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PURPOSE Mitogen-activating protein kinase inhibitors (MAPKis) are largely used in V600E/K BRAF–mutated metastatic melanomas, but data regarding effectiveness of targeted therapy in patients with rare BRAF mutations and molecular description of these infrequent mutations are scarce. PATIENTS AND METHODS A multicenter study was conducted on patients with metastatic melanoma harboring a well-identified mutation of BRAF and enrolled from March 2013 to June 2021 in the French nationwide prospective cohort MelBase. The molecular BRAF mutation pattern, response to MAPKis when applicable, and survival data were analyzed. RESULTS Of 856 selected patients, 51 (6%) harbored a non-V600E/K BRAF mutation involving codons V600 (24 of 51, 47%; V600G 27.4%, V600R 15.6%), K601 (6 of 51, 11.7%), and L597 (4 of 51, 7.8%). An objective response to MAPKis either BRAF inhibitor (BRAFi) alone or combined with MEK inhibitor was achieved in 56% (353 of 631) of V600E/K, 58% (11 of 19) of non-E/K V600, and 22% (2 of 9) of non-V600 BRAF-mutated patients, with a median progression-free survival of 7.7, 7.8, and 2.8 months, respectively. Overall, objective response rate was higher with BRAFi + MEK inhibitor combination than with BRAFi in monotherapy for each subset. CONCLUSION Rare BRAF mutations are not anecdotal in the metastatic melanoma population. Although data interpretation must remain careful owing to the limited size of some subsets of patients, non-E/K V600 BRAF mutations seem to confer a high sensitivity to targeted therapy, whereas MAPKis seem less effective in patients with non-V600 BRAF mutations. However, this strategy may be used as an alternative option in the case of immunotherapy failure in the latter population.

Proteasomes are major non-lysosomal proteolytic complexes localized in the cytoplasm and in the nucleus of eukaryotic cells. Strikingly, high levels of extracellular proteasome have also been evidenced in the plasma (p-proteasome) of patients with specific diseases. Here, we examined the process by which proteasomes are secreted, as well as their structural and functional features once in the extracellular space. We demonstrate that assembled 20S core particles are secreted by cells within microvesicles budding from the plasma membrane. Part of the extracellular proteasome pool is also free of membranes in the supernatant of cultured cells, and likely originates from microvesicles leakage. We further demonstrate that this free proteasome released by cells (cc-proteasome for cell culture proteasome) possesses latent proteolytic activity and can degrade various extracellular proteins. Both standard (no immune-subunits) and intermediate (containing some immune-subunits) forms of 20S are observed. Moreover, we show that galectin-3, which displays a highly disordered N-terminal region, is efficiently cleaved by purified cc-proteasome, without SDS activation, likely after its binding to PSMA3 (α7) subunit through its intrinsically disordered region. As a consequence, galectin-3 is unable to induce red blood cells agglutination when preincubated with cc-proteasome. These results highlight potential novel physio- and pathologic functions for the extracellular proteasome.

ObjectiveTo quantify the risk of immune-related adverse events (irAEs) in patients with pre-existing autoimmune disease (pAID) treated by immune checkpoint inhibitors (ICIs) for stage III or IV melanoma.MethodsCase–control study performed on a French multicentric prospective cohort of patients with melanoma, matched for irAE risk factors and oncological staging. Risk of irAE was assessed by logistic regression.Results110 patients with pAID were included and matched with 330 controls, from March 2013 to October 2020. Over a median follow-up period of 7.2 months for cases and 6.9 months for controls, the ORs of developing all-grade and grade ≥3 irAEs among cases compared with controls were 1.91 (95% CI (1.56 to 2.27)) and 1.44 (95% CI (1.08 to 1.82)), respectively. Patients with pAID had an increased risk of multiple irAEs (OR 1.46, 95% CI (1.15 to 2.67)) and a shorter time to irAE onset. In contrast, there were no difference in irAE-related mortality nor in the rate of treatment discontinuation, and a landmark analysis revealed a better survival at 24 months among cases (p=0.02). Thirty per cent of cases experienced a pAID flare during follow-up, and baseline immunosuppression did not prevent irAE occurrence. Last, we report associations between the pAID clinical subsets and organ-specific irAEs.ConclusionIn our study, patients with pAID were at greater risk of all-grade, severe and multiple irAEs, yet had a better 24-month survival than controls. Thus, patients with pAID should be eligible for ICI therapy but benefit from a close monitoring for irAE occurrence, especially during the first months of therapy.

Although cemiplimab has been approved for locally advanced (la) and metastatic (m) cutaneous squamous-cell carcinomas (CSCCs), its real-life value has not yet been demonstrated. An early-access program enrolled patients with la/mCSCCs to receive cemiplimab. Endpoints were best overall response rate (BOR), progression-free survival (PFS), overall survival (OS), duration of response (DOR) and safety. The 245 patients (mean age 77 years, 73% male, 49% prior systemic treatment, 24% immunocompromised, 27% Eastern Cooperative Oncology Group performance status (PS) ≥ 2) had laCSCCs (35%) or mCSCCs (65%). For the 240 recipients of ≥1 infusion(s), the BOR was 50.4% (complete, 21%, partial, 29%). With median follow-up at 12.6 months, median PFS was 7.9 months, and median OS and DOR were not reached. One-year OS was 73% versus 36%, respectively, for patients with PS &lt, 2 versus ≥ 2. Multivariate analysis retained PS ≥ 2 as being associated during the first 6 months with PFS and OS. Head-and-neck location was associated with longer PFS. Immune status had no impact. Severe treatment-related adverse events occurred in 9% of the patients, including one death from toxic epidermal necrolysis. Cemiplimab real-life safety and efficacy support its use for la/mCSCCs. Patients with PS ≥ 2 benefited less from cemiplimab, but it might represent an option for immunocompromised patients.

International audience; Background: Significant progress was recently observed in the treatment of metastatic melanoma (MM). With >50% of patients now reaching a second line of treatment and a significant improvement in the survival rate, an assessment of quality of life (QoL) during the whole course of the disease becomes necessary. The objective of this study was to describe the QoL of patients with MM in France, from their diagnosis of advanced disease to their death, in real life. Methods: QoL data were collected through MelBase, a prospective, French, multicentric cohort dedicated to the follow-up of adults with MM. QoL was assessed using the EuroQoL-5D questionnaire and the Functional Assessment of Cancer Treatment (FACT)-Melanoma questionnaire at the time of study inclusion, every 3 months, and at the time of each treatment change until death. To assess longitudinal changes from baseline to death, mixed-effect models for repeated-measures analyses were used to control for baseline covariates. Results: QoL was assessed in 1435 patients who were included in the study between 2013 and 2018. The median follow-up was 9.4 months, and 47% of patients died during follow-up. During first-line treatment, the model-based, mean utility score was 0.830 (95% CI, 0.818-0.843), the mean FACT-General score was 77.22 (95% CI, 76.23-78.22), and the mean FACT-Melanoma score was 129.46 (95% CI, 128.02-130.90). At the time of a change in treatment line, there was a decrease of −0.027 (95% CI, −0.03, −0.02) in the utility score, −1.82 (95% CI, −1.88, −1.76) in the FACT-General score, and −2.98 (95% CI, −3.05, −2.91) in the FACT-Melanoma score compared with first-line treatment. Conclusions: In the MelBase cohort, the QoL among patients with MM seems to be fairly stable over the whole disease course, although a small but significant decrease at time therapy is changed is observed.

