Angiogenesis, the formation of new blood vessels from pre-existing ones, is a crucial step in many pathologies, including tumor growth and metastasis. We developed an adenovirus vector allowing the 16K hPRL expression, an antiangiogenic factor, in situ. We show 16K hPRL inhibits oxygen-induced retinopathy in mice. Then, using the 16K-Ad vector, we investigated the ability of 16K hPRL to prevent metastatic spread through inhibition of angiogenesis. We show that 16K hPRL administered via adenovirus-mediated gene transfer inhibits tumor growth in a subcutaneous B16-F10 mouse melanoma model by reducing the size and width of tumor vessels. We also show, for the first time, that 16K hPRL considerably reduces the establishment of B16-F10 metastases in an experimental lung metastasis model. These results highlight a potential role for 16K hPRL in anticancer therapy for both primary tumors and metastases. In parallel, we have sought to identify in human 16K PRL (16K hPRL) a peptide that might be responsible for its antiangiogenic activity. Although the 16K fragments of the other three human PRL/GH-family members are also potently antiangiogenic, the sequence similarity of these fragments is low (around 35% similarity between all mammalian PRL/GH sequences). This led us to seek a peculiar common structural feature rather than a similar sequence. We demonstrate that all these fragments possess a 14-amino-acid sequence having the characteristics of a tilted peptide. We show for the first time that tilted peptides exert antiangiogenic activity. The tilted peptides of hPRL and hGH induce endothelial cell apoptosis, inhibit endothelial cell proliferation, and inhibit capillary formation both in vitro and in vivo. These antiangiogenic effects are abolished when the peptides hydrophobicity gradient is altered by mutation. We further demonstrate for the first time that the well-known tilted peptides of SIV gp32 and Alzheimers beta-amyloid peptide are also angiogenesis inhibitors. Taken together these results point to a potential new role for tilted peptides in regulating angiogenesis. Langiogenèse, la formation de nouveaux vaisseaux à partir de vaisseaux préexistants, joue un rôle important dans de nombreuses pathologies incluant la croissance tumorale et la dissémination des métastases. Nous avons développé un vecteur adénoviral (16K-Ad) nous permettant de produire le facteur antiangiogène hPRL 16K directement in situ. Une première analyse permettant détudier les effets de la hPRL 16K produite par lapproche adénovirale a été réalisée in vivo. Nous montrons tout dabord que la hPRL 16K produite est capable de prévenir la néovascularisation rétinienne dans un modèle murin de rétinopathie. Par la suite, en utilisant le vecteur 16K-Ad, nous montrons que la hPRL 16K peut inhiber le développement tumoral dans un modèle murin développant des tumeurs formées par les cellules de mélanome B16-F10 dans le tissu sous-cutané. Cette inhibition de la croissance tumorale est corrélée avec une diminution de la taille des vaisseaux. Nous montrons aussi, pour la première fois, que la hPRL 16K peut considérablement réduire létablissement des métastases B16-F10 dans un modèle expérimental de métastases se développant dans le poumon. Ces résultats soulignent le rôle potentiel de la hPRL 16K dans la thérapie anticancéreuse dirigée contre les tumeurs primaires et les métastases. Parallèlement à ces travaux, nous nous sommes attachés à identifier une région responsable de lactivité de la hPRL 16K. Partant du fait que les fragments 16K des trois autres membres de la famille humaine PRL/GH sont aussi de puissants facteurs antiangiogènes, malgré que leur similarité de séquence soit faible (environ 35% de similarité entre les séquences de la PRL et de la GH), nous avons recherché une caractéristique structurale commune partagée par ces différents fragments. Nous avons identifié un domaine susceptible d'adopter une structure en peptide oblique dans les séquences protéiques des fragments de 16 kDa de la famille PRL/GH. Leffet antiangiogène des peptides obliques de la hPRL 16K et de la hGH 16K est montré dans des expériences menées in vitro et in vivo. Nous montrons également leffet antiangiogène des peptides obliques du peptide de fusion gp 32 du virus SIV et du peptide b-amyloïde.

