Melanome zählen zu den zehn häufigsten Tumorentitäten weltweit. Bei frühzeitiger Diagnose ist eine Exzision im Gesunden kurativ. Sobald eine Resektion im Gesunden jedoch nicht mehr möglich ist, sinken die Heilungschancen drastisch. Maligne Melanome sprechen wenig auf konventionelle Tumortherapien wie Radiatio und zytostatische Chemotherapie an. Daher werden neue Therapieoptionen in der Melanomtherapie getestet. Neueste Ansätze beziehen sich auf die Modulation von Immunzellen mittels monoklonaler Antikörper sowie die Modifikation der Signaltransduktion über die Mitogen-aktivierte Protein Kinase Kinase (MAPKK = MEK), BRAF und c-KIT. Auch Ionenkanäle stellen eine vielversprechende, zukünftige Option in der Behandlung maligner Melanome dar. Ich konnte zeigen, dass neben der bereits beschriebenen funktionellen Expression des „transient receptor potential“ Kanals TRPM8 in Melanomzelllinien auch TRPA1 in verschiedenen Melanomzelllinien exprimiert und funktionell ist. Die Phytopharmaka Senföl (Allylisothiozyanat; AITC) und Zimtaldehyd zeigen in Melanom-Modellen antitumoröse Effekte. Zudem sind beide Substanzen potente Stimulatoren von TRPA1. In dieser Arbeit wurde untersucht, ob AITC und Zimtaldehyd TRPA1-vermittelt die Proliferation, Apoptose und Migration von Melanomzellen beeinflussen. Das Vorkommen von TRPA1 in verschiedenen Melanomzelllinien wurde auf molekularbiologischer Ebene, mit fluorometrischen Bestimmungen des TRPA1-vermittelten Ca2+-Einstroms sowie in elektrophysiologischen Messungen nachgewiesen. Anschließend wurde die funktionelle Relevanz von TRPA1 bezüglich tumorhemmender Eigenschaften geprüft. Durch die Anwendung von TRPA1-Blockern konnte die AITC- und Zimtaldehyd-induzierte Verminderung der Proliferation nicht aufgehoben werden. Auch bezüglich der Migration und Apoptose konnte keine Korrelation zu einer TRPA1-Modulation festgestellt werden. Daher scheinen die durch AITC und Zimtaldehyd induzierten Effekte höchstwahrscheinlich nicht durch TRPA1 vermittelt zu werden.:1 Bibliographische Beschreibung 4 2 Abkürzungsverzeichnis 6 3 Einführung 8 3.1 Die Superfamilie der TRP-Kanäle und ihre Expression in malignen Tumoren 8 3.2 Weitere Ionenkanalentitäten im malignen Melanom 10 3.3 Klassifikation und Therapie maligner Melanome 11 3.4 Naturstoffe mit TRP-Kanal-aktivierenden Eigenschaften in der Therapie maligner Tumore 13 3.5 TRPA1 - ein Ionenkanal mit chemosensorischen Eigenschaften 14 4 Ableitung der Fragestellung 17 5 Publikation 18 6 Zusammenfassung der Arbeit 28 7 Literaturverzeichnis 31 8 Anlagen 36 8.1 Erklärung über die eigenständige Abfassung der Arbeit 36 8.2 Curriculum Vitae 37 8.3 Publikationen 39 8.4 Danksagung 40

Le mélanome uvéal (MU) est une tumeur maligne intraoculaire rare qui touche environ 500 français par an. Malgré un traitement efficace de la tumeur primaire, la moitié des patients développent des métastases principalement hépatiques dans les années qui suivent le diagnostic. En dépit des nombreux efforts réalisés, les thérapies systémiques adjuvantes restent peu efficaces sur le MU métastatique. L’identification de gènes pronostiques et/ou causals du développement métastatique pourrait ainsi permettre des avancées considérables dans la compréhension de cette pathologie et le développement de nouvelles thérapies. Notre laboratoire a identifié la surexpression du gène codant la protéine tyrosine phosphatase PRL-3/PTP4A3 comme hautement prédictive du risque métastatique et du devenir des patients atteints de MU. Sa surexpression est également décrite dans de nombreux autres types de cancers humains métastatiques. La surexpression de PRL-3 dans des cellules de MU augmente significativement la migration cellulaire in vitro et l’invasion in vivo de manière dépendant de son activité catalytique, ce qui suggère un rôle direct de PRL-3 dans le processus métastatique du MU. De plus, nous avons montré qu’empêcher l’ancrage membranaire de PRL-3 en utilisant un inhibiteur de farnésylation (FTI-277) abolit la migration induite par PRL-3 dans les cellules de MU, ce qui révèle l’importance de son ancrage membranaire pour la migration cellulaire. Le but de ma thèse a été d’identifier et de caractériser des substrats cellulaires, et plus particulièrement membranaires, de