Psoriasis and eczema are chronic inflammatory skin diseases with overlapping histopathological features, which often lead to diagnostic uncertainty even among experienced dermatopathologists. To address this challenge, we developed a computer-assisted diagnostic framework that combines the Virchow foundation model, pretrained on 1.5 million whole-slide images, with multi-instance learning (MIL) to classify psoriasis and eczema from digitized histopathology slides. Using an internal dataset (n = 40) and an external validation cohort (n = 40), equally balanced between both conditions and annotated by board-certified dermatopathologists, our best-performing configuration (Virchow + CLAM) achieved 85% accuracy, a macro-averaged F1 score of 0.80, and an AUC of 0.81 on the external cohort. This substantially outperformed baseline convolutional neural networks, which reached 61% accuracy, and models relying solely on pretrained feature extractors without MIL, which achieved an average accuracy of 68.8%. In a reader study on the same external cohort, individual dermatopathologist accuracies ranged from 47.5 to 70.0%, with a majority-vote consensus accuracy of 62.5%; our method outperformed both the average individual reader and the consensus under histology-only conditions. Furthermore, the model generates attention heatmaps that provide supportive visual context by highlighting regions associated with model predictions. Importantly, this study is designed as a methodological proof-of-concept conducted under controlled, histology-only conditions and is not intended for direct clinical deployment. Rather than demonstrating clinical readiness, it illustrates the potential of domain-specific foundation models combined with MIL for addressing diagnostically challenging inflammatory dermatoses.

DNA methylation provides a stable record of cellular identity, capturing epigenetic programs that distinguish specialized cell states despite a shared genome. Because malignant transformation and tumour progression are accompanied by extensive epigenetic remodeling, we hypothesized that the methylome of melanocytic lesions contains biologically and clinically relevant information for both diagnosis and disease progression. In a cohort of 1,001 tissue samples prospectively collected across eight German university hospitals profiled using Illumina Infinium MethylationEPIC arrays, we compared machine-learning models based on selected Cytosine phosphate Guanine (CpG) methylation sites with models incorporating biology-guided features, including epigenetic age acceleration, cell type composition and copy-number variation burden. In an external test set, the best diagnostic classifier was CpG-based and distinguished melanocytic nevi, noninvasive melanoma and invasive melanoma with a macro-averaged area under the receiver operating characteristic curve of 0.919 (95

BACKGROUND:Patients with BRAF wildtype metastatic melanoma who exhibit primary resistance to immune checkpoint inhibitors (ICIs) have a poor prognosis. Chemotherapy has been shown to induce genetic mutations, modify the tumour microenvironment and microbiome, and influence immune system activity. OBJECTIVES:To investigate in a prospective multicentre phase II trial whether two applications of an alkylating agent (dacarbazine) can sensitize patients with metastatic melanoma who are nonresponsive to ICIs to the same checkpoint inhibitor regime. METHODS:The PROMIT trial (NCT04225390) enrolled patients with histologically confirmed BRAF wildtype metastatic melanoma who exhibited primary resistance to ICI therapy. Following radiological evidence of primary resistance to ICI (ipilimumab + nivolumab or pembrolizumab) upon the first staging after initiation, patients received two doses of dacarbazine at 850 mg m-2 intravenously on days 1 and 21. Subsequently, 1 week after application of the second dose of dacarbazine, patients were rechallenged with the same ICI therapy to which they had previously shown progressive disease. RESULTS:In total, 53 patients were enrolled across four German skin cancer centres. Of these patients, 38 were evaluable for efficacy, having received at least one dose of ICI re-exposure. The overall objective response rate was 18% (95% confidence interval 0.08-0.34), with 7 of 38 patients achieving a partial response. The disease control rate was 37%. Therapy was well tolerated, with treatment-related ≥ grade 3 CTCAE adverse events occurring in 10% of patients. No new safety signals were observed. CONCLUSIONS:The study indicates that short-term chemotherapy followed by ICI rechallenge can overcome primary ICI resistance in patients with melanoma, supporting its potential as a new therapeutic option in clinical practice.

