MAPKinase inhibition after failure of immune checkpoint blockade in patients with advanced melanoma – an evaluation of the multicenter prospective skin cancer registry ADOREG

Objectives: Forty to sixty percent of patients with advanced melanoma show primary resistance to PD-1-based immunotherapy, 30-40% of initial responders also progress. Here, we evaluated the outcome of second-line targeted therapy (TT) after progression on PD-1-based immune checkpoint inhibition (ICI) in BRAFV600-mutated melanoma. In addition, we report data on the activity of re-exposure with PD-1-based regimes. Methods: Patients with advanced (non- resectable stage III or IV, AJCC 2017, 8th edition) melanoma progressing on PD-1-based ICI (nivolumab, pembrolizumab or ipilimumab plus nivolumab) and receiving second-line BRAF plus MEK inhibition were identified from the prospective multicenter skin cancer registry ADOREG. Results: We identified 108 patients with unresectable stage III or stage IV melanoma progressing on first-line ICI (nivolumab, pembrolizumab or ipilimumab plus nivolumab) and receiving second-line combined BRAF/MEK inhibition. Seventy- three percent of the cohort presented with primary PD-1 resistant disease. Median progression-free survival ( PFS) on ICI was 2.6 (95% CI 2.2-2.9) months. Median PFS on subsequent TT was 6.6 (95% CI 5.4 -7.8) months. Median OS from start of second-line TT was 16.0 (95% CI 11.2-20.8) months. The 3-year PFS and OS rates on second-line TT were 16% and 30%. The objective response rate (ORR) and disease control rate (DCR) to TT were 42.6% and 55.6%. In patients with brain metastases, the ORR and DCR were 31.4% and 43.1%. Patients without brain metastases showed an ORR and DCR of 52.6% and 66.7%, respectively. Response to first-line ICI was associated with a numerically higher ORR and DCR to second-line TT and improved OS on TT. Twenty-three patients received third-line ICI of whom two patients showed an objective response. Conclusions: BRAF plus MEK inhibition shows meaningful activity and outcome in patients with advanced melanoma resistant to anti-PD-1- based immunotherapy. Rates of long- term benefit and survival in our study were similar to those reported for treatment-naive patients receiving first-line MAPKi

Patient Preferences in Hidradenitis Suppurativa (APProach‐HS): a discrete choice experiment

Summary Background and Objectives Treatment options for moderate‐to‐severe hidradenitis suppurativa (HS) comprise antibiotics, biologics, and different surgical methods. These approaches differ substantially regarding the treatment process, success rates, and adverse events. However, information on patient preferences for HS therapies is hitherto scarce. Our aim was to assess patient preferences for medicamentous and surgical treatment of HS with conjoint analysis. Patients and Methods In this cross‐section study, computerized discrete choice experiments were used to quantify patient preferences for HS therapies decomposed into treatment modality (tablets, subcutaneous injections, surgery with secondary‐intention healing or primary closure), probability of sustained therapeutic success, probability of mild or severe adverse events, and duration of treatment or wound healing. Results Averaged over the cohort (n  =  216 patients with HS), sustained therapeutic success was considered as most important (Relative Importance Score [RIS]: 36.2), followed by the treatment modality (RIS: 24.0), and duration of treatment/wound healing (RIS: 19.9), whereas mild or severe adverse events (RIS: 10.7 or 9.3) were regarded as less relevant. Patients preferred tablets, followed by subcutaneous injections, and disliked surgery with primary closure. Preferences differed significantly dependent on age and affected body regions. Conclusions Awareness of patient preferences is essential for patient‐centered care in HS