PRL-3 qui seraient impliqués dans le processus métastatique du MU. Mes résultats montrent que la surexpression de PRL-3 dans des cellules de MU, empêchent l’adhérence des cellules au collagène I et la maturation de structures d’adhérence (FAs) en anneaux impliquant l’intégrine β1 (Itg β1), de manière dépendante de son activité catalytique et de son ancrage membranaire. Nous avons également montré que PRL-3 interagit avec l’Itg β1 et la déphosphoryle sur son motif de phosphorylation intracytoplasmique riche en S/T (T788 et T789), dont l’état de phosphorylation est connu pour réguler l’adhérence cellulaire. Ainsi, mes travaux de recherche ont permis d’identifier PRL-3 comme régulateur des structures d’adhérence à la matrice extracellulaire (MEC) au travers de la régulation de l’Itg β1 et potentiellement de la kinase FAK. De plus, dans les FAs nous avons observé que PRL-3 régule spécifiquement l’agrégation de l’Itg β1 mais pas celle de l’Itg β3, ainsi nous émettons l’hypothèse que cette régulation par PRL-3 serait différentielle entre les intégrines et dépendante de la MEC. Dans le MU, la migration accrue des cellules par PRL-3 peut également être expliquée par une accumulation de la métalloprotéase MT1-MMP/MMP14 à la surface des cellules. Cette protéine transmembranaire est responsable de la dégradation de différents substrats de la MEC et peut-être trouvée dans les FAs. Un travail auquel j’ai contribué, a montré que PRL-3 favoriserait l’accumulation de MMP14 à la membrane plasmique par l’accélération de son trafic vésiculaire. Enfin durant la dernière année de ma thèse, nous avons entrepris de tester les effets de la pentamidine, un antiparasitaire aux propriétés anti-cancéreuses, sur l’inhibition de l’activité de PRL-3. In vivo, la pentamidine induirait une inhibition moyenne de la croissance tumorale dans un modèle de xénogreffes murins de MU et de métastases. Les essais in vitro sont encore en cours. / Uveal Melanoma (UM) is a rare tumor that affects around 500 French people each year. Despite a successful treatment of the primary tumor, 50% of patients develop metastasis primarily to the liver in the years following diagnosis. Currently, systemic adjuvant therapy has been unsuccessful for effective treatment. As such, identifying genes involved in both prognosis and metastasis is important for a better understanding of this disease and in turn for designing better treatment strategies. Our group previously identified that overexpression of the gene encoding PRL-3/PTP4A3, a protein tyrosine phosphatase, is highly correlated with metastatic tumor progression and predicts poor prognosis in patients with UM. It is also known that PRL3 is implicated in the metastatic process of various cancers. Overexpression of PRL-3, but not the inactive mutant of PRL3 (C104S), in an ocular melanoma cell line significantly increased cell migration in vitro and invasion in vivo, suggesting a direct role of PRL3 in the metastatic process in UM. We also showed that FTI-277, a farnesyltransferase inhibitor that prevents PRL-3 anchorage to the plasma membrane, abolishes PRL-3-induced UM cell migration on collagen I, suggesting that PRL-3 anchorage is important for cell migration. The aim of my thesis was to identify intracellular, and in particular, membrane substrates that could play a role in UM metastasis. My results show that PRL3 overexpression in UM cells prevents both the spreading of cells to the extracellular matrix (ECM), and the formation of large focal adhesions structures (FA) involving integrin β1 (Itg b1).These biological effects are PRL-3-activity and anchorage dependent. We show that PRL-3 interacts with and dephosphorylates Itg b1 on cytoplasmic threonine 788 and 789, residues that are known to be involved in cell adhesion. Our results identify PRL-3 as a new regulator of cell adhesion structures to the ECM via the regulation of Itg b1 and most likely the focal adhesion kinase (FAK). In FA, we observed that PRL-3 specifically regulates the aggregation of Itg b1 but does not affect integrin β3, so we suppose that this regulation could be specific to certain integrins. In UM cells, the PRL-3-induced cell migration could also be explained by membrane accumulation of the metalloprotease