The aim of our study was to investigate the impact of macroautophagy on exosome secretion. Exosomes are small membrane vesicles released in the extracellular space upon fusion of multivesicular endosomes with the plasma membrane. They were initially discovered as a way to remodel the reticulocyte plasma membrane before entering the blood circulation (Current Opinion in Hematology 2010, 17:177-183) and are now essentially studied as mediators of intercellular communication. Using iTRAQ proteomics, we compared the protein composition of purified exosomes secreted by cells impaired or not for macroautophagy by Atg5 depletion, during serum starvation conditions or complete medium culture. We show that the absence of serum modifies exosomal content, especially inducing secretion of two cytoplasmic protein complexes, namely proteasomal 19S regulatory particle (RP) and components of noncanonical translation preinitiation complex (PIC). This process is enhanced when autophagy is impaired by Atg5 depletion. Moreover, we show that the proteasome 20S core particle (CP) is released in the extracellular space. However, in striking contrast to what seen for its 19S RP regulator, release is independent of the exosomal vesicles, Atg5 expression and cell culture conditions. Exosome secretion can thus be considered as a cell process that participates in and reflects cell homeostasis, and care must be taken when studying potential extracellular function of exosomes due to the possible copurification of proteasome 20S CP.

Cutaneous melanoma is the most lethal type of skin cancer. Early detection is crucial to improve the outcome of melanoma patients. The identification of noninvasive prognostic biomarkers for the follow-up of melanoma patients is still in demand for clinical use. We show here that exosomal melanotransferrin fulfills the biomarker characteristics required to meet this demand. Melanotransferrin is typically overexpressed in melanoma cells compared to other cell types - including cancer cells - and is efficiently sorted and secreted with nanovesicles, or so-called exosomes, due to its membrane-anchoring by a glycosylphosphatidylinositol. Melanotransferrin is exposed on the surface of exosomes and is accessible for antibody recognition. An ELISA was set up to quantify melanotransferrin after immobilization of nanovesicles through the exosomal constituent tetraspanins CD63. Melanotransferrin was detected using a low number of exosomes purified from melanoma cell line cultures, and melanotransferrin detection was abolished by phosphatidylinositol-specific phospholipase C treatment. This exosomal melanotransferrin ELISA was able to discriminate an equal number of assayed exosomes purified from two different melanoma cell lines (A-375 vs. SK-MEL-28). Moreover, plasma samples from patients with melanoma and noncancer disease were assayed using this ELISA and elevated levels of exosomal melanotransferrin were seen in the plasma of patients with melanoma. We propose that exosomal melanotransferrin should be assessed as a potential melanoma biomarker.

9556 Background: Melanoma of unknown primary (MUP) account for 3% of all melanomas. Clinical outcome of advanced MUP in the era of novel therapies including immunotherapies (ICI) and targeted therapies (TT) have been only scarcely studied, whereas a possibly different biologic background might introduce changes in its management. Recent retrospective studies suggested that patients with advanced MUP could benefit at least as much from novel therapies as patients with known primary cutaneous melanoma (cMKP). Methods: Based on the nationwide MelBase prospective database (NCT02828202) this retrospective study included patients with advanced melanoma treated with first-line ICI, TT or CT. MUP was defined by upfront occurrence of (sub)cutaneous, nodal and/or visceral metastasis without any known prior or concomitant primary tumor. Patients with primary mucosal or ocular melanoma were excluded. Both progression-free survival (PFS) and overall survival (OS) were analyzed as co-primary variables in MUP vs cMKP, stratified by treatment subset (ICI vs TT vs CT vs whole cohort). Secondary variable was treatment-related toxicity. Multivariate analyses and propensity score analysis were performed. Objective: To investigate the efficacy and safety of systemic treatments (ICI, TT and chemotherapy (CT)) in patients with advanced MUP comparatively to stage-matched cMKP. Results: A total of 1882 patients were analyzed, including 265 (14.1%) MUP. Most patients were treated with first-line ICI. Median follow-up was 16 months. Patients in the MUP cohort more often displayed unfavorable initial prognostic factors (Table). PFS and OS did not significantly differ in MUP compared to MKP patients (p=0.73 and p=0.93 respectively). Stratification of cohorts by treatment type and application of propensity score did not lead to data modification. Treatment-related toxicity rate and severity did not differ between MUP and MKP, regardless of treatment type. Conclusions: Our results suggest that advanced MUP should be managed with similar strategies as advanced MKP. In our cohort, MUP patients benefited from novel therapies as much as MKP patients despite less favorable baseline prognostic factors. Exploratory studies investigating mutational burden and host immunity are needed to identify the underlying mechanisms. [Table: see text]

Introduction La chirurgie micrographique de Mohs (CMM) « classique » permet l’exerese des cancers cutanes avec controle anatomopathologique extemporane de 100 % des berges tumorales. Elle est peu pratiquee en France en raison de difficultes logistiques qui pourraient etre resolues en partie par une semi-automatisation de la lecture anatomopathologique. L’objectif de notre travail etait d’elaborer un algorithme d’intelligence artificielle (IA) performant capable de localiser les foyers de carcinome basocellulaire (CBC) au sein de lames histologiques de CMM via un reseau neuronal convolutif (RNC). Materiel et methodes Les lames issues des procedures CMM realisees dans notre centre entre 2016 et 2021 etaient incluses puis numerisees (grossissement x20). Les foyers de CBC etaient delimites manuellement via le logiciel Qupath. Les limites etaient systematiquement verifiees par un anatomopathologiste competent en CMM. Les images tissulaires de 71 lames issues de 49 patients etaient decoupees en patchs de 256 × 256 pixels (200 × 200 μm). Le nombre de patchs etait demultiplie (rotations, color augmentation). 106 567 patchs etaient obtenus; 7972 contenant du CBC formaient la classe « pathologique », 98595 formaient la classe « sain ». Ces patchs constituaient la base d’entrainement du RNC (Resnet-34). 10 lames issues de 10 patients differents formaient la base de test. 13 602 patchs tissulaires etaient automatiquement extraits puis classifies par le RNC pre-entraine. Premierement, on determinait un seuil de classification sain/pathologique pour que notre modele soit cliniquement performant. Puis, la qualite de superposition des foyers de CBC predits par le RNC et la verite terrain etait evaluee. Le principe de la metrique etait que chaque foyer etait considere localise si l’algorithme trouvait au moins un patch pathologique en son sein. Resultats L’AUC-ROC de notre algorithme etait de 0,9787. Avec le seuil optimal cliniquement pertinent determine a 0,21, le RNC classait correctement 12945/13602 patchs (VP= 1171, FN = 293, FP = 364, VN = 11774) soit une Se de 0,80, une Sp de 0,97, une VPP de 0,76, une VPN de 0,98. Mais les patchs classes comme FN contenaient peu de CBC (7,4 % en moyenne), ils correspondaient majoritairement a des patchs en bordure de foyer tumoral. Pour les 10 lames de test, l’approche metrique permettait de localiser correctement 212/214 foyers, (FN = 2, FP = 67). Soit une Se de 0,99 et une VPP de 0,75. Les foyers FP avaient une taille moyenne de 1,4 patchs vs. 4,3 patchs pour les VP. Discussion Par sa bonne sensibilite, notre algorithme ouvre la voie de la CMM assistee par IA. Ses performances pourront etre ameliorees en augmentant la base d’apprentissage par un recrutement multicentrique, en integrant plus de sous-types histologiques rares (sclerodermiformes) et avec les ameliorations technologiques dont beneficieront les RNC.