Initialement, la vision restreinte des MMPs en tant que de simples « destructeurs » de la matrice extracellulaire avait largement sous-estimé limportance et la diversité de leurs contributions aux différentes étapes de la progression tumorale et de la dissémination métastatique. Ces dernières années, les fonctions complexes des MMPs dans la progression tumorale ont été renforcées par la découverte de nombreux nouveaux substrats dégradés, clivés ou activés par les MMPs, incluant non seulement des composants de la matrice extracellulaire mais aussi des récepteurs, des molécules dadhésion et de nombreuses molécules potentiellement bioactives. La complexité et la diversité de laction des MMPs dans la progression tumorale fournissent certaines explications quant à « léchec thérapeutique » de linhibition des MMPs dans les études cliniques de cancers humains. Une compréhension complète et spécifique de la biologie de chaque MMP est néanmoins requise avant de les considérer à nouveau comme des cibles thérapeutiques contre le cancer ou dautres pathologies. Les travaux réalisés dans le cadre de ce doctorat ont contribué à la compréhension des mécanismes moléculaires impliquant la MT1-MMP dans la croissance et langiogenèse tumorale et ont identifié, pour la première fois, une implication de la MT4-MMP dans la croissance tumorale et la dissémination métastatique du cancer du sein. Malgré sa faible activité de dégradation de la matrice extracellulaire, cette protéase à lien glycosylphosphatidyl inositol pourrait promouvoir la progression tumorale en affectant la structure des vaisseaux sanguins intratumoraux.

München, Techn. Univ., Diss., 2004.

Made available in DSpace on 2015-01-26T13:21:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-05-16Bitstream added on 2015-01-26T13:30:37Z : No. of bitstreams: 1 000793042_20160220.pdf: 95696 bytes, checksum: 6438b62215ddf19c908b92f29a7da6a3 (MD5) Bitstreams deleted on 2016-02-22T11:12:45Z: 000793042_20160220.pdf,. Added 1 bitstream(s) on 2016-02-22T11:13:40Z : No. of bitstreams: 1 000793042.pdf: 404315 bytes, checksum: 6fac64781d4b9c7be4daf0ea672a3846 (MD5) / A avaliação microscópica dos linfonodos em cadelas portadoras de carcinoma de mama pode desempenhar um papel fundamental no direcionamento terapêutico, já que a drenagem linfática representa a via mais comum de metástases. Nos últimos anos tem-se desenvolvido técnicas que oferecem excelente acurácia na remoção dos linfonodos, como a Técnica do Linfonodo Sentinela (TLS), que se baseia no princípio de progressão escalonada na drenagem linfática de neoplasias e que atualmente é o principal fator prognóstico e diagnóstico em mulheres portadoras de carcinomas de mama. O objetivo da pesquisa foi avaliar de forma inédita a acurácia da biópsia do linfonodo sentinela em cadelas portadoras de carcinoma de mama. Foram utilizadas 41 cadelas com diagnóstico citológico do tumor. Após injeção do corante azul patente V, procedeu-se a mastectomia, juntamente com a linfadenectomia (sentinelas e não sentinelas). O material foi submetido a cortes seriados e avaliado microscopicamente pela coloração rotineira de H&E e imuno-histoquímica com os marcadores para pan-citoqueratina AE1/AE3 e células mioepiteliais CK14, que revelou que 46,3% (19/41) dos animais apresentavam metástase nodal. Os valores de sensibilidade e especificidade encontrados para a TLS foram de 89,5% e 100%, respectivamente, com índice kappa igual a 0,90 e p<0,0001. Portanto, a TLS oferece alta sensibilidade no diagnóstico precoce de micrometástases, permitindo maior precisão no estadiamento do paciente oncológico e consequentemente, maiores chances de sobrevida / Microscopic evaluation of lymph nodes in bitches with mammary carcinomas may have fundamental importance in guidance the therapy since lymphatic drainage constitutes the most common route for metastasis. In recent years, several techniques have been used to detect and remove Sentinel Lymph Nades (SLN) based on the principIe of stepwise progression of cancer and lymphatic drainage. In human medicine, SLN