No description available.

No description available.

Les cellules de Sertoli (CS) jouent des rôles essentiels pour la régulation de la spermatogenèse, via la signalisation modulée par le récepteur aux androgènes (RA). Les objectifs de ce travail ont été d’identifier les mécanismes moléculaires de la régulation androgénique et des rôles des partenaires moléculaires dans la régulation androgénique des cellules de Sertoli pendant le développement testiculaire, du fœtus à l’âge adulte, pendant différentes stades du développement. Nous avons caractérisé et étudié une nouvelle lignée immortalisée, mature, de CS, ST38c, présentant une expression substantielle de RA endogène, une activation transcriptionnelle du RA induite par les androgènes, ainsi qu’une régulation et une stabilisation de la protéine RA par des mécanismes post traductionnels. Ce modèle a été utilisé afin de tester l’hypothèse que la suppression de l’hormone antimüllérienne (AMH) est modulée directement par les androgènes, via la RA, dans les cellules matures de Sertoli.En parallèle, nous avons testé l’hypothèse que la résistance physiologique aux androgènes du nouveau-né est liée à l’expression différentielle de quelques corégulateurs du RA. Ainsi, nous avons analysé l’expression et la contribution de deux corégulateurs du RA (SRC-2 et HBO1) pendant l’ontogenèse testiculaire humaine et pour des pathologies liées au disfonctionnement de l’action des androgènes ou du RA (syndromes d’insensibilité aux androgènes, hypogonadisme hypogonadotrophique congénital).Nous avons démontré, après des essais de transfection in vitro, que SRC-2 est un coactivateur, alors que HBO1 est in corepresseur du RA dans un modèle de cellules de Sertoli. Nous avons cartographié l’expression testiculaire humaine de SRC-2 et HBO1, pendant différentes stades du développement pré et postnatal et nous avons démontré que SRC-2 présente une expression stable, contrastant avec le profil d’expression différentielle, progressive avec l’âge de RA dans la cellule de Sertoli, suggérant que l’expression du SRC-2 est indépendante de la signalisation androgènique. Par contre, HBO1 et le RA présentent un profil de maturation et d’expression temporelle corrélé, suggérant que l’expression du HBO1 serait liée à une signalisation du RA fonctionnelle dans les cellules de Sertoli. Nous avons aussi démontré que l’expression du HBO1 est induite par les androgènes en présence du RA. Contrairement au SRC-2, HBO1 est non seulement exprimé dans les cellules de Sertoli, mais aussi dans les spermatogonies, pouvant représenter un marqueur potentiellement intéressant des cellules germinales.Enfin, nous avons aussi étudié l’immuno-expression testiculaire de RA et AMH chez des patients post pubères avec un syndrome de déficit en 5α-réductase type 2, et insensibilité minime aux androgènes (MAIS), afin d’étudier la contribution des androgènes (testostérone versus dihydrotéstosterone) pour la spérmatogenèse, la répression de l’AMH ainsi que pour mieux comprendre les perspectives de fertilité chez ces patients. / Sertoli cells (SC) have essential roles in the androgen regulation of spermatogenesis, via the androgen receptor (AR)-mediated signaling. This work aimed at identifying the molecular mechanisms related to the androgenic regulation of the AR and its molecular partners in Sertoli cells during different testicular developmental stages. We first characterized and studied a novel murine mature immortalized Sertoli cell line, called ST38c, which harbors substantial expression of endogenous AR, conserves its androgen-dependent transcriptional activation and exhibits agonist-dependent transcriptional and posttranslational regulation, as well as posttranslational stability.We used this cellular model in order to test the hypothesis that anti Müllerian Hormone (AMH) suppression in mature Sertoli cells would be directly androgen and AR mediated.Next, we hypothesized that the physiological androgen resistance in the neonate would also be related to the differential expression of several AR coregulators. Therefore, we analysed the differential expression and contribution of two AR co-regulators (SRC-2 and HBO1) during human testicular ontogeny, as well as in pathologies associated with androgen action or AR impairment (such as androgen insensitivity syndromes, congenital hypogonadotropic hypogonadism).Using in vitro transfection assays, we showed that SRC-2 is an AR coactivator while HBO1 is an AR corepressor in Sertoli cell models. We provided the cartography of SRC-2 and HBO1 expression during human testicular postnatal different stages and showed that SRC-2 presented a stable expression contrasting with the progressive evolution profile of the AR signaling, suggesting that Sertoli SRC-2 expression was independent of the androgen signaling. Interestingly, HBO1 and AR presented a temporal and positively correlated maturation profile, suggesting that HBO1 expression would be related to a functional AR signaling in the Sertoli cell. Moreover HBO1 expression is induced by androgens in the presence of the AR. Unlike SRC-2, HBO1 is not only expressed in Sertoli cells, but also in spermatogonia, being an interesting germ cell marker.Finally, we also studied AR and AMH immunoexpression in posptubertal cases of 5-α reductase type 2 deficiency and minimal androgen receptor resistance, in respect with the spermatogenesis status of seminiferous tubules, and androgen induced AMH suppression in order to assess the differential contribution of testosterone versus dihydrotestosterone and gather more information about the fertility perspectives in this particular pathologies.