Introduction L’epidemie COVID-19 a contraint enseignants et etudiants a s’adapter rapidement a des modes d’enseignements « distanciels ». Le format « visioconference » s’est generalise a tous les enseignements, de la formation initiale jusqu’aux congres virtuels. Ces roues de secours, que nous pensions transitoires, ont modifie nos pratiques pendant plus d’un an : que faut-il en retenir pour notre enseignement ? Notre objectif etait d’evaluer aupres des enseignants et des internes de dermatologie, leur experience numerique au cours de l’epidemie. Materiel et methodes Entre le 10 et le 21/05/2021, une enquete par questionnaires, destinee aux enseignants et aux internes, a porte sur les thematiques suivantes: experiences en enseignement « visio »; aspects techniques; ressenti pedagogique. Les questionnaires ont ete envoyes par un lien GoogleForm aux mailing-listes « enseignants » et « internes » du CEDEF; les reponses etaient anonymes. Resultats Les taux de participation etaient de 50 repondants/106 enseignants (47 %) et de 120 repondants/453 internes (26 %). La majorite des enseignants avait participe a de multiples enseignements en « visio » (cours DFASM 96%, congres 84 %, reunion DES 70 %, cours DES 58 %, DU/DIU 56 %, EPU 60 %) et etait autonome pour la “visio” (cours DFASM et DES 78 %, reunion DES 58 %, EPU 56 %). Le logiciel Zoom etait utilise par la majorite (90 %) des enseignants et des internes. La connexion restait problematique au CHU pour plus de 80 % des repondants. La « simplicite d’utilisation » etait la qualite prioritaire d’un logiciel « visio ». Une majorite des enseignants estimait que les cours de DES (54 %) et les reunions de DES (52 %) pourraient etre perennises en distanciel, tandis que les internes estimaient que les DU/DIU (56 %), les cours de DES (57 %), les reunions de DES (56 %) et les seminaires du CEDEF (61 %) pourraient etre maintenues « en visio » apres la pandemie. Seuls 3% des internes estimaient qu’aucun cours ne devrait etre maintenu en “visio” contre 14% des enseignants (p = 0,03). Le format « hybride » des futurs congres etait plebiscite par la majorite (> 70 %) des repondants. Discussion L’experience COVID a servi de catalyseur a un nouveau mode d’enseignement, pour lequel les enseignants ont acquis une experience diversifiee et une certaine autonomie. Les enseignants sont plus reticents que les etudiants au maintien de formats « visio » en post-COVID, ces derniers privilegiant le format « visio » pour de nombreuses formes d’enseignements. Ainsi, la digitalisation de certains seminaires du CEDEF permet une accessibilite a un plus grand nombre et un suivi personnalise du parcours de l’etudiant. Le presentiel reste privilegie pour les formations post-universitaires, les solutions « hybrides » permettant de combiner les avantages des deux formats.

Metastatic melanoma is one of the most aggressive and drug-resistant cancers with very poor overall survival. Circulating melanoma cells (CMCs) were first described in 1991. However, there is no general consensus on the clinical utility of CMC detection, largely due to conflicting results linked to the use of heterogeneous patient populations and different detection methods. Here, we developed a new EPithelial ImmunoSPOT (EPISPOT) assay to detect viable CMCs based on their secretion of the S100 protein (S100-EPISPOT). Then, we compared the results obtained with the S100-EPISPOT assay and the CellSearch&reg, CMC kit using blood samples from a homogeneous population of patients with metastatic melanoma. We found that S100-EPISPOT sensitivity was significantly higher than that of CellSearch&reg, Specifically, the percentage of patients with &ge, 2 CMCs was significantly higher using S100-EPISPOT than CellSearch&reg, (48% and 21%, respectively, p = 0.0114). Concerning CMC prognostic value, only the CellSearch&reg, results showed a significant association with overall survival (p = 0.006). However, due to the higher sensitivity of the new S100-EPISPOT assay, it would be interesting to determine whether this functional test could be used in patients with non-metastatic melanoma for the early detection of tumor relapse and for monitoring the treatment response.

International audience; The prognosis of advanced melanoma has been greatly improved by new therapeutic agents and clinicians rely on dynamic signals to drive their therapeutic choices. Although the kinetics of metastatic disease seem to be correlated with survival, progression of the localized disease is not predictable.Objective: To assess whether progression of metastatic disease is associated with the time to the first distant recurrence of melanoma.Design, Setting, and Participants: This study was conducted from March 1, 2013, to September 1, 2017, among 638 adults with unresectable stage III or IV melanoma within the French multicentric prospective cohort MelBase. Patients treated with first-line immunotherapies, targeted therapies, or chemotherapy were included. Patients with unknown primary or de novo metastatic melanoma were not included. Data were analyzed from March 1, 2013, to December 1, 2017.Main Outcomes and Measures: The date of primary excision and time to first distant recurrence, progression-free survival, and overall survival were collected. Cox proportional hazards regression models were planned to assess the association between time to first distant recurrence and progression-free survival or overall survival, which was evaluated in terms of hazard ratio (HR). Time to recurrence was analyzed both as a continuous and categorical variable (24 months).Results: A total of 638 patients (272 women and 366 men; median age, 64 years [interquartile range, 52-73 years]) were included in the study. The median time from primary excision to first distant recurrence was 25 months (interquartile range, 12-55 months). There was no evidence of association of the time to recurrence with progression-free survival, both when analyzed as a continuous variable (HR, 0.99; 95% CI, 0.99-1.01) or after categorization (12-24 months: HR, 0.75; 95% CI, 0.56-1.02; >24 months: HR, 0.62; 95% CI; 0.47-1.01). There was no evidence of association of the time to recurrence with overall survival, both when analyzed as a continuous variable (HR, 0.99; 95% CI, 0.98-1.02) or after categorization (12-24 months: HR, 0.76; 95% CI, 0.54-1.07; >24 months: HR, 0.61; 95% CI, 0.54-1.03). Those results remained nonsignificant after stratification by treatment.Conclusions and Relevance: In the MelBase cohort, time to recurrence of metastatic melanoma appears not to be associated with progression-free survival or overall survival.

International audience; Background: Melanoma brain metastases (MBMs) are historically associated with poor prognosis. Radiation therapy is conventionally associated with a high local control rate. Development of targeted therapy and immunotherapy has improved overall survival (OS) and intracranial response rate, but about 50% of patients failed to respond to these novel therapies. The objective of this study was to assess the impact of combined radiotherapy (cRT) on overall survival in a large multicenter real-life prospective cohort of patients with MBM treated with immunotherapy or targeted therapy.Patients and methods: Clinical data from 262 patients with MBM were collected via MelBase, a French multicentric biobank prospectively enrolling unresectable stage III or IV melanoma. Two groups were defined: patients receiving cRT (cRT group) or not receiving cRT (no-cRT group). Primary end-point was OS. Propensity score weighting was used to correct for indication bias.Results: Among the 262 patients, 93 (35%) received cRT (cRT group). The patients were treated with immunotherapy in 69% and 60% and with targeted therapy in 31% and 40% of the cRT and no-cRT groups, respectively. With a median follow-up of 6.9 months, median OS was 16.8 months and 6.9 months in the cRT and no-cRT groups, respectively. After propensity score weighting, cRT was associated with longer OS (hazard ratio = 0.6, 95% confidence interval: 0.4-0.8; p=0.007). Median OS after ponderation was 15.3 months and 6.2 months in the cRT and no-cRT groups, respectively.Conclusion: This study shows that cRT may be associated with a significant decrease of 40% in the risk of death in patients with MBM treated with systemic therapy.