Immune checkpoint inhibitors (ICIs) have substantially improved outcomes in advanced melanoma but are frequently linked to immune-related adverse events (irAEs). Vitiligo is a common cutaneous irAE and has been consistently associated with improved patient outcome, including prolonged progression-free and overall survival. It also represents significant visual stigma, particularly when the face is involved. Traditional treatment comprises topical steroids, calcineurin inhibitors, laser, and phototherapy which often have insufficient effects. Since 2023, the first approved drug for non-segmental vitiligo (NSV) with facial involvement, the topical Janus kinase inhibitor ruxolitinib, has been available. However, experience with its use in ICI-induced vitiligo remains limited. In this exploratory analysis, three patients who developed facial vitiligo following ICI therapy applied 1.5% ruxolitinib cream to affected facial areas twice daily. After six (two patients), and twelve months (one patient), extensive repigmentation was observed, quantified at 95.7%, 78.9%, and 99.1% using a novel semi-automatic tool. Quality-of-life questionnaires showed mean reductions of 57.6% (Vitiligo DLQI) and 68.2% (Vitiligo-specific Quality of Life) in disease burden. Treatment was associated with substantial repigmentation without observed side effects. Further evaluation in larger, prospective cohorts is warranted to better define treatment effects, clinical applicability, and long-term safety.

The current classification of sweat gland carcinomas is based on histomorphological characteristics and distinguishes between more than 20 entities. Most patients are older, but some subtypes also affect middle-aged and younger patients. The majority of tumors arise de novo. Sweat gland carcinomas have nonspecific clinical features. The tumors are usually located in the head and neck area and on extremities, with the exception of extramammary Paget's carcinoma, which has an anogenital predilection. Sweat gland carcinomas occurring in the armpit may pose a histomorphological challenge in distinguishing them from metastatic or primary breast cancer. The diagnosis is made histopathologically via excisional biopsy. Histopathological subdifferentiation is essential for an accurate diagnosis. Metastasis initially occurs locally (per continuitatem), later to regional lymph nodes and distant organs. The treatment of choice is complete surgical excision with entire histopathological margin control (microscopically controlled surgery) or alternatively with wide local excision with a safety margin. Postoperative adjuvant radiotherapy is recommended for high-risk tumors. The data available for drug therapy of advanced tumors is generally weak, with the best results seen for chemotherapy using platinum derivatives. Furthermore, targeted therapies are possible, e.g., with HER2/neu or EGFR inhibitors, possibly in combination with chemotherapy. A risk-adapted regimen is recommended for the follow-up care.

BACKGROUND AND OBJECTIVES:Complete lymph node dissection (CLND) is the standard of care in patients with regional nodal melanoma macrometastasis. However, evidence on surgical procedures in the era of adjuvant systemic therapies is lacking. PATIENTS AND METHODS:This retrospective multi-center study included stage IIIB-D melanoma patients with nodal macrometastasis undergoing CLND or selective lymph node extirpation (LNE) prior to adjuvant therapy. CLND and LNE were compared regarding recurrence-free survival (RFS), nodal metastasis-free survival (NFS) and overall survival (OS). RESULTS:320 melanoma patients were included (median age 62; 55.5% male). Patients received PD-1 monotherapy (77.8%), targeted therapy (21.3%) or both sequentially (0.9%), as well as adjuvant radiotherapy in 40.9%. RFS and OS did not differ significantly between patients receiving CLND or LNE, while NFS was significantly prolonged following CLND (HR 0.3917; p = 0.005). After adjustment for risk factors by multivariate Cox regression, a prolonged RFS for CLND vs. LNE was found (HR 0.676; p = 0.04), but no benefit for OS. The rate of complications was significantly higher in the CLND group. CONCLUSIONS:CLND showed no OS benefit compared to LNE, while local control was improved. CLND can be recommended in the context of adjuvant therapies, however, the increased rate of complications should be considered.

