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MEN1 mutation predisposes patients to multiple endocrine neoplasia type 1 (MEN1), a genetic syndrome associated with the predominant co-occurrence of endocrine tumors. Intriguingly, recent evidence has suggested that MEN1 could also be involved in the development of breast and prostate cancers, two major hormone-related cancers. The first clues as to its possible role arose from the identification of the physical and functional interactions between the menin protein, encoded by MEN1, and estrogen receptor α and androgen receptor. In parallel, our team observed that aged heterozygous Men1 mutant mice developed cancerous lesions in mammary glands of female and in the prostate of male mutant mice at low frequencies, in addition to endocrine tumors. Finally, observations made both in MEN1 patients and in sporadic breast and prostate cancers further confirmed the role played by menin in these two cancers. In this review, we present the currently available data concerning the complex and multifaceted involvement of MEN1 in these two types of hormone-dependent cancers.

IntroductionProstate multiparametric MRI (mpMRI) has shown good sensitivity in detecting cancers with an International Society of Urological Pathology (ISUP) grade of ≥2. However, it lacks specificity, and its inter-reader reproducibility remains moderate. Biomarkers, such as the Prostate Health Index (PHI), may help select patients for prostate biopsy. Computer-aided diagnosis/detection (CAD) systems may also improve mpMRI interpretation. Different prototypes of CAD systems are currently developed under the Recherche Hospitalo-Universitaire en Santé / Personalized Focused Ultrasound Surgery of Localized Prostate Cancer (RHU PERFUSE) research programme, tackling challenging issues such as robustness across imaging protocols and magnetic resonance (MR) vendors, and ability to characterise cancer aggressiveness. The study primary objective is to evaluate the non-inferiority of the area under the receiver operating characteristic curve of the final CAD system as compared with the Prostate Imaging-Reporting and Data System V.2.1 (PI-RADS V.2.1) in predicting the presence of ISUP ≥2 prostate cancer in patients undergoing prostate biopsy.MethodsThis prospective, multicentre, non-inferiority trial will include 420 men with suspected prostate cancer, a prostate-specific antigen level of ≤30 ng/mL and a clinical stage ≤T2 c. Included men will undergo prostate mpMRI that will be interpreted using the PI-RADS V.2.1 score. Then, they will undergo systematic and targeted biopsy. PHI will be assessed before biopsy. At the end of patient inclusion, MR images will be assessed by the final version of the CAD system developed under the RHU PERFUSE programme. Key secondary outcomes include the prediction of ISUP grade ≥2 prostate cancer during a 3-year follow-up, and the number of biopsy procedures saved and ISUP grade ≥2 cancers missed by several diagnostic pathways combining PHI and MRI findings.Ethics and disseminationEthical approval was obtained from the Comité de Protection des Personnes Nord Ouest III (ID-RCB: 2020-A02785-34). After publication of the results, access to MR images will be possible for testing other CAD systems.Trial registration numberNCT04732156.

Background Recent studies highlighted the increased frequency of AR-low or -negative prostate cancers (PCas) and the importance of AR-independent mechanisms in driving metastatic castration-resistant PCa (mCRPC) development and progression. Several previous studies have highlighted the involvement of the MEN1 gene in PCa. In the current study, we focused on its role specifically in AR-independent PCa cells. Methods Cell tumorigenic features were evaluated by proliferation assay, foci formation, colony formation in soft agar, wound healing assay and xenograft experiments in mice. Quantitative RT-PCR, Western blot and immunostaining were performed to determine the expression of different factors in human PCa lines. Different ChIP-qPCR-based assays were carried out to dissect the action of JunD and β-catenin. Results We found that MEN1 silencing in AR-independent cell lines, DU145 and PC3, resulted in an increase in anchorage independence and cell migration, accompanied by sustained MYC expression. By searching for factors known to positively regulate MYC expression and play a relevant role in PCa development and progression, we uncovered that MEN1-KD triggered the nuclear translocation of JunD and β-catenin. ChIP and 3C analyses further demonstrated that MEN1-KD led to, on the one hand, augmented binding of JunD to the MYC 5′ enhancer and increased formation of loop structure, and on the other hand, increased binding of β-catenin to the MYC promoter. Moreover, the expression of several molecular markers of EMT, including E-cadherin, BMI1, Twist1 and HIF-1α, was altered in MEN1-KD DU145 and PC3 cells. In addition, analyses using cultured cells and PC3-GFP xenografts in mice demonstrated that JunD and β-catenin are necessary for the altered tumorigenic potential triggered by MEN1 inactivation in AR-independent PCa cells. Finally, we observed a significant negative clinical correlation between MEN1 and CTNNB1 mRNA expression in primary PCa and mCRPC datasets. Conclusions Our current work highlights an unrecognized oncosuppressive role for menin specifically in AR-independent PCa cells, through the activation of JunD and β-catenin pathways.