Systematic Evaluation of Candidate Blood Markers for Detecting Ovarian Cancer

Epithelial ovarian cancer is a significant cause of mortality both in the United States and worldwide, due largely to the high proportion of cases that present at a late stage, when survival is extremely poor. Early detection of epithelial ovarian cancer, and of the serous subtype in particular, is a promising strategy for saving lives. The low prevalence of ovarian cancer makes the development of an adequately sensitive and specific test based on blood markers very challenging. We evaluated the performance of a set of candidate blood markers and combinations of these markers in detecting serous ovarian cancer.We selected 14 candidate blood markers of serous ovarian cancer for which assays were available to measure their levels in serum or plasma, based on our analysis of global gene expression data and on literature searches. We evaluated the performance of these candidate markers individually and in combination by measuring them in overlapping sets of serum (or plasma) samples from women with clinically detectable ovarian cancer and women without ovarian cancer. Based on sensitivity at high specificity, we determined that 4 of the 14 candidate markers--MUC16, WFDC2, MSLN and MMP7--warrant further evaluation in precious serum specimens collected months to years prior to clinical diagnosis to assess their utility in early detection. We also reported differences in the performance of these candidate blood markers across histological types of epithelial ovarian cancer.By systematically analyzing the performance of candidate blood markers of ovarian cancer in distinguishing women with clinically apparent ovarian cancer from women without ovarian cancer, we identified a set of serum markers with adequate performance to warrant testing for their ability to identify ovarian cancer months to years prior to clinical diagnosis. We argued for the importance of sensitivity at high specificity and of magnitude of difference in marker levels between cases and controls as performance metrics and demonstrated the importance of stratifying analyses by histological type of ovarian cancer. Also, we discussed the limitations of studies (like this one) that use samples obtained from symptomatic women to assess potential utility in detection of disease months to years prior to clinical detection

Are Hox Genes Ancestrally Involved in Axial Patterning? Evidence from the Hydrozoan Clytia hemisphaerica (Cnidaria)

Background: The early evolution and diversification of Hox-related genes in eumetazoans has been the subject of conflicting hypotheses concerning the evolutionary conservation of their role in axial patterning and the pre-bilaterian origin of the Hox and ParaHox clusters. The diversification of Hox/ParaHox genes clearly predates the origin of bilaterians. However, the existence of a "Hox code' predating the cnidarian-bilaterian ancestor and supporting the deep homology of axes is more controversial. This assumption was mainly based on the interpretation of Hox expression data from the sea anemone, but growing evidence from other cnidarian taxa puts into question this hypothesis. Methodology/Principal Findings: Hox, ParaHox and Hox-related genes have been investigated here by phylogenetic analysis and in situ hybridisation in Clytia hemisphaerica, an hydrozoan species with medusa and polyp stages alternating in the life cycle. Our phylogenetic analyses do not support an origin of ParaHox and Hox genes by duplication of an ancestral ProtoHox cluster, and reveal a diversification of the cnidarian HOX9-14 genes into three groups called A, B, C. Among the 7 examined genes, only those belonging to the HOX9-14 and the CDX groups exhibit a restricted expression along the oralaboral axis during development and in the planula larva, while the others are expressed in very specialised areas at the medusa stage. Conclusions/Significance: Cross species comparison reveals a strong variability of gene expression along the oral-aboral axis and during the life cycle among cnidarian lineages. The most parsimonious interpretation is that the Hox code, collinearity and conservative role along the antero-posterior axis are bilaterian innovations