10031 Background: Obesity is an established risk factor for several cancers and higher body mass index (BMI) is associated with poor prognosis. These data are still debated in melanoma. Furthermore, recently the concept of “obesity paradox” has emerged. In a large cohort published by McQuade JL et al, higher BMI was associated with better survival in patients with metastatic melanoma (MM) especially for those treated with targeted therapy (TT) and immune checkpoint inhibitors (ICI). We studied the association between BMI and progression-free survival (PFS) and overall survival (OS) in patients with MM treated with systemic therapies. Methods: This study was conducted from the prospective MelBase cohort (NCT02828202). Patients with MM treated with first-line ICI, TT, or CT were included. BMI was categorized by WHO criteria. Underweight patients were excluded. The co-primary outcomes were the associations between BMI and PFS or OS, stratified by treatment, sex and age. Multivariate analyses were performed. Results: A total of 1214 patients were analyzed. The majority of them were treated with ICI, followed by TT. Obese patients represented 22% of cohort (Table). Median follow-up was 13.5 months. The patients who were overweight or obese did not have different PFS (p = 0.88) or OS (p = 0.25) than patients with normal BMI. Stratifying this cohort by treatment received, age, sex and others parameters (such as LDH, number of metastatic site) did not revealed any difference. Multivariate analysis did not change the results. Conclusions: BMI was not associated with clinical outcomes in our cohort, especially in ICI and TT groups. Thus, we did not confirm the results presented by McQuade Jl et al. with a cohort quite similar in term of size. Because BMI is too simplistic and then an imperfect measure of body composition, the published data are not reproducible. We caution the oncologists, about BMI as valuable predictive marker of survival for melanoma patients. [Table: see text]

International audience; Rarely, basal cell carcinomas (BCCs) have the potential to become extensively invasive and destructive, a phenomenon that has led to the term "locally advanced BCC" (laBCC). We identified and described the diverse settings that could be considered "locally advanced". The panel of experts included oncodermatologists, dermatological and maxillofacial surgeons, pathologists, radiotherapists and geriatricians. During a 1-day workshop session, an interactive flow/sequence of questions and inputs was debated. Discussion of nine cases permitted us to approach consensus concerning what constitutes laBCC. The expert panel retained three major components for the complete assessment of laBCC cases: factors of complexity related to the tumour itself, factors related to the operability and the technical procedure, and factors related to the patient. Competing risks of death should be precisely identified. To ensure homogeneous multidisciplinary team (MDT) decisions in different clinical settings, the panel aimed to develop a practical tool based on the three components. The grid presented is not a definitive tool, but rather, it is a method for analysing the complexity of laBCC.

International audience; The need for bioactive dressings increases with the population aging and the prevalence of chronic diseases. In contrast, there are very few dressings on the market which are designed to display a chosen bioactivity. In this context, we investigated the surface-functionalization of silicone wound dressing with bioactive peptides. One of the challenges was to avoid multistep grafting reactions involving catalysts, solvents or toxic reagents, which are not suitable for the fabrication of medical devices at an industrial scale. In the other hand, a covalent bonding was necessary to avoid the loss of the biological effect by progressive removal of the peptide in biological fluids generated by the wound. To solve these limitations, we developed a strategy allowing an easy and direct functionalization of silicone. This strategy relies on hybrid silylated bioactive peptides, which chemoselectively react with plasma-activated silicone surfaces. We synthesized three hybrid peptides with wound healing properties, which were grafted on commercially available silicone dressings Cerederm® and Mepitel®. Grafted dressings were evaluated in vitro and enabled a quicker scare recovery and extracellular matrix deposition with human dermal fibroblasts. These results were confirmed by in vivo studies showing an enhanced wound-healing of the pig skin. By this simple method, we transformed inert dressing into bioactive dressing which showed properties of wound healing.

Background: Macular arteritis, macular lymphocytic arteritis (MLA) or lymphocytic thrombophilic arteritis all correspond to an identical new clinicopathological entity. Its individualization as a primary cutaneous lymphocytic arteritis is still controversial for certain authors as it could represent a latent form of cutaneous polyarteritis nodosa. Materials and Methods: We report here 3 additional cases of MLA, present a review of the literature and discuss the disease's nosology. Results: MLA is characterized clinically by a benign skin eruption consisting in bilateral asymptomatic erythematous/hyperpigmented macules mainly located on the lower legs and histologically by a medium-sized cutaneous lymphocytic prominent arteritis present in early cutaneous lesions. Conclusion: These findings support that MLA may be considered as a chronic and indolent primary lymphocytic cutaneous arteritis. Nevertheless, in some cases the objective obliteration of cutaneous vessels underlines the need for continuous monitoring in MLA patients.

Plasmatic proteasome (p-proteasome) has recently been described as a new marker for metastatic melanoma. The objective of this study was to compare the diagnostic and prognostic values of p-proteasome with three other melanoma serological markers: S100B protein, melanoma inhibitory activity protein (MIA) and lactate dehydrogenase (LDH) in the plasma of 121 stage I-IV melanoma patients. Laboratory analyses were performed by standardized ELISA (p-proteasome, MIA), immunoluminometric assay (S100B) and colorimetry (LDH). We found that all markers were relevant for discriminating metastatic from nonmetastatic patients but p-proteasome displayed the highest diagnostic accuracy. P-proteasome and S100B were the most sensitive (58.1%) and p-proteasome and MIA the most specific (98.7 and 100%) in detecting metastatic disease. P-proteasome and S100B had the highest area under receiver operating characteristics curve, 0.811 (95% CI: 0.725-0.897) and 0.822 (95% CI: 0.738-0.906), respectively. These two markers were the best in detecting patients with lymph node metastases. S100B, MIA and LDH diagnostic accuracy was increased when these markers were combined with p-proteasome. As shown with univariate analysis, shorter progression-free and overall survival rates were significantly associated with elevated plasma levels of each markers. The multivariate Cox regression analysis identified p-proteasome as the only independent predictor of a poorer progression-free survival (p = 0.030). In conclusion, this comparative study established that p-proteasome quantification in combination with other melanoma biomarkers is an attractive approach for the biological follow-up of melanoma patients.

Background The ubiquitin proteasome pathway is involved in the pathogenesis of psoriasis and proteasome subunits are increased in lesional psoriatic skin. Recent works have highlighted that proteasome levels can be regulated through modulation of proteasome assembly notably by the proteasome maturation protein POMP. Objectives To investigate whether proteasome assembly and POMP expression are modified in psoriatic skin. Methods Proteasome assembly as well as expression of proteasome regulators were assessed in non-lesional and lesional psoriatic skin using native gel electrophoresis and western blots respectively. The protein and mRNA expression levels of POMP were compared by western blots, immunohistochemistry and quantitative polymerase chain reaction. The role of POMP in keratinocyte proliferation and differentiation was assessed by silencing POMP gene expression by RNA interference in human immortalized keratinocyte HaCaT cells. Results Both 20S and 26S proteasomes (and their respective proteolytic activities) as well as the main proteasome regulators are increased in lesional psoriatic skin. POMP binds to 20S precursor complexes and is overexpressed in lesional epidermal psoriatic skin, supporting that POMP-mediated proteasome assembly is increased in psoriatic skin. POMP silencing inhibited HaCaT cell proliferation and induced apoptosis through the inhibition of the proteasome assembly. Moreover POMP partial depletion decreased the expression of the differentiation markers keratin 10 and involucrin during the [Ca 2+ ]-induced HaCaT cells differentiation. Conclusion Altogether these results establish a potential role for POMP and proteasome assembly in psoriasis pathogenesis.