Dysregulated androgen receptor (AR) plays a crucial role in prostate cancer (PCa) development, though further factors involved in its regulation remain to be identified. Recently, paradoxical results were reported on the implication of the MEN1 gene in PCa. To dissect its role in prostate luminal cells, we generated a mouse model with inducible Men1 disruption in Nkx3.1-deficient mice in which mouse prostatic intraepithelial neoplasia (mPIN) occur. Prostate glands from mutant and control mice were analyzed pathologically and molecularly; cellular and molecular analyses were carried out in PCa cell lines after MEN1 knockdown (KD) by siRNA. Double-mutant mice developed accelerated mPIN and later displayed microinvasive adenocarcinoma. Markedly, early-stage lesions exhibited a decreased expression of AR and its target genes, accompanied by reduced CK18 and E-cadherin expression, suggesting a shift from a luminal to a dedifferentiated epithelial phenotype. Intriguingly, over 60% of menin-deficient cells expressed CD44 at a later stage. Furthermore, MEN1 KD led to the increase in CD44 expression in PC3 cells re-expressing AR. Menin bound to the proximal AR promoter and regulated AR transcription via the H3K4me3 histone mark. Interestingly, the cell proliferation of AR-dependent cells (LNCaP, 22Rv1, and VCaP), but not of AR-independent cells (DU145, PC3), responded strongly to MEN1 silencing. Finally, menin expression was found reduced in some human PCa. These findings highlight the regulation of the AR promoter by menin and the crosstalk between menin and the AR pathway. Our data could be useful for better understanding the increasingly reported AR-negative/NE-negative subtype of PCa and the mechanisms underlying its development.

Proof of the clinical utility of a biomarker is when its use informs a management decision and improves patient outcomes relative to when it is not used.To model the clinical benefit of the nuclear-localized androgen receptor splice variant 7 (AR-V7) test for men with progressing metastatic castration-resistant prostate cancer (mCRPC) at the second line of therapy or greater to inform the choice of an androgen receptor signaling inhibitor (ARSI) or a taxane.The study population was a cross-sectional cohort of 193 unique patients with progressing mCRPC from whom 255 samples were drawn at the time of the second line or later treatment decision who then received an ARSI or taxane, with up to 3 yr of additional follow-up Circulating tumor cells (CTCs) were identified from blood samples and tested for AR-V7. Physicians were blinded to AR-V7 status and the testing laboratory was blinded to outcomes.We measured physician propensity for choosing an ARSI or taxane based on patient prognosis. We also measured overall survival (OS) adjusted for physician propensity by drug class; OS data were analyzed both without and with knowledge of nuclear-localized AR-V7 status.Treating physicians had a propensity for choosing a taxane over an ARSI for patients with more advanced disease or who received an ARSI as the immediate prior therapy. After adjusting for physician propensity, discernible OS differences were not observed between taxane- and ARSI-treated patients (median 15.6 vs 14.4 mo; p =0.11). Patients with detectable nuclear-localized AR-V7 in CTCs had superior survival with taxanes over ARSIs (median 9.8 vs 5.7 mo; p = 0.041). AR-V7-negative patients had superior survival on ARSIs over taxanes (p = 0.033) but overlapping curves limit the interpretation. Mutivariable models showed a robust interaction between AR-V7 status and drug, and a lower risk of death on taxanes for AR-V7-positive men.Use of the nuclear-localized AR-V7 CTC test to inform treatment choice can improve patient outcomes relative to decisions based solely on standard-of-care measures.Men with metastatic prostate cancer who test positive for AR-V7 protein in circulating tumor cells are likely to live longer if taxane chemotherapy is used.

International audience; Steroid receptors (SRs) are members of the nuclear receptor family, which are ligand-activated transcription factors. SRs regulate many physiological functions including development and reproduction, though they can also be involved in several pathologies, especially cancer. Highly controlled cellular responses to steroids involve transcriptional regulation (genomic activity) combined with direct activation of signaling cascades (non-genomic activity). Non-genomic signaling has been extensively studied in cancer, mainly in breast cancer for ER and PR, and prostate cancer for AR. Even though most of the studies have been conducted in cells, some of them have been confirmed in vivo, highlighting the relevance of this pathway in cancer. This review provides an overview of the current and emerging knowledge on non-genomic signaling with a focus on breast and prostate cancers and its clinical relevance. A thorough understanding of ER, PR, AR and GR non-genomic pathways may open new perspectives for the development of therapeutic strategies.