MT1-MMP/MMP14 in the presence of PRL3. This transmembrane proteinase is responsible for ECM degradation and can be found in FA. Moreover, we demonstrated that the vesicular trafficking of MT1-MMP is accelerated in the presence of active PRL-3 but not in presence of the inactive mutant of PRL-3 (C104S). During the last year of my thesis, another aspect of my PhD project was to study the biological effect of pentamidine, an antiparasitic which is known to inhibit the phosphatase activity of PRLs in vitro. In vivo, we show that pentamidine treatment induces a decrease of tumor growth in a UM patient-derived xerograph model. Overall, the results of my thesis suggest that PRL-3 plays an important role in UM metastasis.

L’hypothèse de l’hormono-dépendance du mélanome cutané est en débat dans la littérature depuis de nombreuses années. Dans ce contexte, il est essentiel de mieux connaître l’influence potentielle de la prise de traitements hormonaux sur le risque de ce cancer.L’objectif de ce projet de thèse était d’étudier l’influence de la prise de traitements hormonaux (contraceptifs oraux, traitements de l’infertilité, progestatifs pris seuls avant la ménopause et traitements hormonaux de la ménopause) sur le risque de mélanome cutané chez les femmes.Le projet a été conduit principalement à partir de la cohorte prospective E3N (Étude épidémiologique auprès de femmes de l’Éducation nationale) portant sur environ100 000 femmes âgées de 40 à 65 ans à l’inclusion en 1990. L’étude E3N inclut des données détaillées sur le profil pigmentaire, l’exposition solaire et la prise de traitements hormonaux des participantes, ainsi que sur la survenue éventuelle de mélanome cutané, dont les cas ont été confirmés par histologie (761 cas confirmés entre 1990 et 2011). Des données complémentaires sur l’exposition solaire, disponibles pour une fraction de la cohorte E3N, ont également été utilisées. Ces données détaillées sont issues de l’enquête cas-témoin nichée E3N-SunExp menée en 2008. Les analyses ont ensuite été étendues au consortium de cohortes européennes prospectives EPIC (European Prospective Investigation into Cancer and Nutrition), sur près de 334 500 femmes parmi lesquelles 1 734 cas de mélanomes incidents ont été diagnostiqués.Les résultats mettent en évidence des associations modestes entre la prise de traitements hormonaux et le risque de mélanome et ne soutiennent pas l’hypothèse de l’hormono-dépendance de ce cancer. Cependant, ils suggèrent un comportement d’exposition intentionnelle aux UV chez leurs utilisatrices, ce qui ouvre une nouvelle perspective pour les recherches futures dans ce domaine. / The hypothesis of a hormonal dependence of cutaneous melanoma has been debated in the literature over past decades. Within this context, it is critical to increase our knowledge on the influence of exogenous hormone use on the risk of this cancer.The aim of this project was to explore the influence of hormonal treatments (oral contraceptives, fertility drugs, premenopausal use of progestogens, and menopause hormone therapy) on the risk of cutaneous melanoma in women.The project was mainly based on the large E3N (Étude épidémiologique auprès de femmes de l’Éducation Nationale) prospective cohort, which included about 100,000 women aged 40-65 years at inclusion in 1990. The cohort collected data on pigmentary phenotype, sun exposure, hormonal treatment use, and medical data including cutaneous melanomas, which were ascertained through pathology reports (761 incident cases between 1990 and 2011). Additional information on sun exposure, available for a portion of the E3N cohort, was used. These detailed data were collected as part of the E3N-SunExp case-control study launched in 2008.Analyses were extended to the EPIC consortium (European Prospective Investigation into Cancer and Nutrition), which includes about 334 500 women and 1,734 incident melanoma cases.The results arising from this project show modest associations between exogenous hormone use and melanoma risk and suggest no strong direct influence of hormones on melanoma development. However, results suggest intentional UV exposure behaviors in exogenous hormone users, which opens new perspectives for future investigations in this field.