BACKGROUND:Patients with advanced melanoma progressing after immune checkpoint inhibition (ICI) and BRAF/MEK inhibition have limited therapeutic options. In the LEAP-004 trial, pembrolizumab plus lenvatinib demonstrated activity in PD-1-refractory melanoma. The combination has emerged as a potential option when approved therapies have been exhausted; however, real-world evidence regarding its efficacy remains limited. METHODS:This retrospective, multicenter DeCOG study included patients with advanced melanoma treated with anti-PD-1 plus lenvatinib after failure of anti-PD-1-based therapy at 11 major skin-cancer centers in Germany and Switzerland between October 2020 and March 2025. RESULTS:Overall, 120 patients were analyzed (median age 59 years); 70 % were male and 38 % had an ECOG performance status > 1. Most patients had ≥ 3 metastatic sites (69 %), brain metastases (42 %), and elevated LDH (58 %). Median follow-up was 13.4 months. Patients received a median of two prior systemic therapy lines; 98 % had been pretreated with ipilimumab/nivolumab, and 66 % exhibited primary resistance to prior ICI. The objective response rate was 23 %, with a median duration of response of 10 months; disease control rate was 47 %. Median progression-free survival (mPFS) was 4 months and median overall survival (mOS) was 10 months, with 12-month PFS and OS rates of 17 % and 42 %, respectively. Durable disease control beyond six months was observed in 23 % of patients, with mPFS of 21 months. Grade ≥ 3 treatment-related adverse events occurred in 21 % of patients. CONCLUSIONS:In this real-world cohort, anti-PD-1 plus lenvatinib demonstrated meaningful efficacy in a subset of heavily pretreated patients, predominantly in those with BRAF wild-type melanoma.

BACKGROUND:Distinguishing hand eczema from palmar psoriasis is a common diagnostic challenge due to overlapping clinical and histopathological features. OBJECTIVE:This study aimed to validate morphological and immunohistochemical criteria for differentiating these two conditions using the digital pathology tools QuPath and ImageJ. METHODS:One hundred forty-two histological samples with confirmed clinical diagnoses were stained with hematoxylin and eosin and subjected to immunohistochemical staining for CD3, CD15, CD20, CD123, S100, and PHH3. The samples were digitized for analysis. QuPath was used to automate annotation, segment epidermal layers, and measure rete ridge elongation, width, and suprapapillary epidermal thickness. Immunohistochemical markers were also analyzed using QuPath. ImageJ was employed to quantify spongiosis using color thresholding techniques. RESULTS:Suprapapillary epidermal thickness was significantly lower in psoriasis compared to eczema (p < 0.001; AUC = 0.72). Rete ridge elongation (p = 0.002; AUC = 0.704) and width (p < 0.001; AUC = 0.698) also showed significant differences. Furthermore, hypogranulosis was more pronounced in psoriasis (p = 0.012; AUC = 0.602), while S100-positive cells in the epidermis were more commonly observed in eczema (p = 0.013; AUC = 0.583). CONCLUSION:Quantitative assessment of suprapapillary epidermal thickness and rete ridge morphology offers a reliable and objective method for differentiating palmar psoriasis from hand eczema, enhancing diagnostic accuracy.

PURPOSE:Adjuvant treatment with immune checkpoint inhibition (PD-1) and targeted therapy (TT) with BRAF + MEK inhibitors significantly improved recurrence-free survival (RFS) of patients with stage III melanoma. We investigated efficacy of adjuvant therapy with PD-1 or TT under real-world conditions. MATERIALS AND METHODS:A total of 589 patients with stage III melanoma who started adjuvant PD-1 or TT between June 2018 and September 2019 from 11 major German Dermatologic Cooperative Oncology Group skin cancer centers were followed for 4 years. End points were RFS, overall survival (OS), and melanoma-specific survival. Survival analyses and adjusted hazard ratios (HRs) were estimated with Kaplan-Meier and Cox proportional hazards model, inverse probability treatment weighting, and propensity score matching. RESULTS:RFS at 48 months was 42.9% (95% CI, 38.5 to 47.8) for all PD-1 patients and 52.6% (95% CI, 43.6 to 63.3) for TT patients. Among patients with BRAF mutation, rate of recurrence was higher for PD-1 compared with TT (HR, 1.57 [95% CI, 1.09 to 2.26]). OS at 4 years was 80.8% (95% CI, 73.6 to 88.7) for PD-1-treated patients with BRAF mutation and 87.3% (95% CI, 81.0 to 94.0) for TT patients. Patients starting adjuvant PD-1 after resection of macroscopic lymph node metastases had a higher risk of rapid recurrence (1-year RFS all PD-1 58%) compared with 87% in TT patients. Rate of recurrence after premature discontinuation (≤6 v >6 months treatment) was higher in TT patients (HR, 1.47 [95% CI, 0.67 to 3.23]), but not in PD-1 patients (HR, 1.07 [95% CI, 0.73 to 1.55]). CONCLUSION:PD-1-treated patients with BRAF mutation had a markedly higher rate of relapse compared with TT patients. Rapid recurrences occurred particularly in PD-1-treated patients with previous macroscopic lymph node metastasis. Treatment duration shorter than 6 months did not negatively affect RFS in PD-1, but in TT patients.