biopsy and analysis constitutes the main prognostic factor in women with mammary , carcinomas. This study was designed to evaluate, in an unprecedented manner, the accuracy of sentinel lymph no de biopsy in bitches with mammary carcinomas. Forthyone dogs with no predilection for breed or age, with previous cytological diagnosis of mammary carcinoma were submitted to inoculation of patent blue V dye in peritumoral skin, followed by radical mastectomy and radicallymphadenectomy. Lymphanedectomy included sentinel and non- sentinel lymph nades in order to detect lymph nade metastasis. Both groups were compared in order to determine the accuracy of SLN technique in the detection of node metastasis. The lymph nades were subject to a standard serial sectioning method, stained by routine H&E or submitted to immunohistochemistry using pan-cytokeratins AEl/AE3 and the myoepitelial marker CK14. Nineteen animaIs (46.3%) had nodal metastasis. Sensibility and specificity for SLN technique were 89.5 % and 100 %, respectively, with a kappa value of 0.90 (p<O.OOOl). Therefore, SLN biopsy offers high sensitivity in the early diagnosis of lymph no de metastasis allowing a more accurate clinical staging. These results probably impacts on greater chances of patient survival

Made available in DSpace on 2016-08-12T18:48:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-11-26. Added 1 bitstream(s) on 2016-08-12T18:51:08Z : No. of bitstreams: 1 000864495.pdf: 561808 bytes, checksum: dff5ee77e1a3a3047af80805ca02be32 (MD5) / Introdução: O câncer de mama metastático apresenta sobrevida inferior a 25% em cinco anos. A superexpressão do receptor HER2 nas células tumorais destas pacientes piora o prognóstico e diminui ainda mais sobrevida destas pacientes. O lapatinibe é uma droga que bloqueia a porção intracelular do receptor HER2 e esta ação poderia diminuir a proliferação das células neoplásicas diminuindo a progressão do tumor. Foi realizada uma revisão sistemática e metanálise dos estudos que avaliaram a eficácia e segurança do lapatinibe associado à quimioterapia ou terapia hormonal em mulheres com câncer de mama avançado com superexpressõ do HER2. Métodos: A busca dos estudos foi realizada nas bases MEDLINE, EMBASE, WEB OF SCIENCE, CENTRAL, ICTRP, ClinicalTrials.gov, Cochrane Breast Cancer Group. Foram analisados, como desfecho primário, a sobrevida global (SG), a sobrevida livre de progressão (SLP) e a qualidade de vida, e como desfechos secundários os efeitos colaterais relacionados ao tratamento. Resultados: Foram identificados e avaliados 1298 estudos. No final, aplicando os critérios de inclusão, foram selecionados cinco estudos. Juntos, estes estudos totalizaram 1127 pacientes com câncer de mama avançado (TNM estádio III ou IV) com superexpressão de HER2. Em ralação à SG foi observado que Lapatinibe associado a quimioterapia ou terapia hormonal diminui a probabilidade de morte em 20% (Hazard Ratio 0.80, 95% CI 0.69 to 0.94;) p=0,005. Em relação à SLP o Lapatinibe diminui a probabilidade de eventos de progressão tumoral ou morte por qualquer causa em 48% (Hazard Ratio 0.58, 95% CI 0.50 to 0.67 p<0,0001. Nenhum estudo analisou a qualidade de vida. Referente aos efeitos colaterais, os pacientes que receberam Lapatinibe tiveram mais diarreia (RR 2,5, 95 IC 1,82 a 3,43; I2 =86 p <0,00001, aumento de risco de eventos cardíacos (RR 1,74, 95 % IC 1,02 a 2,99; I2 =0% p 0,04), e maior risco de sofrer descontinuidade de... / Introduction: The five years survival of women with metastatic breast cancer accounts less than 25%. HER2 is an epidermal growth factor; the overexpression of HER2 in the cancer cells decreases survival of these patients Lapatinib is a drug that blocks the intracellular domain of HER2. This blocking could stop or diminish the cell proliferation and improve the survival of these patients. WE performed systematic review and metaanalysis of studies that analyzed the efficacy and safety of lapatinib associated with chemotherapy or hormone therapy for patients with advanced breast cancer that overexpress