Le mélanome est le cancer de la peau le plus mortel. Il est caractérisé par une grande hétérogénéité et une reprogrammation métabolique importante qui lui confère l’habileté de promouvoir des programmes immunosuppressifs et de développer une résistance aux traitements. Cette capacité permet au mélanome d’agir sur le microenvironnement tumoral et d’échapper à l’immunosurveillance du système immunitaire. La famille des peroxisome-proliferator activated receptor gamma coactivator 1 (PGC-1s) est un joueur clé du métabolisme cellulaire en régulant la biogenèse mitochondriale, la phosphorylation oxydative et la détoxification du stress oxydatif. Des études ont montré que l’expression de PGC-1α module la fonction mitochondriale. Les fonctions de PGC-1β et PRC, les 2 autres membres de cette famille, dans le mélanome restent largement inexplorées. Ce mémoire montre pour la première fois que l’expression des PGC-1s est non seulement associée à l’expression de plusieurs molécules pro-inflammatoires (IL-8, TNF, IL-1), mais aussi à l’expression de molécules immunosuppressives (CD73, PD-L2, Galectin-9) pouvant contrôler la réponse immunitaire. Par l’utilisation d’inhibiteurs ciblant des voies de signalisation de l’immunité innée, nous avons montré que la régulation de ces molécules s’effectue via MEK et IKK dans les cellules déplétées en PGC-1β. La déplétion en PGC-1 altère la fonction mitochondriale, induisant l’expression de p21 et l’arrêt du cycle cellulaire d’une manière soutenue et via un mécanisme indépendant des dérivés réactifs de l’oxygène (ROS). Nos travaux montrent que les PGC-1s possèdent d’importantes fonctions immunitaires dans le mélanome qui peuvent potentiellement dicter la croissance tumorale, l’évasion cellulaire et la réponse aux thérapies anticancéreuses. / Melanoma is the deadliest form of skin cancer. It is defined by great heterogeneity and extensive metabolic reprogramming which gives it the ability to promote immunosuppressive programs and develop therapy resistance. This ability allows melanoma to define the tumor microenvironment and escape the immunosurveillance of the immune system. The peroxisome-proliferator activated receptor gamma coactivator 1 (PGC-1s) family is a key player in cell metabolism by regulating mitochondrial biogenesis, oxidative phosphorylation and oxidative stress detoxification. Studies have shown that the expression of PGC-1α is linked to increased mitochondrial function and the metastatic potential of melanoma. The functions of PGC-1β and PRC, the other 2 members of this family, in melanoma remain largely unexplored. This thesis shows for the first time that the expression of PGC-1s is not only associated with the expression of several pro-inflammatory molecules (IL-8, TNF, IL-1) but also with the expression of immunosuppressive molecules (CD73, PD-L2, Galectin-9) which can control the immune response. Using inhibitors targeting innate immunity signaling pathways, we have shown that the regulation of these molecules occurs via MEK and IKK in PGC-1β depleted melanoma cells. Depletion of PGC-1 impairs mitochondrial function and leads to p21 induction and cell cycle arrest in a sustained manner by a reactive oxygen species (ROS)-independent mechanism. Our work shows that PGC-1s have important immune functions in melanoma that can potentially dictate tumor growth, cell evasion and response to cancer therapies.