This PhD aimed to elucidate the mechanisms by which polymorphisms may alter androgen-induced transactivation of androgen receptor (AR) target genes which may be important in prostate cancer aetiology. The second aspect of this PhD focused on identifying and characterising functional polymorphisms that may have utility as predictive risk indicators for prostate cancer and which may aid in earlier therapeutic intervention and better disease management. Analyses were carried out on the kallikrein-related peptidase 3 (KLK3), also known as the prostate specific antigen (PSA), gene and the kallikrein-related peptidase 4 (KLK4) gene. The PSA and KLK4 genes are part of the serine protease family that have trypsin or chymotrypsin like activity and are thought to play a role in the development of hormone-dependent cancers in tissues such as those in the prostate, breast, endometrium and ovaries. In the prostate, PSA is regulated by androgens and three androgen response elements (AREs) have been described in the promoter and upstream enhancer region. The PSA ARE I harbours a polymorphism at -158 bp from the transcription initiation site (TIS) that results in a G to A transition (G-158A). This PhD investigated the functional significance of the PSA G-158A polymorphism which has been reported to be associated with prostate cancer risk. Electromobility shift assays (EMSAs) investigating the interaction of ARE I variants with the AR DNA binding domain (AR-DBD) demonstrated that the A allele had a two-fold increased binding affinity for the AR-DBD when compared with the G allele. This was confirmed with endogenous AR in limited proteolysis-EMSA experiments. The limited proteolysis-EMSA experiments also demonstrated differential sensitivities of PSA ARE I alleles to trypsin digestion, which suggests that the G-158A polymorphism has an allosteric effect on the AR that alters AR/ARE I complex stability. Furthermore, Chromatin Immunoprecipitation (ChIP) assays suggest that the A allele more readily recruited the AR in vivo when compared with the G allele and is consistent with the in vitro binding data. Luciferase reporter assays carried out in both LNCaP and 22Rv1 prostate cancer cells, and using the natural (dihydrotestosterone; DHT) ligand demonstrated that the A allele was more responsive to androgens in LNCaP cells. Hence, this study has elucidated the potential mechanisms by which the G-158A polymorphism may differentially regulate PSA expression (of which up-regulation of PSA is thought to be important in prostate cancer development and progression). KLK4 has similar tissue-restricted expression as PSA and is up-regulated by steroid hormones in many endocrine cells including those in the prostate. A putative ARE (KLK4-pARE) located at -1,005 to -1019 relative to the more predominantly used transcription initiation site, TIS3, was initially found in supershift assays using AR antibodies to interact with endogenous AR. However, subsequent EMSA analysis using purified AR-DBD suggest that KLK4-pARE may be interacting with the AR indirectly. To investigate this hypothesis, a tandem construct of KLK4-pARE was cloned into the pGL3-Promoter vector for hormone-induced reporter assays. However, reporter assays did not demonstrate any responsiveness of KLK4-pARE to androgens, estradiol or progestins. Consequently, Real-Time PCR was carried out to reassess the hormonal regulation of KLK4 at the mRNA level. Consistent with the literature, data from this study suggests that KLK4 may be up-regulated by androgens, progestins and estradiol in a cyclical manner. Hormone-induced luciferase reporter assays were then carried out on seven promoter constructs that span 2.8 kb of the KLK4 promoter from TIS3. However, none of the seven promoter constructs demonstrated any significant responsiveness to androgens, estradiol or progestins. This study suggests that hormone response elements (HREs) that may drive the hormonal regulation of KLK4 in prostate cancer may be located further upstream from the promoter region investigated in this PhD, or alternatively, may lie 3' of TIS3. The characterisation of KLK4 promoter polymorphisms and their flanking sequences were also carried out in parallel to the functional work with the intent to assess the functional significance of any polymorphisms that may be located within HREs. In total 19 polymorphisms were identified from the public databases and from direct sequencing within 2.8 kb of the KLK4 promoter from TIS3. However, the functional and clinical significance of these 19 polymorphisms were not further pursued given the negative findings from the functional work. The PSA AR enhancer region was also assessed for potential polymorphisms that may be associated with prostate cancer risk. A total of 12 polymorphisms were identified in the PSA enhancer of which two (A-4643G and T-5412C) have been reported to alter functionality of the enhancer region and thus, prioritised for further analysis. Association analysis for prostate cancer risk was then carried out on these PSA enhancer polymorphisms as none of the KLK4 promoter polymorphisms were found in functional HREs. No significant association for either the A-4643G or T-5412C polymorphism with prostate cancer risk was found at the P = 0.05 level. However, under an age-adjusted dominant model a 1.22- (95% CI = 1.16-1.26) and 1.23-fold (95% CI = 1.17-1.29) increased risk for prostate cancer was found for the A-4643G or T-5412C polymorphisms, respectively. Both polymorphisms were also assessed for association with tumour grade and stage and PSA levels. Genotypes were significantly different for the A-4643G and T-5412C polymorphisms with tumour stage and PSA levels, respectively. However, these results are likely to be biased by the case population which consist primarily of men who presented with incidental (pT1) and organ-confined (pT2) tumours. To summarise, the A-4643G and T-5412C polymorphisms are unlikely to be associated with prostate cancer risk, PSA levels or stage/grade of disease. However, further analyses in a larger cohort is warranted given that these polymorphisms alter androgen responsiveness of the PSA enhancer and that elevated PSA levels are indicative of men with prostate cancer. To summarise, this PhD has elucidated the functional significance of the PSA G-158A polymorphism in prostate cancer and which may be important in prostate cancer patho-physiology. This PhD has also furthered the understanding of the hormonal regulation of KLK4 in prostate cancer cells. Finally, this PhD has carried out a pilot study on two functional PSA enhancer polymorphisms (A-4643G and T-5412C) with prostate cancer risk.