HER2. Methods: A search of the studies was conducted in MEDLINE, EMBASE, WEB OF SCIENCE, CENTRAL, ICTRP, ClinicalTrials.gov, Cochrane Breast Cancer Group. There were analyzed as the primary endpoint, overall survival (OS), progression-free survival (PFS) and the quality of life, and as secondary endpoints side effects, Results: We identified and assessed 1298 studies. Appling the inclusion criteria were selected five studies. Together these studies included a total of 1127 patients with advanced breast cancer (TNM stage III or IV) with overexpression of HER2. In conclusion it was observed that: for OS lapatinib combined with other drugs decreases the probability of death in 20% (Hazard Ratio 0.80, 95% CI 0.69 to 0.94) p = 0.005. For PFS, lapatinib decreases the probability of tumor progression events or death from any cause in 48% (hazard ratio 0:58, 95% CI 0.67 to 0:50 P < 0,0001. None of the studies analyzed the quality of life. Refers to side effects, patients receiving Lapatinib had more diarrhea (RR 2.5, 95 CI 1.82 to 3.43; P<0.00001; increased risk of cardiac events (RR 1.74, 95% CI 1.02 the 2.99; P<0.04), and higher discontinuation of treatment (RR 4.00, 95% CI 1.44 the 11.17). Conclusion: The use of Lapatinib associated with other drugs decreases the probability of death and increases the likelihood of disease progression free ...

Les métastases sont à l'origine dans 90% des cas du décès des patients et sont donc un enjeu thérapeutique majeur. Les traitements anti-angiogéniques qui inhibent la vascularisation tumorale et augmentent l'hypoxie ont une efficacité limitée en clinique. Des liens ont été établis entre l'hypoxie et le changement métabolique des cellules tumorales, l'augmentation des cellules souches cancéreuses, la transition épithélio-mésenchymateuse et la résistance aux traitements mais également la dissémination métastatique. Ces observations nous ont poussés à réévaluer le rôle de l'hypoxie dans la dissémination métastatique du cancer du sein, en nous focalisant sur l'étude du premier inhibiteur endogène de l'angiogenèse identifié, la Thrombospondine 1 (TSP1). En clinique, nous avons observé que l'expression de la TSP1 est corrélée aux marqueurs d'agressivité et qu'elle augmente avec le grade tumoral. Dans un modèle orthotopique de tumeurs mammaires métastatiques, nous avons montré que l'inhibition de la TSP1 réduit fortement l'EMT, l'invasion et les métastases pulmonaires alors même que la vascularisation de la tumeur primaire est densifiée et plus fonctionnelle, réduisant l'hypoxie. Pour déterminer le rôle de la TSP1 dans la survenue des métastases, nous avons effectué des mutations ponctuelles de ce gène dans le domaine d'activation du TGFbéta ou dans celui de la liaison au CD36 qui médie majoritairement les propriétés anti-angiogéniques de la TSP1. La mutation de l'une ou l'autre de ces séquences entraîne une diminution drastique des métastases pulmonaires. Ce travail nous conduit à proposer un nouveau concept thérapeutique, visant à normaliser la vascularisation tumorale et à réduire l'hypoxie par une stratégie "anti-antiangiogénique" basée sur l'inhibition de la TSP1. Cette stratégie devrait inhiber la dissémination métastatique et favoriser la pénétration des traitements chimiothérapeutiques et augmenter l'efficacité de la radiothérapie, dépendante de la présence d'oxygène. / Metastasis are responsible for 90% of cases of patients death and so represent a major therapeutic challenge in oncology. Anti-angiogenic treatments, which inhibit tumor vascularization and increase hypoxia, have limited clinical efficiency. This inefficiency could be a consequence of hypoxia. Indeed, links have been established between hypoxia and: metabolic changes in tumor cells, increase in cancer stem cells, epithelial-mesenchymal transition and resistance to treatment. Experimental and clinical evidence also suggest that hypoxia may directly induce metastatic dissemination. These observations led us to reassess the role of hypoxia in metastatic dissemination by focusing on the study of the first