Plasmatic proteasome (p-proteasome) also called circulating proteasome has recently been described as a tumor marker. We investigated the diagnostic and prognostic accuracies of p-proteasome levels in a melanoma population classified according to the American Joint Committee on Cancer staging system. Using an ELISA test, we measured p-proteasome levels in 90 patients and 40 controls between March 2003 and March 2008. The subunit composition of p-proteasomes was determined in metastatic melanoma by proteomic analysis. The mean p-proteasome levels were correlated with stages (P 4300 ng/ml) were significantly correlated with an increased risk of progression [hazard ratio (HR) = 7.34; 95% CI 3.54-15.21, P < 0.0001] and a risk of death (HR = 5.92; 95% CI 2.84-12.33, P < 0.0001). In multivariate analysis, high p-proteasome levels were correlated with a poorer clinical outcome in the subgroup analysis limited to patients with disease stages I, II and III. Proteomic analysis confirmed the presence of all proteasome and immunoproteasome subunits. Taken together, these results indicate that p-proteasomes are a new marker for metastatic dissemination in patients with melanoma.

e21536Background: Melanoma’s incidence is increasing, and elderly people could be significantly impacted since more than 50% of melanoma arise in > 65 years-old (yo). Combined BRAF and MEK targeted...

Alpha-melanocyte stimulating hormone (alpha-MSH) binds to melanocortin-1 receptor (MC1R) on melanocytes to stimulate pigmentation and modulate various cutaneous inflammatory responses. MC1R expression is not restricted to melanocytic cells and may be induced in keratinocytes after UVB exposure. We hypothesized that MC1R signaling in keratinocytes, wherein basal conditions are barely expressed, may modulate mediators of inflammation, such as nuclear factor-kappa B (NF-kappaB) and tumor necrosis factor-alpha (TNF-alpha). Therefore, we generated HaCaT cells that stably express human MC1R or the Arg151Cys (R151C) nonfunctional variant. We demonstrate that: (1) the constitutive activity of MC1R results in elevated intracellular cAMP level, reduced NF-kappaB activity and decreased TNF-alpha transcription; (2) binding of alpha-MSH to MC1R and the subsequent increase in cAMP production do not inhibit TNFalpha-mediated NF-kappaB activation; (3) MC1R signaling is sufficient to strongly inhibit UVB-induced TNF-alpha expression and this inhibitory effect is further enhanced by alpha-MSH stimulation. Our findings suggest that the constitutive activity of the G-protein-coupled MC1R in keratinocytes may contribute to the modulation of inflammatory events and immune response induced by UV light.

International audience; IL-20 is involved in the development of skin psoriasis. The molecular mechanisms underlying IL-20 overexpression in psoriatic epidermis remain to be elucidated. We showed that IL-20 was primarily upregulated in psoriatic skin at the post-transcriptional level. The RNA-binding protein HuR relocalized to the cytoplasm of keratinocytes (KCs) of psoriatic patients, suggesting that it stabilizes numerous transcripts, as observed in the human KC cell lines used to assess IL-20 mRNA. We characterized epidermal HuR RNA targets in psoriatic skin using ribonucleoprotein immunoprecipitation analyzed via high-throughput sequencing. Numerous transcripts that are upregulated in psoriasis were targeted by HuR, supporting the participation of HuR in pathogenic processes such as morphological changes, innate and adaptive immune responses, and metabolic inflammatory responses. Finally, we identified the metabolic sensor AMP-activated protein kinase (AMPK) as being responsible for HuR cytoplasmic relocalization because its activity was severely impaired in human psoriatic epidermis, and in vivo drug-mediated AMPK inhibition in mouse epidermis promoted HuR cytoplasmic localization, IL-20 overproduction, acanthosis, and hyperkeratosis. These results provide insights into the molecular links between metabolism and post-transcriptional networks during chronic inflammation.

International audience; BACKGROUND:Loxoscelism is an envenomation due to a bite by spiders of the genus Loxosceles, very well known on the American continent but unrecognized in Europe.CASE REPORT:We report the case of a 36-year-old woman, without any medical history or treatment, who went to a University Hospital in the South of France, for a painful skin lesion on the internal part of her left thigh, which appeared in the morning and developed rapidly during the day. She was directed to the infectious disease department with a diagnosis of skin infection. In spite of the antibiotics, the lesion increased, with a hemorrhagic central blister, an irregular ecchymotic center, a pale perimeter, and an extensive inflammatory and indurate oedema affecting the whole thigh. There was also a low-grade fever, chills, intense pain and a generalized scarlatiniform exanthema. The lesion was finally diagnosed as cutaneous loxoscelism, then confirmed by collection and identification of a Loxosceles rufescens spider killed by the patient the morning of the occurrence of the lesion. Following an initial symptomatic treatment, the development of a necrotic ulcer justified a delayed surgical reconstruction, after stabilization of the lesion.CONCLUSIONS:Loxosceles bites are usually painless and rarely noticed by patients, often leading to a presumptive diagnosis. Therefore, in the case of a dermonecrotic lesion developing unfavourably with antibiotics, cutaneous loxoscelism should be one of the diagnoses to be considered.

Introduction La detection des cellules tumorales circulantes (CTCs) peut etre importante dans la prise en charge des malades atteints de melanomes metastatiques (MMM). Les CTCs sont heterogenes et seules les CTCs viables sont responsables de la dissemination metastatique ; peu de methodes permettent actuellement de les detecter. L’objectif de notre travail a ete d’evaluer par une methode innovante (EPIthelial immunoSPOT ou EPISPOT) le nombre de CTCs vivantes chez des MMM et de comparer les resultats obtenus avec ceux de la technique de reference (CellSearch® ou CS). Materiel et methodes Cette etude prospective, comparative, monocentrique a ete realisee chez 35 MMM (stade IV AJCC) et 20 volontaires sains. Le nombre de CTCs chez chaque sujet a ete determine en aveugle a l’aide des deux methodes. Pour chaque prelevement, un controle interne a ete realise avec des cellules provenant de la lignee WM-266-4. Le marqueur proteique retenu pour l’analyse par EPISPOT a ete la proteine S100B. Apres depletion des cellules CD45+, les prelevements sanguins ont ete deposes sur une membrane de nitrocellulose recouverte d’anticorps anti-PS100. Apres 48 heures d’incubation, les immunospots ont ete comptes a l’aide d’un microscope a fluorescence, chacun d’eux correspondant a une CTC viable secretrice de PS100B. Les marqueurs CD146, HMWMAA, DAPI, CD45 et CD34 ont ete utilises pour definir les CTCs par le systeme CS. Resultats Le systeme CS a permis de detecter au moins 1 CTC chez 31,4 % (11/35) des MMM et au moins 2 CTCs chez 20 % d’entre eux (7/35). La technique EPISPOT a permis de detecter au moins 1 CTC viable chez 51,4 % (18/35) des MMM et au moins 2 CTCs chez 40 % d’entre eux (14/35). Avec un cut-off standard ≥ 2 CTCs, le systeme EPISPOT avait une meilleure sensibilite que le systeme CS (p = 0,035). Une mediane de 1 CTC (etendue 0–900) a ete detectee par EPISPOT et de 0 CTC (etendue 0–4937) par le systeme CS dans le groupe MMM. Il n’y avait pas de difference statistiquement significative dans la specificite des deux methodes. Discussion La technique EPISPOT est basee sur la secretion de marqueurs proteiques specifiques par les CTCs apres une courte phase de culture cellulaire. Elle a deja ete utilisee pour la detection des CTCs fonctionnelles dans les cancers du sein, du colon et de la prostate. A notre connaissance, elle n’a jamais ete proposee dans le melanome. Les resultats de notre etude montrent qu’elle a une sensibilite superieure a celle de la methode immunomagnetique de reference. Conclusion Il s’agit de la premiere etude evaluant la technique EPISPOT dans le melanome. L’EPISPOT PS100, base sur les proprietes fonctionnelles de CTCs viables, a une meilleure sensibilite que celle du systeme immunomagnetique de reference (CS) qui ne repose que sur le phenotype des CTCs. Son utilisation pourrait etre proposee dans la detection des CTCs chez les malades atteints de melanome metastatique.