Background CV8102, a toll-like receptor 7/8 and RIG I agonist, has demonstrated antitumor immune responses in preclinical studies. We investigated intratumoral (IT) administration of CV8102 in patients with anti-programmed cell death protein-1 (PD-1) therapy-naïve or anti-PD-1 therapy-refractory cutaneous melanoma (cMEL) and in patients with advanced cutaneous squamous cell carcinoma, head and neck squamous cell carcinoma and adenoid cystic carcinoma.Methods This open-label, cohort-based, phase I dose escalation study aimed to establish the maximum tolerated dose (MTD), recommended dose (RD), safety and preliminary efficacy of CV8102 as monotherapy or in combination with a PD-1 inhibitor. The preliminary efficacy of the RD was assessed in patients with cMEL in the expansion cohorts.Results Between September 2017 and October 2022, 98 patients were enrolled in monotherapy and combination therapy dose escalation and dose expansion cohorts. Two patients in the CV8102 monotherapy dose escalation cohort experienced relevant toxicities at the 900 µg dose level. One patient had Grade 3 aspartate transaminase/alanine aminotransferase elevation which met dose-limiting toxicity (DLT) criteria. Another patient experienced Grade 3 immune-mediated pneumonitis. No DLTs occurred in the combination therapy dose escalation cohort. The MTD was not formally reached and the RD for expansion was 600 µg. Common treatment-emergent adverse events were fever (57%), chills (37%) and fatigue (25%). In the dose escalation part, objective responses occurred in 3/33 patients treated with CV8102 as monotherapy and in 2/25 patients treated with CV8102 plus a PD-1 inhibitor. In the expansion cohorts in patients with anti-PD-1 therapy-refractory melanoma, 0/10 patients treated with CV8102 as monotherapy and 5/30 patients (17%) treated in combination with a PD-1 inhibitor experienced objective responses.Conclusions IT CV8102 was generally well tolerated with preliminary signs of efficacy as monotherapy and in combination with a PD-1 inhibitor.Trial registration number NCT03291002.