Spezifische Zielgene des Transkriptionsfaktors c-Jun im malignen Melanom

Hintergrund und Ziele: Das Melanom zeigte in den letzten Jahrzehnten eine steigende Inzidenzrate. Durch sein hohes Metastasierungspotenzial gestaltet sich die Therapie des fortgeschrittenen Melanoms weiterhin als schwierig. Die Fehlregulation unterschiedlicher Signalwege und Transkriptionsfaktoren ist an der malignen Entartung von Melanozyten wesentlich beteiligt. Unter den Transkriptionsfaktoren spielt die AP-1-Transkriptionsfaktorfamilie eine wichtige Rolle bei der Pathogenese des Melanoms. Von besonderem Interesse dieser Arbeit war dabei der Transkriptionsfaktor c-Jun. Durch die Regulation spezifischer Zielgene kann c-Jun unterschiedliche biologische Prozesse, wie z. B. Proliferation und Wachstum beeinflussen. Ziel dieser Arbeit war es, spezifische c-Jun-Zielgene im Melanom zu identifizieren und dessen Einfluss auf das Expressionsmuster dieser Zielgene zu analysieren. Methoden: Zur Identifikation möglicher Zielgene des Transkriptionsfaktors c-Jun wurden In-silico-Analysen mittels ENCODE (Encyclopedia of DNA Elements), STRING (Search Tool for the Retrieval of Interacting Genes/Proteins), DAVID (Database for Annotation, Visualization and Integrated Discovery) und MEME (Multiple Em for Motif Elicitation) durchgeführt. Nach Etablierung der Kernextraktion und DNA-Fragmentierung der humanen Melanom-Zelllinie Mel Juso folgte die Chromatin-Immunopräzipitation (ChIP) mit einem c-Jun-Antikörper (sc-1694). Die präzipitierte DNA wurde anschließend mit spezifischen Primer-Pärchen durch eine PCR amplifiziert und die DNA-Amplifikation mittels Agarose-Gelelektrophorese bestätigt. Zur mRNA-Expressionsanalyse der c-Jun-Zielgene wurde eine qRT-PCR mithilfe des LightCycler® 480Systems (Roche Diagnostics, Mannheim) durchgeführt. Hierzu wurden cDNA-Proben unterschiedlicher Melanom-Zelllinien (Mel Juso, A375, Mel Ei, Mel Ju) und normaler humaner Melanozyten (NHEMs) verwendet. Weitere mRNA-Expressionsanalysen wurden mit cDNA-Proben eines c-Jun-Knockdown-Konstruktes (Mel Juso + si c-Jun) und eines Konstruktes mit c-Jun-Überexpression (Hmb2-5 + HAJunMut1) durchgeführt. Ergebnisse und Beobachtungen: Mittels In-silico-Analysen konnten 44 potenzielle Zielgene des Transkriptionsfaktors c-Jun identifiziert, auf bekannte und mögliche Assoziationen zueinander untersucht und in sieben Gruppen unterschiedlicher biologischer Prozesse kategorisiert werden. Für die folgenden Analysen wurde eine Auswahl von sechs Zielgenen getroffen, die unter anderem bereits als tumorrelevante Gene beschrieben wurden: WEE1, PVR, LGALS3, MAP1LC3B, NFATc2 und FosB. In den mittels ENCODE angereichterten Promotor- bzw. Enhancer-Regionen der jeweiligen Zielgene konnte für WEE1, PVR, LGALS3 und FosB eine klassische AP-1-Bindungssequenz (5‘-TGA (C/G) TCA-3‘) detektiert werden. Für MAP1LC3B und NFATc2 wurde eine mögliche AP-1-Bindungssequenz mit jeweils einem unterschiedlichen Basenpaar identifiziert (MAP1LC3B: 5‘-TGA T TCA-3‘; NFATc2: 5‘-TGA C ACA-3‘). Mittels PCR wurde anschließend eine direkte Bindung von c-Jun an die Promotor- bzw. Enhancer-Region der sechs Zielgene bestätigt. Mithilfe von Expressionsanalysen in verschiedenen humanen Melanom-Zelllinien und normalen humanen Melanozyten konnte eine unterschiedliche Expression der sechs Zielgene nachgewiesen werden. Dabei lagen WEE1, PVR, NFATc2 und FosB in den untersuchten humanen Melanom-Zelllinien erhöht exprimiert vor, wohingegen MAP1LC3B und LGALS3 eine verminderte Expression aufwiesen. Abschließend wurde die direkte Regulation der sechs Zielgene durch den Transkriptionsfaktor c-Jun untersucht. Der Knockdown von c-Jun führte zu einer vermehrten Expression von MAP1LC3B und LGALS3 und deutet auf eine negative Regulation durch c-Jun hin. Demgegenüber führte eine Überexpression von c-Jun zu einer vermehrten Expression von WEE1, PVR, NFATc2 und FosB, was auf eine positive Regulation hinweist. Schlussfolgerung: Die Identifikation spezifischer Zielgene des Transkriptionsfaktors c-Jun im Melanom lässt Rückschlüsse auf dessen regulatorische Einflüsse auf unterschiedliche biologische Prozesse ziehen. Die durch c-Jun regulierten Zielgene zeigen unter anderem onkogene Eigenschaften auf. Eine gezielte Inhibition von c-Jun und somit auch dessen regulatorischer Einflüsse auf spezifische Zielgene könnte einen neuen Ansatz in der Therapie des Melanoms darstellen.Background and aims: The incidence of malignant melanoma showed an increase in the past decades. Due to its high risk of metastasis the therapy of advanced melanoma is still challenging. The deregulation of different signalling pathways and transcription factors is crucial for the malignant degeneration of melanocytes. The AP-1 transcription factor family plays a significant role in the pathogenesis of melanoma. Specifically, the AP-1 transcriptions factor c-Jun was of particular interest. Through regulation of specific target genes, c-Jun may influence many biological processes, e.g. proliferation and growth. The aim of this study was to identify specific target genes of c-Jun in melanoma and to analyse its influence on the expression of these target genes. Methods: In-silico-analyses were performed to identify possible target genes of the transcription factor c-Jun, using ENCODE (Encyclopedia of DNA Elements), STRING (Search Tool for the Retrieval of Interacting Genes/Proteins), DAVID (Database for Annotation, Visualization and Integrated Discovery) and MEME (Multiple Em for Motif Elicitation). After establishing the nuclear extraction and DNA fragmentation of the human melanoma cell line Mel Juso, chromatin-immunoprecipitation (ChIP) with a c-Jun antibody (sc-1694) was performed. The precipitated DNA was further amplified via specific primer-pairs and PCR. The successful amplification was validated via agarose gel electrophoresis. By qRT-PCR using the LightCycler® 480 systems (Roche Diagnostics, Mannheim) mRNA-expression analyses of the c-Jun target genes were performed. To this, cDNA samples of different melanoma cells (Mel Juso, A375, Mel Ei, Mel Ju) and normal human melanocytes (NHEMs) were used. Additional mRNA-expression analyses were performed using cDNA samples of a c-Jun knockdown construct (Mel Juso + si c-Jun) and a c-Jun overexpression construct (Hmb2-5 + HAJunMut1). Results and observations: By In-silico-analyses 44 potential target genes of the transcription factor c-Jun were identified, screened for known and possible associations among each other and categorized in seven groups of different biological processes. Six out of 44 target genes, which are known to be cancer-relevant, were selected for further analyses: WEE1, PVR, LGALS3, MAP1LC3B, NFATc2 and FosB. A classical AP-1 binding sequence (5‘-TGA (C/G) TCA-3‘) was identified in the via ENCODE enriched promoter/enhancer region for WEE1, PVR, LGALS3 and FosB. The promoter/enhancer region for MAP1LC3B and NFATc2 contained an AP-1 binding sequence with one base pair changed (MAP1LC3B: 5‘-TGA T TCA-3‘; NFATc2: 5‘-TGA C ACA-3‘). A direct binding of c-Jun to the promoter/enhancer region of the six target genes was confirmed by PCR. A differential expression of the six target genes in various melanoma cell lines and normal human melanocytes was detected performing mRNA-expression analyses. Here, WEE1, PVR, NFATc2 and FosB showed an upregulation whereas MAP1LC3B and LGALS3 showed a downregulation in all analysed melanoma cell lines. Finally, the regulation of the 6 target genes through the transcription factor c-Jun was investigated. An upregulation of MAP1LC3B and LGALS3 was shown after c-Jun knockdown, indicating a negative regulation through c-Jun. In contrast, an overexpression of c-Jun led to an upregulation of WEE1, PVR, NFATc2 and FosB, which might point to a positive regulation through c-Jun. Conclusion: Identifying specific target genes of the transcription factor c-Jun in melanoma helps to understand the regulatory impact on various biological processes. Many of the known c-Jun target genes show oncogene characteristics. Therefore, a targeted inhibition of c-Jun and subsequently its regulatory impact on specific target genes could present a new approach in the therapy of malignant melanoma