9551 Background: With increasing armamentarium in advanced melanoma management, the impact of various strategies remains to be determined including the importance of time to switch from one treatment to another. We report the impact of time to IPI/APD non-planned switch on APD efficacy in real life patients within MelBase (MB). Methods: MB is a French multicentric biobank dedicated to the prospective follow-up (FU) of unresectable stage III or IV melanoma with 1102 patients included since March 2013. Data were collected (Sept.2016) and analyzed (demography, overall survival (OS), progression-free survival (PFS), response rate, multivariate analysis, safety). Results: 71 patients were treated with IPI/APD sequence. 72% received 4 IPI injections. The median time to switch was 1.7 months (0.36-3). The characteristics at the initiation of APD are: mean age 64 yrs, PS 0-1 80%, elevated LDH 34%, BRAF WT 90%, brain metastasis 25%, ≥ 3 metastatic organ sites (MOS) 49%, median FU 11.9 months, OS 20 months (95%CI:12.6-NR), PFS 3.5 months (95%CI:2.9-6.2). The best overall response was 25%, disease control rate was 54% with a low toxicity profile (17% grade 3/4). In a multivariable analysis, longer time to switch was significantly associated with better OS (adjusted HR 0.38 per 1 more month, 95%CI:0.14-0.93), as well as ECOG 0-1 (aHR 3.11, 95%CI:0.99-9.72) and LDH < ULN (aHR 3.32, 95%CI:1.26-8.75). In addition, the association of time to switch with OS vary significantly according to the number of MOS ( < 3 MOS aHR 0.25, 95%CI:0.10-0.62; ≥3 MOS aHR 0.99 95%CI:0.41-2.39) and AJCC stage (M0/1a/1b aHR 0.06, 95%CI:0.01-0.43 ; M1c aHR 0.77, 95%CI:0.39-1.54). Conclusions: In patients who failed IPI treatment, longer survival after APD was associated to time to switch only in patients with favorable baseline factors. Such results are probably more related to the slow kinetics of the disease than to the delay itself. Our results are different from Blank et al.(ESMO 2016) who tested a planned switch, immediately after 2 IPI perfusions, and showed overall response rate close to IPI+APD association. We are currently conducting a similar study on the reverse sequence (APD/IPI) and the role on IPI efficacy.

9538 Background: In the past decade, the prognosis of advanced melanoma has been greatly improved by new therapeutic agents such as immune and targeted therapies, which are now being evaluated as adjuvant therapies. Although the kinetic of metastatic disease is correlated to patient survival, the unfolding of the dormant disease remains hardly predictable and data from the literature on the topic is controversial (Karakousis, Francken). We hypothesized that the course of the advanced disease could be predicted from time to distant recurrence. Methods: Melbase is a French multicentric cohort dedicated to the prospective follow-up of adults with unresectable stage III or IV melanoma. Date of primary excision, time to recurrence, progression free survival (PFS) and overall survival (OS) of 298 patients treated in first line by immune therapies (IT, n = 148), targeted therapies (TT, n = 68) or within clinical trials (n = 73) were collected on September 5th, 2016. Patients with unknown primary or de novo metastatic melanoma were not included. Time to distant recurrence was analyzed as a continuous variable and as a categorical variable ( < 12 months; 12 to 24 months; ≥ 24 months). Results: Patients’ characteristics at baseline are: mean age 62 years, PS 0-1 84%, elevated LDH 32%, BRAF mutated 39%, brain metastases 18%. Time to recurrence studied as a continuous variable was not correlated to PFS (HR = 0.96; 95%CI: 0.85-1.07) or OS (HR = 0.89; 95%CI: 0.77-1.03). These results remained insignificant after stratification upon treatment or even when time to recurrence was analyzed as a categorized variable. Conclusions: Our data showed no correlation between the time from primary to distant recurrence and PFS or OS in patients treated with TT or IT. Therefore, kinetics of advanced disease cannot be predicted by the history of dormant disease. Dormancy and metastasis proliferation are thus probably driven by different molecular and cellular mechanisms.

Introduction L’activite proteolytique et l’expression des sous-unites des proteasomes sont augmentees dans l’epiderme psoriasique. Nos precedents resultats etaient en faveur d’une regulation post-transcriptionnelle de cette expression. Notre objectif etait d’etudier l’assemblage du proteasome et l’expression d’une proteine chaperonne cle de l’assemblage du proteasome, la proteine de maturation du proteasome (POMP), dans la peau psoriasique et au cours de la differenciation keratinocytaire. Materiel et methodes Les expressions des proteasomes et de POMP etaient evaluees par IHC, WB et RT-PCR. L’assemblage du proteasome etait etudie par electrophorese PAGE (conditions non denaturantes) a partir de lysats de peaux psoriasiques ou de keratinocytes. L’expression de POMP etait modulee avec des ARN interferents dans des lignees keratinocytaires HaCat. Resultats Nous montrons que tous les sous-types (30S, 26S, 20S) et tous les regulateurs (PA28, PA200, 19S) des proteasomes sont surexprimes dans la peau lesee ou les proteasomes 26S sont proportionnellement plus augmentes que les 20S (en faveur d’un assemblage accru) et ou POMP est surexprimee et preferentiellement associee a des precurseurs du proteasome. L’inhibition de POMP a un effet anti-proliferatif et pro-apoptotique via l’inhibition de l’assemblage du proteasome dans les lignees keratinocytaires. Cette inhibition diminue l’expression de keratine 10 et d’involucrine au cours de la differenciation calcium-induite des HaCat. Discussion Ces resultats indiquent que l’augmentation de l’assemblage, de l’activite et de la quantite des proteasomes dans l’epiderme psoriasique lese est liee (au moins en partie) a la surexpression de POMP. De plus, POMP peut reguler la proliferation et la differenciation keratinocytaire. Conclusion Un ciblage therapeutique de POMP en agissant a la fois sur les proteasomes et les immunoproteasomes aurait une action anti-inflammatoire et anti-proliferative potentiellement interessante et adaptee au traitement du psoriasis.