identified endogenous angiogenic inhibitor, the Thrombospondin 1 (TSP1). TSP1 is also an activator of latent TGFbéta. The aim of this work was to study the role of TSP1 in tumor progression and dissemination of metastatic breast cancer. In clinic, we observed that the expression of TSP1 is correlated with markers of aggressiveness and that it increases with tumor grade. In an orthotopic model of metastatic breast tumors, we have shown that inhibition of TSP1 strongly reduce EMT, invasion and lung metastasis meanwhile the vascularization of the primary tumor is densified and more functional, thus reducing hypoxia. To determine the role of TSP1 in the occurrence of metastasis, we generate point mutations of this gene in TGFbéta activation domain or in the binding domain to CD36 that predominantly mediates the anti-angiogenic properties of TSP1. Mutations of either of these sequences result in a drastic decrease in lung metastasis, which confirms the importance of TGFbéta in metastatic dissemination. Our results also demonstrate a direct link between hypoxia and metastasis: the inactivation of anti-angiogenic properties of TSP1, which increases tumor perfusion and decreases hypoxia without affecting primary tumor growth, blocks metastatic dissemination even though the cells have the ability to migrate. This work led us to propose a new therapeutic concept aiming to normalize tumor vascularization and reduce hypoxia by an "anti-antiangiogenic" strategy based on TSP1 inhibition. This strategy should inhibit metastatic spreading, promote the diffusion of chemotherapeutic treatments and increase the efficiency of radiotherapy, dependent on oxygen presence.

Die Arbeit umfaßt erstens einen Literaturüberblick über die charakteristischen Merkmale und Besonderheiten des verrukösen Karzinoms (VK) und die Prognose bei unterschiedlichen therapeutischen Herangehensweisen. Von besonderem Interesse ist die LK-Metastasierung von VK. In der Literaturauswertung konnten dabei 18 Fälle ermittelt werden; dabei waren nur in einem Fall Fernmetastasen vorhanden. Die Prognose von VK ist gut bei einer adäquaten chirurgischen Therapie; die Mitbehandlung des Lymphabstromgebietes sollte nicht routinemäßig erfolgen. Zweiter Schwerpunkt der Arbeit ist eine Auswertung des Patientengutes der Klinik und Poliklinik für Mund-, Kiefer- und Gesichtschirurgie der Charité (24 Patienten) und der Hals-Nasen-Ohrenklinik der Charité (13 Patienten) unter der besonderen Fragestellung des Metastasierungsverhaltens und der Prognose von VK. In 2 Fällen wurden LK-Metastasen eines VK festgestellt. In keinem Fall ist das VK als Todesursache eines Patienten bekannt. Bei 8 Patienten traten Rezidive auf. Es wurde festgestellt, daß größere Tumoren schneller und häufiger rezidivieren. Bei der Hälfte der Patienten trat das Rezidiv 4 bis 9 Monate post operationem auf. Die Früherkennung von VK ist wichtig, denn die Prognose ist umso besser, je kleiner der Tumor ist. / The paper comprises first an assessment of literature about the characteristics, and the particularities of verrucous carcinoma (VC), and prognosis dependent on the different therapies. Of particular interest is the lymph node metastatic invasion of VC. The assessment of literature showed in 18 cases lymph node metastases; one VC of these had distant metastases. The prognosis of VC is excellent with adequate surgical therapy. Lymph node treatment should not be routine. Second point of the paper is the assessment of data of 24 patients with VC of the Clinic for Oral and Maxillo-Facial surgery of the Charité and 13 patients with VC of the Clinic for Otorhinolaryngology of the Charité with special attention on metastatic invasion and prognosis of VC. Two patients had lymph node metastases caused by a VC. In no case VC is known as cause of death. Eight patients had relapses. Half of these patients had relapse 4 to 9 monthes post operationem. It was established that larger tumors relapse more often and faster. Early detection of VC is necessary, because the prognosis is the better the smaller the tumor is.