Long‑Term Outcomes in BRAF‑Mutated Melanoma Treated with Combined Targeted Therapy or Immune Checkpoint Blockade: Are We Approaching a True Cure?

Approximately 50% of all melanomas harbor an activating BRAF mutation. In patients suffering from an advanced melanoma with such a somatic alteration, combined targeted therapy with a BRAF and MEK inhibitor can be applied to significantly increase the survival probability. Nevertheless, resistance mechanisms, as well as negative predictive biomarkers (elevated lactate dehydrogenase levels, high number of metastatic organ disease sites, brain metastasis), remain a major problem in treating melanoma patients. Recently, a landmark overall survival (OS) rate of 34% after 5 years of combined targeted therapy in treatment-naïve patients was reported. On the other hand, patients harboring a BRAF mutation and receiving first-line immune checkpoint blockade with ipilimumab plus nivolumab showed a 5-year OS rate of 60%. As indicated by these data, long-term survival can be reached in melanoma patients but it remains unclear if this is equivalent to reaching a true cure for metastatic melanoma. In this review, we summarize the recent results for combined targeted therapy and immunotherapy in advanced melanoma harboring an activating BRAF mutation and discuss the impact of baseline characteristics on long-term outcome

Adrenal Insufficiency after Long-term use of Topical Glucocorticoids in Patients with Advanced Cutaneous T-cell Lymphomas: A Case Series

Abstract is missing (Short communication