Stem cell therapy may present an effective treatment for metastatic brain cancer and glioblastoma. Here we posit the critical role of a leaky blood-brain barrier (BBB) as a key element for the development of brain metastases, specifically melanoma. By reviewing the immunological and inflammatory responses associated with BBB damage secondary to tumoral activity, we identify the involvement of this pathological process in the growth and formation of metastatic brain cancers. Likewise, we evaluate the hypothesis of regenerating impaired endothelial cells of the BBB and alleviating the damaged neurovascular unit to attenuate brain metastasis, using the endothelial progenitor cell (EPC) phenotype of bone marrow-derived mesenchymal stem cells. Specifically, there is a need to evaluate the efficacy for stem cell therapy to repair disruptions in the BBB and reduce inflammation in the brain, thereby causing attenuation of metastatic brain cancers. To establish the viability of stem cell therapy for the prevention and treatment of metastatic brain tumors, it is crucial to demonstrate BBB repair through augmentation of vasculogenesis and angiogenesis. BBB disruption is strongly linked to metastatic melanoma, worsens neuroinflammation during metastasis, and negatively influences the prognosis of metastatic brain cancer. Using stem cell therapy to interrupt inflammation secondary to this leaky BBB represents a paradigm-shifting approach for brain cancer treatment. In this review article, we critically assess the advantages and disadvantages of using stem cell therapy for brain metastases and glioblastoma. / National Institutes of Health / Revisión por pares

Vitiligo is an autoimmune skin disease in which the pigment producing cells of the epidermis, melanocytes, are targeted for destruction by CD8+ T cells specific for melanocyte/melanoma-shared antigens. Previous work has identified IFNg as the central cytokine driving disease pathogenesis in both human patients and in our mouse model of vitiligo. IFNg signaling induces production of the chemokines CXCL9 and CXCL10, which trigger autoreactive T cell migration into the epidermis where effector T cells can target and destroy melanocytes. However, both IFNg and type I IFN signaling through activation of STAT1 proteins can induce transcription of the chemokines CXCL9 and CXCL10. Therefore, it seems reasonable that type I IFN signaling may also contribute to disease pathogenesis. The role of type I IFN in vitiligo is still unclear. Genome wide association studies identified multiple genes within the type I IFN pathway including TICAM1 and IFIH1 as susceptibility loci in vitiligo. One additional study reported increased epidermal staining of CD123, a marker expressed by pDCs, and the type I IFN induced gene MX1 in vitiligo patient skin. However, this study did not show any functional data to support the role of type I IFN signaling in vitiligo pathogenesis. Since the role of type I IFN in vitiligo is ill-defined, we used two different mouse models of vitiligo to functionally determine the role of type I IFN in disease by inducing vitiligo in hosts which lack the type I IFN receptor (IFNaR). In the first model, we induced vitiligo by adoptive transfer of melanocyte-specific CD8 T cells, which are activated in vivo by infection with recombinant vaccinia virus (VACV) expressing their cognate antigen. Vitiligo induction in IFNaR-deficient mice led to the development of severe disease compared to wild type mice. Acceleration and severity of disease was characterized by increased early recruitment of melanocyte-specific CD8 T cells to the skin, increased production of effector cytokines TNFa and IFNg, and reduced PD-1 expression. Increased production of IFNg by CD8 T cells in the skin of IFNaR-deficient mice led to increased expression of the chemokines CXCL9 and CXCL10 driving disease progression. IFNaR-deficient mice also displayed significantly increased VACV titters compared to wild type hosts. This data reveals a role of type I IFN in the clearance of recombinant VACV. This data also suggests that persistent VACV infection and prolonged antigen exposure in IFNaR deficient hosts is likely driving enhanced activation of melanocyte specific CD8 T cells and the subsequent development of severe vitiligo. Since melanocytes and melanoma cells express shared antigens that can be recognized by CD8 T cells, and because the development of vitiligo after melanoma immunotherapy is a positive prognostic factor for patients, we asked whether VACV vaccine therapy