024: Cellular plasticity during metastatic colony formation in patients

Abstract Systemic cancer spread is a process whose complexity is only poorly understood. We have learned over the past years that metastatic dissemination starts very early in the evolution of a malignant clone of most cancers and that disseminated cancer cells (DCC) continue to acquire genomic alterations at the distant site. Consequently, it seems plausible that cellular phenotypes change during metastatic colony formation as well, although the impact of genotype changes on the cellular phenotypes is not clear at all. We studied this process in malignant melanoma, exploiting the unique chance that routine sentinel lymph node (SLN) biopsies provide for a cancer that is diagnosed much earlier and at smaller size than most other cancers. Moreover, mouse models suggest that lymph nodes serve as exit routes for systemic dissemination of cancer cells and that their colonization indues tumor-immune tolerance promoting distal metastasis. Melanoma DCC enter SLNs and may or may not give rise to a metastatic colony. We therefore first assessed in a large patient cohort comprising more than 500 early-stage melanoma patients, which melanoma marker is best suited to identify candidate metastasis founder cells and then progressed to provide their detailed molecular characterization. We found that melanoma cellular phenotypes indeed change over metastatic colony formation, indicating that differential molecular stage-specific targeting may be required to prevent metastasis formation. Of note, we observed mechanisms of very early immune suppression via DCC-derived EVs suggesting early DCC must find immune-escape strategies at the metastatic site. Citation Format: Melanie Werner-Klein. Cellular plasticity during metastatic colony formation in patients [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr IA024.