Canine Mammatumoren (CMT) sind eine der häufigsten Todesursachen bei Hündinnen (SIMON et al. 2001). Durch ihre periphere Lage sind zumeist nicht die Primärtumoren, sondern ihre Metastasen lebensbedrohlich, da diese häufig wichtige Organe wie Lunge Herz und Gehirn befallen (CLEMENTE et al. 2010). Aktuell angewandte Untersuchungsmethoden zur Erkennung von Metastasierung können entweder das invasive Wachstum beschreiben oder bereits bestehende Metastasen detektieren, jedoch fehlen bislang molekulare Marker, die frühzeitig und zuverlässig das metastatische Potential eines CMT anzeigen bevor Metastasen aufgetreten sind. Das Peptidhormon Relaxin ist in zahlreichen physiologischen und pathologischen Situationen beim Menschen als ein Induktor von Matrixmetalloproteinasen (MMP) bekannt (TOO et al. 1984; UNEMORI und AMENTO 1990; PALEJWALA et al. 2001; BINDER et al. 2002; KLONISCH et al. 2007; HENNEMAN et al. 2008). MMP sind Schlüsselenzyme des Bindegewebsabbaus, der jeder Metastasierung vorausgehen muss (WOODHOUSE et al. 1997). Bei der Frau ist im Blut messbares Relaxin ein Marker für metastatischen Brustkrebs (BINDER et al. 2004), für die Hündin scheint dagegen der Relaxinblutspiegel nicht aussagekräftig zu sein (SCHWEIZER 2010). Möglicherweise wird aber Relaxin lokal im caninen Mammagewebe exprimiert wie von GOLDSMITH et al. (1994) und SILVERTOWN et al. (2003) postuliert, so dass es wie in humanen Tumoren auto- oder parakrin Invasivität und Metastasierung (KLONISCH et al. 2007) fördern könnte. Daher sollte in der vorliegenden Arbeit die intratumorale Expression des Relaxins und seiner Rezeptoren sowie bekannter Faktoren des Bindegewebsabbaus untersucht und auf ihre prognostische Eignung überprüft werden. In zwei Studienabschnitten (LAMP et al. 2009; LAMP et al. 2011) wurden CMT-Gewebeproben von n=31 (LAMP et al. 2009) respektive n=59 Hündinnen (LAMP et al. 2011) mittels quantitativer Reverse-Transkriptase-Polymerasekettenreaktion (qRT-PCR) auf ihre Expression von Relaxin, seinen Rezeptoren, RXFP1 und RXFP2, sowie den Matrixmetalloproteinasen MMP-1, -2, -3, -9 und MMP-13, den Östradiolrezeptoren, ERα und ERβ, und dem Progesteronrezeptor (PR) analysiert. In beiden Studienabschnitten wurden die Plasmakonzentrationen der Hormone Relaxin, Östradiol und Progesteron auf mögliche Zusammenhänge mit der lokalen Genexpression überprüft. Im zweiten Studienabschnitt (LAMP et al. 2011) wurde darüber hinaus die Expression von Relaxin und RXFP1 auch immunhistologisch an n=9 CMT-Proben untersucht und in einer Multivarianzanalyse die prognostische Eignung aller untersuchten Parameter getestet. Die Expressionsanalyse konnte zeigen, dass CMT sowohl ein bisher unbekannter Ort der Relaxinexpression beim Hund sind als auch den Hauptrezeptor des Relaxins, RXFP1, exprimieren. Diese Ergebnisse der mRNA-Untersuchung ließen sich immunhistologisch bestätigen. Darüber hinaus ergab die immunhistologische Untersuchung, dass Relaxin vorwiegend im myoepithelialen Anteil der untersuchten CMT exprimiert wird. In den epithelialen CMT-Zellen fand sich die stärkste RXFP1-Reaktivität, so dass RXFP1 mit der von anderen Autoren beschriebenen MMP-2- und MMP-9-Expression in epithelialen Zellen kolokalisiert ist (PAPPARELLA et al. 1997; HIRAYAMA et al. 2002; PAPPARELLA et al. 2002). Die quantitativen Expressionsanalysen zeigten Korrelationen der Expressionsintensitäten von Relaxin, RXFP1 und MMP-2 auf. Die RXFP1 Expression war in dieser Studie sogar ein unabhängiger Marker für Metastasierung mit einem 15-fach höheren Risiko für Metastasierung für Patienten mit einer Expression oberhalb des studienspezifischen Cut-Offs. Alle untersuchten lokalen Genexpressionen waren von den systemischen Plasmakonzentrationen von Relaxin, Östradiol und Progesteron unabhängig. Die Resultate legen eine Bedeutung des intratumoral exprimierten Relaxins für eine auto- oder parakrine Steuerung der MMP-Expression, die für Invasivität und Metastasierung wichtig ist, nahe. Aufgrund des mRNA-Nachweises, der Kolokalisation der Proteine von RXFP1 und MMP-2 und -9 sowie der Korrelation der Genexpression von Ligand (Relaxin), Rezeptor (RXFP1) und Effektormolekül (MMP-2) ist es wahrscheinlich, dass CMT über das Relaxin eine autonome Steuerung ihrer Invasivität vornehmen können. RXFP1 scheint dabei eine Regulationsfunktion in der Relaxinsensibilität der CMT-Zelle zuzukommen, die in Zukunft durch die Messung der RXFP1-Expressionsintensität prognostisch nutzbar sein könnte. Zudem ist RXFP1 im CMT damit auch ein möglicher Ansatzpunkt für eine neue, auf Relaxinanaloga basierende, antimetastatische Therapie, die bereits an humanen Tumorzellen und in Mausmodellen erprobt wird (FENG et al. 2007; HOSSAIN et al. 2010). Durch den Nachweis von Relaxin und RXFP1 im CMT und ihre wahrscheinliche Relevanz für die Metastasierung ergeben sich somit neue Möglichkeiten für eine exaktere Prognose und verbesserte antimetastatische Therapie von CMT sowie die Chance, den Hund als Modell für die Erforschung Relaxin basierter Therapien des humanen Brustkrebses zu nutzen. / Canine mammary tumours (CMT) are one of the main reasons of death in female dogs (SIMON et al. 2001). Due to its peripheral location, it is normally not the primary tumour, but its metastases, which are life-threatening as they often impair the function of vital organs, such as lung, heart or brain (CLEMENTE et al. 2010). Currently used techniques for the detection of metastasis can either barely describe invasive growth patterns or detect already existing metastases. Molecular markers to determine the metastatic potential early and reliably, before metastatic spreading has occurred, are still lacking. The peptide hormone relaxin is well known as an inductor of matrix metalloproteinases (MMP) in numerous physiological as well as pathological situations in humans (TOO et al. 1984; UNEMORI und AMENTO 1990; PALEJWALA et al. 2001; BINDER et al. 2002; KLONISCH et al. 2007; HENNEMAN et al. 2008). MMP are key-enzymes of connective tissue remodelling which is a prerequisite for metastasis (WOODHOUSE et al. 1997). In women, the plasma relaxin concentration is a marker for metastatic breast cancer (BINDER et al. 2004). However, in dogs, the concentration of circulating relaxin seems to have no diagnostic value (SCHWEIZER 2010). But, possibly relaxin is expressed locally in the canine mammary tissue as postulated by GOLDSMITH et al. (1994) and SILVERTOWN et al. (2003) and it could therefore act as a pro-invasive and pro-metastatic factor in an auto- or paracrine manner as it does in various human tumours (KLONISCH et al. 2007). Thus, the present study should examine the intratumoural expression of relaxin and its receptors as well as factors of connective tissue remodelling and evaluate their prognostic abilities. In two sections of the study (LAMP et al. 2009; LAMP et al. 2011), CMT-tissue samples from n=31 bitches (LAMP et al. 2009) and n=59 bitches (LAMP et al. 2011), respectively, were analysed for their expression of relaxin, its receptors RXFP1 and RXFP2, MMP-1, -2, -3, -9 and MMP-13 as well as the oestradiol receptors ERα and ERβ and the progesterone receptor (PR) using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). Additionally, the plasma concentrations of the hormones relaxin, oestradiol and progesterone were tested for possible connections with the local gene expression. In the second section of the study, the expression of relaxin and RXFP1 was also examined immunohistologically in n=9 CMT tissue samples (LAMP et al. 2011) and the prognostic value of all parameters examined was assessed by a multivariate analysis. The expression analysis showed that CMTs are a novel site of expression of relaxin and its main receptor RXFP1 in the dog. These results were confirmed by the immunohistological examination. Moreover, the immunohistological analysis demonstrated that relaxin seems to be expressed mainly in myoepithelial cells. However, the strongest signals for RXFP1 were located in epithelial cells of the CMT, thus RXFP1 is colocalised with the expression of MMP-2 and MMP-9 reported in epithelial CMT-cells (PAPPARELLA et al. 1997; HIRAYAMA et al. 2002; PAPPARELLA et al. 2002). The quantitative expression analysis