in IFNaR deficient mice would enhance the anti-tumor response to melanoma. B16-F10 inoculated wild type and IFNaR-deficient mice received adoptive transfer of melanocyte-specific CD8 T cells in combination with vaccinia virus expressing their cognate antigen to activate the cells in vivo. Treatment of adoptive T cell transfer and infection with VACV in IFNaR-deficient mice revealed significantly reduced tumor burden compared to wild type mice. Improved tumor regression in IFNaR-deficient hosts was characterized by increased infiltrating cytotoxic T lymphocytes and reduced PD-1 expression. These results further demonstrate that in the absence of type I IFN, hosts mount a robust cytotoxic CD8 T cell response against melanocyte/melanoma antigens and this is likely a result of persistent VACV that leads to prolonged CD8 T cell priming. As a result, IFNaR deficient hosts kill tumor cells more efficiently. To determine whether type I IFN regulates disease pathogenesis in the absence of virus infection, we generated a model of vitiligo in which bone marrow derived dendritic cells (BMDCs) pulsed with the cognate antigen were used to prime melanocyte-specific T cells in place of the viral vector. Induction of vitiligo in IFNaR-deficient hosts using BMDCs revealed no significant differences in disease score compared to wild type hosts. This data clearly demonstrates that type I IFN, in contrast to IFNg, is not required during the effector stage of vitiligo pathogenesis in mice. However, since we intentionally activate transferred melanocyte-specific CD8 T cells with VACV or BMDCs expressing their cognate antigen, our mouse models may circumvent the role of type I IFNs in initiating activation of autoreactive cells and driving autoimmunity. Type I IFN is critical for providing innate immune signals that drive the priming of autoreactive T cells through maturation of DCs by inducing antigen presentation, co-stimulatory molecule expression, and migration to the lymph nodes to encounter naïve T cells. Our mouse models of vitiligo may not capture this process. We have addressed this question by using a TLR ligand to activate BMDCs before transfer into hosts. In fact, activation of BMDCs before transfer leads to significantly enhanced vitiligo in mice and this is partially a result of type I IFN signaling on host cells. Thus, we provide evidence that type I IFNs can enhance the activation of melanocyte-specific CD8 T cells and drive autoimmunity. Collectively, our results show that type I IFN signaling has disparate effects on autoreactive T cell priming in a context dependent manner. We reveal that although type I IFN is not required for the effector phase of vitiligo in mice, maturation of DCs and subsequent type I IFN production can enhance the priming of autoreactive T cells and enhance vitiligo severity. Our studies also reveal that type I IFN is required to clear recombinant attenuated VACV infection and vaccine administration in IFNaR deficient hosts led to a robust autoreactive and anti-tumor response. These insights describing the role of type I IFN in autoimmunity and tumor immunology could have important implications for T cell dependent tumor immunotherapy.

No description available.

The dissertation is focused on the study of intercellular interactions in skin tumors. It is based on 5 original publications that cover several topics. We studied the origin of tumor-associated fibroblasts concerning the primary tumor population. We demonstrated using a mouse model that tumor-associated fibroblasts are produced from the host organism and thus did not arise from transformation directly from tumor cells. We also investigated the relationship between tumor-associated fibroblasts and keratinocytes. We have shown that tumor-associated melanoma fibroblasts affect keratinocytes which, under their influence, acquire the features typically observed in migrating cells and cells undergoing epithelial-mesenchymal transition. We studied the interactions between healthy fibroblasts and tumor cells. We have demonstrated that fibroblasts acquired from healthy skin from a patient suffering from melanoma are significantly different from control fibroblasts of healthy donors in the expression profile. Changes in distal fibroblasts support the view of melanoma as a systemic disease. We have further demonstrated that melanoma-associated fibroblasts do not carry a BRAF mutation, in contrast to BRAF positivity of melanoma cells. And therefore, they did not arise from the transition from melanoma. The...