Chondroitin sulfate proteoglycan 4 (CSPG4) is a promising target for melanoma immunotherapy, but its expression in benign melanocytic lesions and its diagnostic value remain unexplored. This study assessed CSPG4 expression in benign nevi (BN), dysplastic nevi (DN), and superficial spreading melanomas (SSM), comparing it with PRAME (PReferentially expressed Antigen in MElanoma) and evaluating the cell division cycle 7-related protein kinase (CDC7) and the proliferation marker Ki67. Histological sections were stained using automated instruments, digitized, and analyzed using QuPath. Cohorts of BN, DN, and SSM were created, and positive cells/mm2 and H-scores were determined. A total of 336 IHC stainings from 84 specimens were analyzed. CSPG4 expression was readily detected in SSM and was significantly stronger in DN (p = 0.005), with the highest intensity observed in BN (p < 0.001). PRAME showed the highest density of positive cells/mm2 in SSM, was significantly reduced in DN (p < 0.001), and was lowest in BN (p < 0.001). Ki67 expression was strong in SSM, moderate in DN (p = 0.62), and low in BN (p = 0.008). CDC7 expression was most intense in DN, less in SSM (p = 0.39), and weakest in BN (p = 0.002). ROC AUC values for SSM versus DN and SSM versus BN were 0.764 and 0.921 for CSPG4, 0.85 and 0.889 for PRAME, 0.735 and 0.742 for Ki67, and 0.425 and 0.767 for CDC7. While PRAME was the most reliable marker for differentiating DN and SSM, CSPG4 was superior for distinguishing BN from SSM due to its high expression in BN. However, CSPG4-targeting therapies may trigger on-target/off-tumor effects due to its high expression in melanocytic nevi. © 2025 The Author(s). The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Hintergrund Dermatologische orale Antitumortherapeutika (OAT) sind h & auml;ufig interaktionsanf & auml;llig und werden in komplexen Therapieschemata eingesetzt. Das klinisch-pharmakologische/pharmazeutische Therapiebegleitungskonzept der randomisierten AMBORA-Studie verbesserte die Arzneimitteltherapiesicherheit bei verschiedensten OAT signifikant, dermato-onkologische Patienten waren jedoch nicht eingeschlossen. Das Konzept wurde im Anschluss in die klinische Routine implementiert und die Dermato-Onkologie eingeschlossen. Ziel dieser Untersuchung war die Analyse von Medikationsfehlern sowie der Adh & auml;renz bei Patienten mit dermatologischen OAT.Patienten und Methodik Medikationsfehler wurden unter anderem nach Ursache charakterisiert (PCNE V9.1). Die Adh & auml;renz wurde mit dem ,,Medication-Event-Monitoring-System" MEMS (R) Button und dem MARS-D Fragebogen erfasst. Prim & auml;re Endpunkte waren der Anteil an gel & ouml;sten OAT-bezogenen Medikationsfehlern und die Dosing Adherence (DA), (Tage mit korrekter OAT-Einnahme) & uuml;ber 12 Wochen.Ergebnisse Bei 92 Patienten (81,5% mit Melanom) wurden im Mittel 1,6 Medikationsfehler pro Patient festgestellt. Davon betrafen 61,6% die OAT, von denen 89,2% behoben wurden. Von 52 Patienten, die am zus & auml;tzlichen Adh & auml;renzmonitoring teilnahmen, waren 48 evaluierbar und erreichten eine mediane DA von 95,0% sowie einen MARS-D-Score von 25/25. Die DA war bei OAT mit 1 x t & auml;glicher Einnahme h & ouml;her als bei 2 x t & auml;glicher (p = 0,0127).Schlussfolgerungen Das interprofessionelle AMBORA-Therapiebegleitungskonzept in der Dermato-Onkologie war mit einem gro ss en Anteil gel & ouml;ster Medikationsfehler und hoher Adh & auml;renz assoziiert. Evidenzbasiertes Medikationsmanagement und Patientenberatungen und Patientenberatung durch klinische Pharmakologen/Pharmazeuten optimieren die Arzneimitteltherapiesicherheit in der dermato-onkologischen Versorgung.