Dietary supplements sold for anabolic benefits or performance enhancement often contain substances, which are nonapproved and might lack quality controls. With regard to athletes, the inclusion of substances or methods in the prohibited list of the World Anti-Doping Agency is based on medical or scientific evidence. 5α-hydroxy-laxogenin is a synthetic spirostane-type steroid, which is contained in dietary supplements and advertised as anabolic agent. To date, evidence is missing on anabolic or androgenic activity of 5α-hydroxy-laxogenin. We investigated its androgenic potential in two in vitro bioassays. While no activity was observed in the yeast androgen screen, 5α-hydroxy-laxogenin was able to trans-activate the androgen receptor in human prostate cells in a dose-dependent manner. Interestingly, a biphasic response was observed with antagonistic properties at lower concentrations and agonistic effects at higher concentrations tested. The demonstrated androgenic properties of the higher concentrations demonstrate that further investigations should focus on the safety as well as on potential anabolic effects of 5α-hydroxy-laxogenin. This is of interest with regard to abuse for doping purposes.

Les cancers moléculaires apocrines sont un sous-groupe de cancer du sein caractérisé par l'expression du récepteur aux androgènes (RA), l'absence du récepteur aux oestrogènes (RE) et l'expression paradoxale de nombreux gènes typiquement exprimés dans les tumeurs RE positives. Une proportion significative de ces patientes va récidiver sous forme de métastases dont la prise en charge repose sur des traitements non spécifiques (chimiothérapies). En préclinique, la lignée cellulaire MDA-MB-453 a été identifiée comme ayant un profil transcriptomique similaire à ce sous-groupe tumoral. En clinique, les essais réalisés dans ce sousgroupe tumoral avec différents anti-androgènes, dont l’abiratérone (inhibiteur de la synthèse des androgènes), retrouvent un bénéfice clinique chez environ 25% des patientes. L’objectif de cette thèse est d’améliorer les connaissances et les prises en charge thérapeutiques spécifiques de ces tumeurs. Nos données précliniques comparatives montrent que l'ODM-201, nouvel antiandrogène, ne présente pas une efficacité supérieure par rapport aux antiandrogènes déjà étudiés. Afin de contourner les limites des lignées cellulaires identifiées dans ce premier projet, nous avons démontré la nécessité de développer de nouveaux modèles : les Patient-Derived-Xenograft orthotopiques. Notre deuxième projet est en faveur d’une meilleure sélection des patientes à traiter par abiratérone notamment basé sur des caractéristiques immunohistochimiques apocrines. Chez les patientes ne présentant pas ces caractéristiques, nous avons isolé CHEK1 comme une cible d’intérêt en combinaison thérapeutique pour majorer les taux de réponse de l’abiratérone en monothérapie. / Molecular apocrine cancers are a subgroup of breast cancer characterized by the expression of the androgen receptor (AR), the absence of the estrogen receptor (ER) and the paradoxical expression of many genes typically expressed in ER positive tumors. A significant proportion of these patients will recur in the form of metastases whose management is based on non-specific treatments (chemotherapy). In preclinical study, the MDA-MB-453 cell line was identified as having a transcriptomic profile similar to this tumor subgroup. Clinical trials in this tumor subgroup testing different antiandrogens, including abiraterone (inhibitor of androgen synthesis), found a clinical benefit in about 25% of patients. The aim of this thesis is to improve the knowledge and the specific therapeutic management of these tumors. Our comparative preclinical data show that ODM-201, a new anti-androgen, does not show superior efficacy compared to previously studied anti-androgens. In order to circumvent the limits of cell lines provided by this first project, we have shown the need to develop new models: orthotopic Patient-Derived-Xenograft. Our second project favors a better selection of patients to be treated with abiraterone, especially based on apocrine immunohistochemical characteristics. In patients without these characteristics, we isolated CHEK1 as a target of interest in combination therapy to increase response rates of abiraterone monotherapy.