Die Entwicklung der neuen Systemtherapien führte in den letzten Jahren zu einer bedeutenden Verbesserung des Langzeitüberlebens bei Patienten mit metastasiertem Melanom. Die verschiedenen Therapieoptionen mit ihren entsprechenden jeweiligen Toxizitäten sowie die Unterschiede im Therapieansprechen auf die differenten Therapieformen machen es erforderlich, anhand von prädiktiven Faktoren und Biomarkern Therapieentscheidungen zu treffen, um individuell die beste Therapie für den Patienten auszuwählen. Die Tumorwachstumskinetik wird dabei immer wieder als relevanter Faktor genannt. Ihr genauer Einfluss sowie die genaue Definition einer schnellen Tumorkinetik ist jedoch bis heute nicht klar definiert. Diese Arbeit soll dazu beitragen, die prognostische Wertigkeit der Metastasenwachstumsrate (MGR) als prädiktiven Marker vor dem Start einer Systemtherapie zu untersuchen. Konkret soll die Hypothese geprüft werden, ob eine schnelle MGR vor Therapiebeginn einen negativen Einfluss auf das Therapieansprechen sowie das klinische Outcome hat. Vor diesem Hintergrund wurde retrospektiv ein Patientenkollektiv von insgesamt 115 Patienten, aufgeteilt in drei Kohorten (Monotherapie mit PD-1-Antikörpern: n = 33, Kombinationstherapie mit PD-1- und CTLA-4-Antikörpern: n = 34, zielgerichtete Therapie mit BRAF- +/- MEK-Inhibitoren: n = 48), untersucht. Die Erhebung der Dresdner Daten war dabei Teil eines Verbundprojekts mit 12 teilnehmenden Kliniken, sodass zusätzlich zur Dresdner Auswertung für die Monotherapie aus PD-1-Antikörpern auch eine multizentrische Gesamtanalyse der Daten stattfinden konnte. Für die Datenerhebung wurden alle Patienten in Betracht gezogen, die seit Zulassung der einzelnen Therapieformen aufgrund eines kutanen, uvealen, mucosalen oder okkulten Melanoms im Stadium IIIB oder höher am Universitätsklinikum Dresden mit einer entsprechenden Systemtherapie behandelt wurden und eine messbare Metastasierung (CT/MRT/PET-CT) im Baseline-Staging sowie einem Pre-Baseline-Staging aufwiesen. Bei fehlenden Follow-Up-Daten sowie einer adjuvanten Therapiesituation wurden die Patienten aus der Datenerhebung ausgeschlossen. Die MGR wurde für den Zeitraum zwischen Pre-Baseline-Staging und dem Baseline-Staging bestimmt. Mittels Kaplan-Meier-Überlebenskurven sowie univariater und multivariater Cox-Regression wurde der prognostische Einfluss der MGR analysiert. Die Untersuchung des Einflusses der MGR auf das Therapieansprechen erfolgte mithilfe des Fisher-Tests und des Mann-Whitney-U-Tests. In der Dresdner Studie war eine hohe MGR (>3,9 mm/Monat) mit einem signifikant schlechteren progressionsfreien Überleben (PFS) für Patienten unter einer zielgerichteten Therapie verbunden. Zusätzlich zeigten diese Patienten ein signifikant schlechteres Ansprechen auf die Therapie mit BRAF- und MEK-Inhibitoren, im Vergleich zu Patienten, die eine MGR < 3,9 mm/Monat aufwiesen. Für das Gesamtüberleben (OS) zeigte sich trotz geringer Fallzahl sowohl in der Kohorte der zielgerichteten Therapie als auch in der Kohorte der kombinierten Immuntherapie mit Ipilimumab und Nivolumab ein Trend zu einem kürzeren OS, wenn eine hohe MGR vorlag. Dieser Trend konnte für die kombinierte Immuntherapie zusätzlich auch für das PFS festgestellt werden. Für die PD-1-Antikörper-Monotherapie konnte anhand der Dresdner Daten kein signifikanter Unterschied für das OS und PFS zwischen Patienten mit niedriger und hoher MGR festgestellt werden. In der Gesamtbetrachtung stellt die MGR einen prognostischen Marker für Patienten unter einer zielgerichteten Therapie mit BRAF- und MEK-Inhibitoren sowie in Zusammenschau mit den Ergebnissen der Gesamtstudie auch für Patienten unter einer PD-1-Antikörper-Monotherapie dar. Zusätzlich ergaben sich innerhalb dieser Arbeit Hinweise darauf, dass dies auch für die Immunkombinationstherapie mit Ipilimumab und Nivolumab zutrifft. Um die prognostische Wertigkeit der MGR für die unterschiedlichen Systemtherapien gleichermaßen beurteilen zu können, sind weitere Untersuchungen an größeren Patientenkollektiven notwendig. / The development of new system therapies has resulted in an important improvement in the long term survival of patients with a metastatic melanoma. Different therapy options with their respective individual toxicity, as well as differences in the reactions to different forms of therapy, require therapeutic decisions on the basis of predictive factors and bio markers in order to choose individually the best therapy for the patient. Tumor growth kinetics is often considered to be a relevant factor. Its specific