Introduction Les kinases dependantes des cyclines (Cdk) sont actives si elles forment des complexes avec les cyclines. Les Cdk4/Cycl D1 et Cdk2/Cyc E interviennent dans la regulation du cycle cellulaire : sortie de quiescence et progression a travers la phase G1 pour Cdk4/Cyc D1, transition G1/S pour Cdk2-Cyc E. Notre objectif a ete d’evaluer, dans l’epiderme humain normal et psoriasique, l’expression et l’activite fonctionnelle des Cdk4 et Cdk2 grâce a des biosenseurs cutanes specifiques. Materiel et methodes Notre etude a porte sur 20 malades atteints de psoriasis moderes a severes. Chez chaque patient, des biopsies ont ete pratiquees en peau lesee et en peau saine perilesionnelle. L’expression des molecules d’interet a ete etudiee par immunohistochimie, microscopie confocale et immunoblots epidermiques. La transcription des genes a ete evaluee par qPCR et l’activite fonctionnelle des Cdk4 et Cdk2 a ete quantifiee a l’aide de biosenseurs fluorescents specifiques (WWH3c pour Cdk2, WwshortRbc pour Cdk4) en presence ou des non-inhibiteurs de Cdk2 (roscovitine) et de Cdk4 (abemaciclib). Resultats Il existait une induction de l’expression cytoplasmique des Cdk2 et Cdk4 dans l’epiderme psoriasique avec conservation de celle-ci dans les cellules les plus differenciees. Les immunoblots epidermiques permettaient de confirmer l’induction de Cdk4 et la forte augmentation de Cdk2. Les resultats de l’analyse en qPCR ont montre une discrete inhibition des genes codant pour Cdk4 et une nette inhibition de la transcription de Cdk2. L’activite fonctionnelle de Cdk2 etait augmentee dans l’epiderme lesionnel contrairement a celle de la Cdk4 qui etait effondree. Cette inhibition de Cdk4 en peau lesee etait associee a une augmentation de l’expression cytoplasmique et nucleaire de la cyc D1 et de la proteine p21 en peau lesee. Il existait, en revanche, une diminution de l’expression membranaire de la proteine p27 dans l’epiderme psoriasique. On retrouvait une forte augmentation de l’expression de la proteine p16 (inhibiteur de Cdk4) dans l’epiderme lesionnel. Discussion Il existe in vivo dans l’epiderme psoriasique un effondrement de l’activite fonctionnelle des complexes Cdk4/Cyc D1. Cette inhibition ne resulterait pas d’une diminution de l’expression des proteines Cdk4 et cyc D1 qui sont au contraire fortement exprimees en peau lesee. Elle pourrait etre secondaire a une inhibition de leur degradation par le proteasome. L’effondrement de l’activite des Cdk4 pourrait etre due a une sequestration de ces proteines ou a l’augmentation de leurs inhibiteurs physiologiques p16 et p21. L’augmentation de l’activite des complexes Cdk2/cyc E pourrait resulter de l’augmentation d’expression de la Cdk2. Elle pourrait etre egalement due a une levee d’inhibition par p27 dont l’expression est diminuee dans l’epiderme psoriasique. Conclusion Ces resultats permettent de mieux comprendre les mecanismes de l’homeostasie epidermique au cours du psoriasis.

9556Background: Melanoma brain metastases (BM) are associated with poor prognosis. Although novel therapies such as ipilimumab (IPI) and anti-PD1 (aPD1: nivolumab and pembrolizumab) have shown thei...

Plasmatic proteasome (p-proteasome) has recently been described as a new marker for metastatic melanoma. The objective of this study was to compare the diagnostic and prognostic values of p-proteasome with three other melanoma serological markers: S100B protein, melanoma inhibitory activity protein (MIA) and lactate dehydrogenase (LDH) in the plasma of 121 stage I-IV melanoma patients. Laboratory analyses were performed by standardized ELISA (p-proteasome, MIA), immunoluminometric assay (S100B) and colorimetry (LDH). We found that all markers were relevant for discriminating metastatic from nonmetastatic patients but p-proteasome displayed the highest diagnostic accuracy. P-proteasome and S100B were the most sensitive (58.1%) and p-proteasome and MIA the most specific (98.7 and 100%) in detecting metastatic disease. P-proteasome and S100B had the highest area under receiver operating characteristics curve, 0.811 (95% CI: 0.725-0.897) and 0.822 (95% CI: 0.738-0.906), respectively. These two markers were the best in detecting patients with lymph node metastases. S100B, MIA and LDH diagnostic accuracy was increased when these markers were combined with p-proteasome. As shown with univariate analysis, shorter progression-free and overall survival rates were significantly associated with elevated plasma levels of each markers. The multivariate Cox regression analysis identified p-proteasome as the only independent predictor of a poorer progression-free survival (p = 0.030). In conclusion, this comparative study established that p-proteasome quantification in combination with other melanoma biomarkers is an attractive approach for the biological follow-up of melanoma patients.

International audience; Ultraviolet A (UVA) radiations are responsible for deleterious effects, mainly due to reactive oxygen species (ROS) production. Alpha-melanocyte stimulating hormone (α-MSH) binds to melanocortin-1 receptor (MC1R) in melanocytes to stimulate pigmentation and modulate cutaneous inflammatory responses. MC1R may be induced in keratinocytes after UV exposure. To investigate the effect of MC1R signaling on UVA-induced ROS (UVA-ROS) production, we generated HaCaT cells that stably express human MC1R (HaCaT-MC1R) or the Arg151Cys (R(151)C) non-functional variant (HaCaT-R(151)C). We then assessed ROS production immediately after UVA exposure and found that: (1) UVA-ROS production was strongly reduced in HaCaT-MC1R but not in HaCaT-R(151)C cells compared to parental HaCaT cells; (2) this inhibitory effect was further amplified by incubation of HaCaT-MC1R cells with α-MSH before UVA exposure; (3) protein kinase A (PKA)-dependent NoxA1 phosphorylation was increased in HaCaT-MC1R compared to HaCaT and HaCaT-R(151)C cells. Inhibition of PKA in HaCaT-MC1R cells resulted in a marked increase of ROS production after UVA irradiation; (4) the ability of HaCaT-MC1R cells to produce UVA-ROS was restored by inhibiting epidermal growth factor receptor (EGFR) or extracellular signal-regulated kinases (ERK) activity before UVA exposure. Our findings suggest that constitutive activity of MC1R in keratinocytes may reduce UVA-induced oxidative stress via EGFR and cAMP-dependent mechanisms.

Ultraviolet (UV) exposure induces an up-regulation of melanocortin-1 receptor (MC1R) expression in human skin and the alpha-melanocyte-stimulating hormone (alpha-MSH) may reduce UVB-induced DNA damage in normal human melanocytes. Using high-performance liquid chromatography coupled to tandem mass spectrometry, we investigated the formation and repair of DNA lesions in UVB-irradiated HaCaT cells stably transfected with the wild type MC1R gene (HaCaT-MC1R). Similar levels of 8 bipyrimidine photoproducts including cyclobutane pyrimidine dimers (CPDs) (TT, TC, CT), (6-4) photoproducts ((6-4)PPs) (TT-(6-4)PPs, TC-(6-4)PPs) and their Dewar valence isomers together with 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) were found to be generated in both non-transfected and HaCaT-MC1R cells after UVB exposure. Time-course studies of DNA photoproduct yields indicated that the DNA repair ability depended upon radiation doses. It was shown that (6-4)PPs were removed from the DNA of UVB-irradiated cells much more efficiently than CPDs. The repair efficiency of 8-oxodGuo, CPDs and (6-4)PPs was relatively similar in both cell lines and was not modified by stimulation with alpha-MSH before UVB-exposure. In conclusion, cell surface-enforced expression of MC1Rs on HaCaT keratinocytes and alpha-MSH stimulation do not affect the formation of UVB-induced DNA photoproducts and their subsequent repair.