Introduction: We investigated the health-related quality of life (HRQoL) of long-term prostate cancer patients who received leuprorelin acetate in microcapsules (LAM) for androgen-deprivation therapy (ADT). Methods: The observational study was carried out by 30 office-based German urologists in 536 prostate cancer (PCa) patients treated for ≥5 years with LAM and in 116 patients of an age-matched control group (CG). Data on HRQoL and health status was collected prospectively using validated questionnaires QLQ-C30, QLQ-PR25 and Karnofsky Index. Data on effectiveness (clinical response, prostate specific antigen [PSA], testosterone) and safety was collected retrospectively from patients’ health records. We used descriptive statistics to analyze the data. Results: The mean treatment duration was 8.6 years (range 4.5–19.8 years). General health status (QLQ-C30) was comparable for both groups. Differences were observed regarding physical – and role functioning. ADT patients rated single items slightly worse than CG. Karnofsky-Index showed comparable high values (median of 90%). QLQ-PR25 revealed more PCa-related symptoms for ADT patients. Within 6 months, median PSA level declined >90% and median testosterone levels declined below castration level from 4.0 to 0.2 ng/mL. Clinical response (European Organisation for Research and Treatment of Cancer criteria) was observed in at least 90% of ADT patients. Conclusions: Long-term ADT with LAM is a well-accepted, tolerated, effective, and low-burden treatment option for patients with advanced, hormone-sensitive PCa.

No description available.

No description available.

The locomotor activity of the reindeer is separated into a diurnal and a nocturnal phase which, in turn, consists of a  number of short-term activity bursts. The onset and termination  of diurnal and nocturnal activity are largely in synchrony  with sunrise and sunset. Since the diurnal phase is  longer, total activity is dependent on the photoperiod. Total  activity as well as the number of activity bursts is greater  in Jùne with continuous daylight than in December with 6  hours daylight. In winter, reindeer spent less time feeding  but more time ruminating and resting *than in summer. Synchronization  between individuals was also greatest in winter. It  is concluded that the described changes in the activity  pattern are due to an endogenous component and that this component  further controls food consumption and energy balance  of the reindeer.  Seasonal variations in behavioural patterns are described.  Several specific rutting behavioürs, sexual and agonostic,  emerge in the male during the prerut and persist until after  the rut. The preorbital gland is enlarged and secretory activity  is evident. The rut is cha'tabterized by the sudden appearance  of an odour in the breath of mature males followed a few  later by a strong odour in the urine. These odours persist for  a short time during the most intense period of rut. The amount  of androgen rizes sharply and reaches peak concentrations in  late September - early October, decreasing thereafter. The  described behavioural cues occur with a certain constancy in  time in concecutive seasons.  A quantification of the amounts of secretory epithelia in preorbital,  interdigital, caudal and tarsal glands reveals that  only the preorbital gland is subjected to any seasonal and  sexual variation. The apocrine epithelium in this gland is  most developed in mature males during the rutting season and  seems to be dependent upon the presence of an elevated androgen  concentration. The tarsal gland is the least developed  gland while the interdigital and caudal glands are more welldeveloped  and structurally complex. Scattered apocrine glands  are only developed on the legs, the ventral body, oral angle  and in the rump patch. / digitalisering@umu