Dear Editor, In recent years, treatment with immune checkpoint inhibitors (ICI) has revolutionized cancer therapy. Monoclonal antibodies that block immune checkpoint receptors such as cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1) or its ligand PD-L1 prevent the tumor from suppressing adaptive immune responses.1 Especially PD-L1 is expressed on tumor cells, but also on various healthy cell types.2 When we evaluated a combination therapy of active immunization via dendritic cells (DCs) and ICIs in the context of Merkel cell carcinoma (MCC) in order to improve the efficacy of the treatment, we observed an unexpected inhibitory effect of the anti-PD-L1 ICI Avelumab. In cell culture experiments with the most commonly used type of DCs in clinical trials on therapeutic cancer vaccination,3 the priming capacity of the tumor antigen-loaded human monocyte-derived DCs was reduced in the presence of Avelumab but not Pembrolizumab (anti-PD-1; Figure S1A). Experiments with GFP-expressing DCs showed a disappearance of the DCs in co-cultures with autologous lymphocytes and Avelumab over time (data not shown). However, no direct toxic effect of the antibody against pure DCs was observed (Figure S1B). Moreover, the DCs' ability to stimulate T cell receptor-transfected pure CD8+ T cells was not influenced by the ICI antibodies (Figure S1C). From these unanticipated findings, we concluded that Avelumab, in the presence of autologous lymphocytes, had a detrimental effect on the DCs. In contrast to other approved anti-PD-L1 antibodies like Atezolizumab and Durvalumab, Avelumab contains a constant region (Fc-part) of the IgG1 isotype that is capable of inducing antibody-dependent cellular cytotoxicity (ADCC) against PD-L1 expressing tumor cells, which has been shown to be beneficial in preclinical studies.4 However, healthy cells including DCs also express PD-L1. Therefore, Avelumab could induce an unwanted ADCC reaction against the DCs, as any antibody with a suitable Fc-part that binds efficiently to these cells would do. Flow cytometry experiments showed that monocyte-derived DCs expressed high levels of PD-L1 and that Avelumab, Atezolizumab, and Durvalumab but not Pembrolizumab efficiently bound to the DCs (see Figure 1A). Also, the human primary DC subpopulations cDC1, DC2, and DC3, which are responsible for antigen presentation,5 expressed PD-L1 upon Toll-like receptor 7/8 stimulation with R848 (see Figure 1A). Primarily, two types of cells exert ADCC: macrophages and natural killer (NK) cells. Since the latter are present in substantial numbers in the lymphocyte fractions we used, we examined whether this was the specific cell type that carried out an Avelumab-dependent ADCC against DCs. In a classical chromium-release cytotoxicity assay, we were able to show that in the presence of Avelumab, DCs were efficiently lysed by purified NK cells whereas intermediate lysis was seen when complete peripheral blood mononuclear cells (PBMCs) were used as effector cells (Figure 1B). Consequently, NK-depleted PBMCs showed no lysis. Likewise, there was also no cytotoxicity observed in the control conditions containing no antibody or the anti-PD-1 antibody Pembrolizumab (Figure 1B). The Fc-part-mutated antibodies Durvalumab and Atezolizumab were also not able to induce lysis of the DCs (Figure 1C), although they bound to the DCs to a similar extent (Figure 1A). This clearly shows that Avelumab induced an efficient ADCC reaction by NK cells against autologous DCs. In the experiments described above, monocyte-derived cytokine-matured DCs were used, as these cells are the most commonly used in clinical trials for DC-based therapeutic tumor vaccination. Hence, we examined whether the maturation type and state influenced the susceptibility of DCs to Avelumab-mediated ADCC. We observed that DCs matured with several different stimuli (cytokine cocktail, lipopolysaccharide (LPS), R848, or polyinosinic:polycytidylic acid [poly I:C]) all expressed high levels of PD-L1. Even immature monocyte-derived DCs expressed PD-L1, however to a lower extent. Therefore, the cytotoxicity assay was repeated with purified NK cells and immature DCs, as well as DCs treated with the various maturation stimuli. In the presence of Avelumab all differently matured DCs and even immature DCs were lysed with similarly high efficiency (Figure 1D). This implies that the observed phenomenon may be of general relevance as it seems to apply to all cells expressing a sufficient level of PD-L1. Next to classical ADCC, additional antibody-mediated cytotoxic effects may exist in vivo, like complement-dependent cytotoxicity and antibody-dependent phagocytosis, which may aggravate the effect. In summary, we demonstrated that Avelumab-mediated ADCC via NK-cell activation can lead to the killing of DCs. These findings are of high clinical relevance for combination therapies with Avelumab. Therefore we recommend that patients receiving active