BACKGROUND Kaposi sarcoma (KS) is an angioproliferative disease that may represent a difficult therapeutic challenge in disseminated stages. The efficacy of taxanes (paclitaxel and docetaxel), as agents with antiangiogenic properties, has been described previously in the treatment of patients with acquired immunodeficiency syndrome (AIDS)-associated KS but remains unknown in the treatment of patients with refractory or life-threatening KS without human immunodeficiency (HIV) infection. METHODS During the past 6 years, 12 non-HIV-infected patients with refractory KS were treated with paclitaxel (175 mg/m2 every 3 wks) or docetaxel (60 mg/m2 every 3 wks). RESULTS All patients improved dramatically after chemotherapy. Partial desinfiltration (n = 6) or complete desinfiltration (n = 6) of all papulonodular skin lesions was observed with marked improvement of lymphedema in 6 patients. According to the AIDS Clinical Trials Group criteria, response was partial in 100% of patients. A major response was obtained among patients who had visceral lesions (n = 3 patients), with rapid and complete remission of digestive or respiratory symptoms. The mean delay to response was 2 courses. Treatment was sustained for 3 patients. For the remaining 9 patients who received a median of 7 courses (range, 2-14 courses), the mean time to recurrence or follow-up without recurrence was 13 months. Tolerance was good except for 3 episodes of Grade 3 or 4 asymptomatic neutropenia and 1 episode of moderate myositis. CONCLUSIONS The results of this study showed that taxanes are beneficial in the treatment of patients with refractory or life-threatening Kaposi sarcoma. Cancer 2006. © 2006 American Cancer Society.

Ross syndrome is a rare partial dysautonomic syndrome of unknown aetiology, characterized by a one-sided or bi-lateral anhidrosis associated with Holmes-Adie syndrome (defined by tonic pupil and hyporeflexia of deep tendons) (1). Since it was first described in 1958 (2), approxima-tely 50 cases have been described in the literature (3). Dermatologists may be confronted with this disorder of thermoregulation, which is often revealed by a segmental “compensatory” hyperhidrosis. We describe here a case of Ross syndrome associated with Sjogren syndrome (SS) and discuss the potential relationship between these two entities. Thermography imaging exploration with an in-frared camera performed in different ambient temperature settings allowed better definition of the thermoregulatory sweating abnormalities in our patient. Moreover, to our knowledge, this examination showed, for the first time, an unexpected body “thermal pattern”, which may be characteristic of Ross syndrome. CASe RepoRTWe present here a case of a 35-year-old woman with a 10-year medical history of localized hyperhidrosis. Her medical history revealed autoimmune thyroid disease and SS. The hyperhidrosis appeared at an ambient tem-perature of approximately 25–27°C and was localized on the left side of her lower back, extending belt-like towards the side and in the controlateral lower back on the right side, as well as on the nape of her neck and shoulders. The rest of the skin was anhidrotic. She was also heat intolerant, with lack of sweating, abnormal increase in body temperature and generalized fati-gability when exposed to high ambient temperatures (32–34°C). Both pupils showed a slow reaction to light and a normal constriction to a near target, and deep tendon jerks were absent without motor deficits (Holmes-Adie syndrome). There were no other signs of autonomic dysfunction. punch biopsies obtained from hyperhidrotic and hypohidrotic skin areas were normal. The skin direct immunofluorescence test was negative. Blood explorations showed an isolated presence of anti-SSA, anti-SSB antibodies and anti-thyroid antibodies. Serum ganglionic (AChR) antibody was negative. Oral and conjunctival Sicca were objectively confirmed by Schirmer’s test. Salivary gland biopsy revealed lympho-cytic infiltration in both periglandular and perivascular regions. All of these Sicca findings fulfilled the diag-nostic criteria for primary SS (4). Brain and spinal cord magnetic resonance imaging (MRI), as well as thoracic and abdominal computed tomography scan findings, were normal. Electrophysiological explorations of both hands and forearms showed an absence or collapse of the sympathetic skin responses. Nerve conduction study findings, EEG and cardiac tilt-test (reflecting the adren-ergic sympathetic and parasympathetic functions) were normal. Thermographic explorations with an infrared camera (A320, FLIRsystems, Inc., Issy Les Moulineaux, France) allowed for evaluating the body’s surface tem-perature at 22°C and after a 20 min exposure to 32°C ambient temperature (Fig. 1). This examination was significantly more efficient than the Minor Starch Test and clearly delimited the skin thermoregulation abnor-malities associated with Ross syndrome. Hyperhidrotic and anhidrotic areas were also clearly visualized as the lowest and highest estimated skin surface temperature. This examination showed large dermatomal-like pat-terns in hyperhidrotic areas (lower back and shoulder regions), but also detected unexpected smaller patch

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e20113 Background: Recent advances in treatment of metastatic melanoma highlight how important collecting and building databases are, for a better health care management (overall survival, safety, ...

Contexte : le lichen scléreux vulvaire (LSV) est une pathologie d’étiologie inconnue entraînant un prurit, des douleurs, une atrophie et sclérose cutanée et favorisant le développement de carcinome épidermoïde vulvaire. Les corticoïdes locaux appliqués au long cours sont le seul traitement recommandé, mais exposent à long terme au risque d’atrophie cutanée et de surinfection. La photobiomodulation (PBM) est un traitement bien toléré qui a montré de bons résultats dans plusieurs pathologies dermatologiques et dans le syndrome génito-urinaire post-ménopausique. Le but de notre étude préliminaire était de tester la faisabilité, la tolérance et l’efficacité de la PBM dans le LSV.Méthodes : l’étude a porté sur 10 patientes. Le protocole thérapeutique de PBM (MILTA, MILTA-PHYSIOQUANTA, France) était celui habituellement utilisé pour traiter l’atrophie génitale ménopausique : les lésions de LSV étaient exposées à 6 séances de PBM de 20 minutes chacune, émettant des rayonnements laser NLPC, infrarouge, magnétique et LED, à la fréquence de 2 séances par semaine. L’efficacité du traitement et sa tolérance étaient évaluées par des scores basés sur les signes fonctionnels (SSF), les signes physiques (SSP), un score de qualité de vie (DLQI) avant PBM, un mois (M1) et 4 mois (M4) après la fin du traitement.Résultats : sept patientes étaient atteintes d’un LSV sévère. Aucune douleur ou sensation désagréable n’était rapportée lors du traitement. Les SSF, SSP et le DLQI moyens (13 +/- 3.64, 4.9 +/- 0.7, 8.3 +/- 2.23 respectivement) ont tous diminué à M1 (9.9 +/- 2.15, 2.9 +/- 0.8, 5.3 +/- 1.4 respectivement) et M4 (8.9 +/- 2.6, 4.3 +/- 1, 5.7 +/- 1.4 respectivement). Seule la diminution du SSP moyen à M1 était statistiquement significative (p = 0.02). Six patientes ont pu interrompre plusieurs semaines les corticoïdes locaux. L’atrophie génitale cutanéo-muqueuse était le signe clinique le plus souvent amélioré (n = 8 et n = 5 respectivement à M1 et M4). Quatre patientes sur dix présentaient une augmentation de leur SSF à M1 et/ou ré-introduisaient la corticothérapie locale avant M1. La ré-augmentation des scores d’évaluation moyens entre M1 et M4 suggérait une efficacité transitoire du protocole thérapeutique utilisé.Conclusion : nous rapportons la faisabilité et l’intérêt de la photobiomodulation dans le traitement du LSV. Ces résultats devront être confirmés par une étude de plus grande ampleur et par un essai randomisé comparatif vs corticothérapie en double aveugle. Un protocole thérapeutique adapté au LSV intégrant des irradiations initiales plus nombreuses et un traitement d’entretien pourrait également être évalué.