revealed correlations of expression intensities for relaxin, RXFP1 and MMP-2. The expression of RXFP1 presented as an independent marker for metastasis with a 15-fold risk increase for patients with an expression intensity above the study-specific cut-off. All local gene expressions examined where independent from systemic plasma concentrations of relaxin, oestradiol and progesterone. The results propose an important role for intratumourally expressed relaxin as an auto- or paracrine modulator of MMP expression, which is important for invasiveness and metastasis. Due to the mRNA detection, the protein colocalisation of RXFP1 with MMP-2 and MMP-9 as well as the correlation of gene expressions of the ligand (relaxin), the receptor (RXFP1) and the effector (MMP-2) it is highly probable that CMT can autonomously regulate their invasiveness via locally expressed relaxin. RXFP1 seems to have a regulatory function in the relaxin responsiveness of CMT cells, which may be of prognostical use in the future. In addition, RXFP1 is also a possible target for a novel antimetastatic therapy based on relaxin analoga which has been tested in human tumour cells and mice (FENG et al. 2007; HOSSAIN et al. 2010). The detection of relaxin and RXFP1 in the CMT and their probable relevance for metastasis could be a basis for a more precise prognosis of CMT, improved anti-metastatic therapies in the dog and the use of the dog as a model for relaxin-based therapies of human breast cancer.

Le cancer épithélial de l'ovaire (EOC) a la particularité d'être diagnostiqué à un stade avancé chez 75% des patientes et de récidiver dans un grand nombre de cas malgré une bonne réponse initiale à la chimiothérapie, expliquant ainsi son pronostic sombre. Le rôle du microenvironnement tumoral semble être de premier plan dans le développement et la survie des cellules cancéreuses mais il existe encore peu de données concernant les cellules mésenchymateuses souches (MSC). Dans ce travail nous avons donc cherché à déterminer les mécanismes moléculaires entre les MSC et les cellules tumorales ovariennes. Dans la première partie de ce travail, nous avons mis en évidence l'émergence d'un profile pro-métastatique des cellules tumorales ovariennes après contact avec les MSC. Nous avons ensuite développé un modèle d'infiltration tumorale 3D révélant que les MSC augmentaient la dissémination tumorale ovarienne par la sécrétion d'IL6. Enfin nous avons démontré que les MSC étaient capables d'induire chez les cellules tumorales ovariennes un phénotype thermotolérant lié à la sécrétion CXCL12. Ces données vont donc toutes dans le même sens en démontrant les propriétés pro-tumorales des MSC et ouvrent de nouvelles perspectives de thérapies ciblant les interactions entre le stroma et la tumeur.

Background: Pancreatic métastases from renal cell carcinoma (RCC) are clinically rare but highly resectable. The aim of this article is to identify patients who profit from pancreatic resection of RCC despite the invasiveness of the surgery. Methods: Between January 1996 and December 2007, data from 744 patients were collected in a prospective pancreatic surgery database, and patients with metastasis into the pancreas from RCC were identified. Results: Resective surgery was performed in 14 patients with metastasis to the pancreas from RCC. Most patients were clinically asymptomatic. The median interval between primary treatment of RCC and occurrence of pancreatic metastasis was 94 months (range 32–158). The morbidity rate was 42.8%. Patients with a metastasis size <2.5 cm had a much better survival after resection (100 months) than those with a metastasis size >2.5 cm (44 months). Moreover, the number of métastases predicts the survival after resection. Conclusions: In patients with pancreatic métastases from RCC who have only limited disease, complete resection of all lesions can be successfully performed with a low rate of complications. Thus, patients with a history of RCC should be monitored for more than 10 years after nephrectomy to detect recurrence. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.