immunotherapy like therapeutic vaccination should not simultaneously receive Avelumab treatment because this could result in an inhibitory effect on healthy PD-L1-expressing immune cells—especially when using ex vivo-generated DCs as vaccine. Either a sequential approach with initial vaccination and subsequent Avelumab treatment, or the use of Atezolizumab, Durvalumab, or Pembrolizumab instead of Avelumab is recommended. Avelumab is currently approved for MCC, urothelial carcinoma, and renal cell carcinoma. According to clinicaltrials.gov, Avelumab has been tested in hundreds of clinical trials from phase 1 to phase 3. Most of these combined the antibody with other treatment regimens such as chemotherapy, small molecule inhibitors, other therapeutic antibodies, oncolytic viruses, adoptive cell transfer, and therapeutic vaccination. Two trials used DCs together with Avelumab (NCT03707808 and NCT03152565) and both applied the DCs and the antibody simultaneously. Unfortunately, both did not address the question of whether the antibody had any effect on the DCs. Other trials used other types of vaccines, including peptides, adenoviral vectors, yeast formulations, and again, Avelumab was given at the same time as the vaccine. In a series of discontinued trials (QUILT-3 series) Avelumab was given together with allogenic NK-cells and a recombinant IL-15 superagonist. In one trial with highly progressed MCC patients (NCT03853317), this led to serious adverse events in more than half of the patients, and in another similar trial with pancreatic cancer patients (NCT03136406), one patient reported lymph node pain as an adverse event. In a breast cancer trial (NCT04215146) with paclitaxel and an oncolytic reovirus, the additional application of Avelumab resulted in fewer clinical responses but increased serious adverse events. However, in none of these trials, the effects of the antibody on DCs were examined. Hence, we think that awareness that Avelumab can probably kill important antigen-presenting cells will enable researchers to design more efficient treatment protocols for combination therapies with Avelumab. T.S., F.B., A.C.B., and L.H. performed experiments; T.S., F.B., and J.D. wrote the manuscript; N.C.B. and M.E. provided essential materials; T.S., J.D., N.S., and D.D. supervised experiments; A.C.B., L.H., D.D., N.B., M.E., C.B., and N.S. corrected the manuscript. All authors have read and approved the final manuscript. We appreciate Annett Hamann for helpful technical assistance. We thank the medical staff of the Uniklinikum Erlangen from the Dermatology Department especially Dr. Elias Koch for their support with all blood donations. Furthermore, we would like to thank all voluntary blood donors. We are grateful to Christian Ostalecki for his help in the production of the GFP-labelled Avelumab. We appreciate the working group of Thomas Harrer and especially Katja Schmidt, who supported us with the ELISPOT readouts. The authors declare no conflict of interest. This work was supported by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) via the Research Training Group GRK2504/1 (project number 401821119), research project B2 to Diana Dudziak and B4 to Jan Dörrie. The blood of healthy donors was obtained following informed consent and approval of the institutional review board (Ethics Committee of the Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany: Ref. no. 4158). The data generated in this study are available upon request from the corresponding author. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

RNA interference (RNAi) therapeutics represent breakthrough discoveries, but their use in cancer remains limited due to hepatocyte-specific targeting. Cancer metastasis is regulated by complex crosstalk between tumor cells and niche-derived factors. However, the molecular mechanisms enabling metastatic seeding and outgrowth in the liver remain incompletely understood, representing a major clinical challenge. We identified neuropeptide Y (NPY) as a promotor of liver metastasis. Hepatocyte-derived NPY attracts metastatic tumor cells to the liver niche. Subsequent microenvironment activation induces TGFβ, promoting a vicious cycle of perimetastatic NPY secretion and liver metastasis. Concomitantly, cancer cells upregulate the NPY-5 receptor (Y5R) which is correlated with liver metastasis. NPY-Y5R crosstalk drives chemotactic migration via cAMP and ERK signaling. Moreover, NPY-Y5R activation dephosphorylates checkpoint kinase 2 to promote clonogenicity and proliferation of cancer cells. Lipid nanoparticles (LNPs) are a promising drug delivery vehicle for siRNAs. LNPs carrying siRNA pools targeting NPY were designed, and preclinical studies provided evidence for efficacy for the treatment of liver metastasis. Our findings transform the limitation of hepatocyte specificity of RNA interference into a therapeutic advantage, introducing a paradigm for the treatment of hepatic metastases.