influence as well as the exact definition of fast tumor kinetics has, however, not yet been clearly defined. This paper is supposed to support the examination of the prognostic value of the metastatic growth rate (MGR) as a predictive marker before a system therapy is introduced. The hypothesis that a fast MGR before a therapy begins has a negative influence on the therapy response and the clinical outcome has to be examined. On the basis of this hypothesis a patient collective of 115 patients in total, divided into three cohorts (mono therapy with PD-1-antibodies: n=33, combination therapy of PD-1- and CTLA-4-antibodies: n=34, target-orientated therapy with BRAF- t/- MEK inhibitors: n=48) was examined retrospectively. The collection of the Dresden data was part of a cooperative project of 12 participating clinics resulting in a multi-centric overall analysis of the data in addition to the Dresden evaluation of the mono therapy of PD1-antibodies. For the collection of the data those patients were considered who had been treated at Dresden university hospital since the approval of their individual form of therapy with a respective system therapy because of a cutaneous, uveal, mucosal or occult melanoma stage IIIB or higher and who showed a measurable metastasis (CT/MRT/PET-CT) in baseline staging as well as in pre-baseline staging. Patients without any follow-up data and an adjuvant therapy were excluded from the collection of data. The MGR was fixed for the timespan between pre-baseline staging and baseline staging. The prognostic influence of the MGR was analysed by means of the Kaplan-Meier-survival curve as well as univariate and multivariate cox regression. The study of the influence of MGR on the therapy response was based on the Fisher-Tests and the Mann-Whitney-U-Tests. The Dresden study showed the correlation between a high MGR (>3,9 mm/month) and a significantly worse progression free survival (PFS) for patients having a target-orientated therapy. Additionally, those patients showed a significantly weaker reaction to the therapy with BRAF - and MEK-inhibitors in comparison to patients who had an MGR < 3,9mm/month. Considering the overall survival (OS) of those with a fast MGR a trend to a shorter OS could be established, in spite of low case numbers in the cohort of the target-orientated therapy as well as in the cohort of the combined immune therapy with Ipilimumab and Nivolumab. This trend could be found for the combined immune therapy as well as for the PFS. Referring to the anti-PD1- mono therapy no significant difference could be found for the OS and the PFS between patients with faster and slower MGR. In the overall view the MGR proves to be a prognostic marker for patients having a target-orientated therapy with BRAF- and MEK-inhibitors and, also looking at the results of the overall study, for patients having a PD-1- antibody mono therapy. Additionally, this study revealed a significant indication that this also applies to the immune combination therapy of Ipilimumab and Nivolumab. Further studies with larger collectives of patients are necessary to be able to likewise judge the different prognostic valency of the MGR for the different system therapies.

Melanoma is a life threatening type of skin cancer with numerous fatal incidences all over the world. The 5-year survival rate is very high for cases that are diagnosed in early stage. So, early detection of melanoma is of vital importance. Except for several techniques that clinicians apply so as to improve the reliability of detecting melanoma, many automated algorithms and mobile applications have been developed for the same purpose.In this paper, deep learning model designed from scratch as well as the pretrained models Inception v3 and VGG-16 are used with the aim of developing a reliable tool that can be used for melanoma detection by clinicians and individual users. Dermatologists who use dermoscopes can take advantage of the algorithms trained on dermoscopical images and acquire a confirmation about their diagnosis. On the other hand, the models trained on clinical images can be used on mobile applications, since a cell phone camera takes images similar to them.The results using Inception v3 model for dermoscopical images achieved accuracy 91.4%, sensitivity 87.8% and specificity 92.3%. For clinical images, the VGG-16 model achieved accuracy 86.3%, sensitivity 84.5% and specificity 88.8%. The results are compared to those of clinicians, which shows that the algorithms can be used reliably for the detection of melanoma.