BACKGROUND:Immune checkpoint inhibitors (ICIs) have revolutionized melanoma treatment, with programmed cell death protein 1 (PD-1) inhibitors-alone or in combination with cytotoxic T-lymphocyte-associated protein 4 or lymphocyte-activation gene 3 inhibitors-demonstrating significant efficacy. However, there is a critical lack of robust data to determine the optimal sequencing of these therapies for individual patients. In particular, the role of relatlimab+nivolumab (rela/nivo) within treatment sequences remains poorly defined. Choosing the right sequence is strategic, as an inappropriate order may compromise the effectiveness of subsequent treatments and limit long-term benefits. METHODS:This multicenter retrospective and prospective study evaluated 190 patients across three treatment arms: rela/nivo followed by ipilimumab+nivolumab (ipi/nivo) (arm A, N=40), ipi/nivo followed by rela/nivo (arm B, N=71), and anti-PD-1 followed by rela/nivo (arm C, N=79). The study assessed the impact of treatment sequencing on outcomes including response rate, progression-free survival, and overall survival (OS). RESULTS:The overall response rate to second treatment was highest in arm C (30.4%), followed by arm B (28.1%) and arm A (17.5%). Patients with secondary resistance to first-treatment ICIs had better responses to second-treatment ICIs than those with primary resistance, particularly in arm B (p=0026). Median OS from date of first ICI treatment was significantly longer in arms B (40.9 months) and C (42.5 months) compared with arm A (16.3 months). CONCLUSIONS:Our findings indicate that rela/nivo may remain active following anti-PD-1 or ipi/nivo therapy. Additionally, our results suggest that sequencing ipi/nivo before rela/nivo may yield better outcomes than starting with rela/nivo. Patients who respond to the first combination regimen appear to derive greater benefit from the second. Further efforts are needed to optimize sequencing strategies in advanced melanoma, and future studies should consider the impact of prior treatment outcomes.

Uveal melanoma (UM) is the most common intraocular malignancy in adults. In contrast to cutaneous melanoma (CM), effective treatment options for metastatic UM are limited. The transcription factor SOX10 is crucial for CM initiation and survival, making it an interesting candidate for new targeted therapies, but its relevance in UM was unclear. We found that SOX10 was widely expressed in UM and essential for proliferation, cell cycle progression, and survival. The effects were partially mediated by SOX10-related genes including MITF , highlighting high addiction of UM to the SOX10-MITF axis. Additionally, SOX10 knockdown induced massive transcriptomic changes. Due to a lack of specific inhibitors of SOX10 and MITF, a computational approach was used to identify druggable targets by curating a UM-specific protein interaction network to search for candidates downregulated upon SOX10 inhibition. Thereby, the E2F transcription factor family was identified and their potential as druggable target candidates in UM was confirmed using the pan-E2F inhibitor HLM006474, resulting in cell cycle arrest and apoptosis. Taken together, SOX10 is crucial for UM survival and SOX10-associated proteins may serve as promising targets for developing new therapeutic strategies in UM. ### Competing Interest Statement E.K. received funding outside the submitted work from, the Bavarian Cancer Research Center (BZKF), Hiege-Stiftung die Deutsche Hautkrebsstiftung, IZKF Erlangen, Deutsche Dermatologische Gesellschaft (DDG), Arbeitsgemeinschaft Dermatologische Forschung (ADF) M. E. reports payment all outside the submitted work for lectures from: Novartis, Immunocore, and travel grant and payment for lectures from Pierre Fabre. J.V. received speakers honoraria from Novartis. C.B. reports consulting fees from BMS, Almirall Hermal, Immunocore MSD, Novartis, Regeneron, Sanofi, Pierre Fabre, Honoraria for lectures from Bristol Myers Squibb (BMS), Merck Sharpe and Dohme (MSD), Almirall Hermal, Immunocore, Novartis, Sanofi, Pierre Fabre, Leo Pharm, support for attending meetings from Pierre Fabre, and participation on advisory boards outside the submitted work of InflaRx, Miltenyi, BMS, Almirall Hermal, Immunocore, MSD, Novartis, Regeneron, Sanofi, Pierre Fabre. C.B. is a board member of the Dermatologic Cooperative Oncology group (DeCOG) M.V.H. received honoraria for lectures and presentations from Novartis BMS, MSD, Immunocore. He participated on data safety monitoring boards or advisory boards of Novartis, BMS, MSD, Immunocore. All other authors declare no conflicts of interests