Background In metastatic castration-resistant prostate cancer (mCRPC), androgen receptor splice variant 7 (AR-V7) expression is associated with a low response to androgen receptor signaling (ARS) inhibitors such as abiraterone or enzalutamide. Objective To perform a highly sensitive assay for detecting AR-V7 (hsAR-V7) in circulating tumor cells (CTCs) and evaluate its ability to predict response to ARS inhibitors. Design, setting, and participants From 41 mCRPC patients, CTCs were prospectively enriched using AdnaTest platform and analyzed for AR-V7 with and without the highly sensitive assay. Outcome measurements and statistical analysis The first objective of the study was to compare AR-V7 detection rates with and without the highly sensitive assay. Next, we investigated how AR-V7 (detected without the highly sensitive assay) and hsAR-V7 status influenced prostate-specific antigen (PSA) response and long-term clinical outcomes (PSA progression-free survival [PFS] and radiological PFS) after ARS-inhibitor treatment. Finally, discriminatory abilities of the assays were assessed by C-index to compare their impact on long-term clinical outcomes. Results and limitations AR-V7 detection rates increased from 22% to 56% when the highly sensitive assay was used. The discriminatory abilities of hsAR-V7 for PSA PFS (C-index, 0.74; 95% confidence interval [CI], 0.60–0.88) and radiological PFS (0.70; 95% CI, 0.55–0.85) were higher than those of AR-V7 detected without the highly sensitive assay (0.60, 0.51–0.72, and 0.56, 0.44–0.67, respectively). After ARS-inhibitor treatment, PSA response was lower in hsAR-V7+ (53%) than in hsAR-V7– (93%) patients (p = 0.016). AR-V7+ patients had shorter median PSA PFS (3.0 vs 10.6 mo, p = 0.032) and nonsignificantly shorter median radiological PFS (6.0 vs 14.8 mo, p = 0.24) compared with AR-V7– patients. The hsAR-V7+ status was associated with shorter median PSA PFS (3.0 mo vs not reached, p = 0.0001) and radiological PFS (median, 6.0 mo vs not reached, p = 0.0026). Conclusions The hsAR-V7 assay achieved the highest AR-V7 detection rates among those reported in mCRPC. Discriminatory abilities for long-term clinical outcomes were better with hsAR-V7 assay. Patient summary We prospectively analyzed circulating tumor cells from men with metastatic castration-resistant prostate cancer for androgen receptor splice variant 7 (AR-V7) status using a highly sensitive assay. It yielded higher AR-V7 detection rates and predicted resistance to androgen receptor signaling inhibitors with better discriminatory abilities for long-term clinical outcomes.

INTRODUCTION AND OBJECTIVES:A 2-gene, urine-based molecular test that combines mRNA biomarkers with clinical factors can risk stratify patients for high-grade prostate cancer (PCa). To ensure the g...

A 2-gene, urine based molecular test that combines mRNA biomarkers with clinical factors can risk stratify patients for clinically significant prostate cancer. To ensure the generalizability of assay results we optimized and validated the clinical model for men with serum prostate specific antigen less than 10 ng/ml who were undergoing initial prostate biopsy.Urine samples were collected from 1,955 men from The Netherlands, France and Germany prior to an initial prostate biopsy and study subjects were divided into training and validation cohorts. Urinary HOXC6 and DLX1 mRNA levels were quantified and RNA results were then combined with other risk factors in a clinical model optimized to detect ISUP (International Society of Urological Pathology) Grade Group 2 or greater prostate cancer in men with prostate specific antigen less than 10 ng/ml. Results in the validation cohort were compared with the PCPTRC (Prostate Cancer Prevention Trial Risk Calculator), version 2.0.The optimal clinical model included urinary HOXC6 and DLX1 mRNA levels, patient age, digital rectal examination and prostate specific antigen density (serum prostate specific antigen/prostate volume). In the 715 validation cohort subjects with prostate specific antigen less than 10 ng/ml the AUC was 0.82 with 89% sensitivity, 53% specificity and 95% negative predictive value. The PCPTRC AUC was 0.70. The full validation cohort of 916 men including all prostate specific antigen levels yielded an AUC of 0.85 with 93% sensitivity, 47% specificity and 95% negative predictive value. The PCPTRC AUC was 0.76.The 2-gene based urine assay, which is optimized for biopsy naïve patients with serum prostate specific antigen less than 10 ng/ml, demonstrated high sensitivity and negative predictive value to detect clinically significant prostate cancer. These data support using the test to help guide initial prostate biopsy decisions.

Low specificity of prostate-specific antigen results in a considerable number of unnecessary prostate biopsies in current practice. SelectMDx® predicts significant prostate cancer upon biopsy and is used to reduce the number of unnecessary initial prostate biopsies. Furthermore, potential overtreatment of insignificant prostate cancer can be reduced. Besides the diagnostic accuracy of the test, also the context in a specific country determines the potential health benefit and cost-effectiveness. Therefore, the health benefit and cost-effectiveness of SelectMDx were assessed in France, Germany, Italy, and Spain. A decision model was used to compare the current standard of care in which men undergo initial prostate biopsy in case of an elevated prostate-specific antigen, to a strategy in which SelectMDx was used to select men for biopsy. Model inputs most relevant to each of the four countries were obtained. With use of the model long-term quality-adjusted life years (QALYs) and healthcare costs were calculated for both strategies. In all four countries, the SelectMDx resulted in QALY gain and cost savings compared with the current standard of care. In France, SelectMDx resulted in 0.022 QALYs gained and cost savings of €1217 per patient. For Germany, the model showed a QALY gain of 0.016 and a cost saving of €442. In Italy, the QALY gain and cost savings were 0.031 and €762. In Spain 0.020 QALYs were gained and €250 costs were saved. The results of the model showed that with SelectMDx, QALYs could be gained while saving healthcare costs in the initial diagnosis of prostate cancer. The significant presence of overtreatment in the current standard of care in all four countries was the main factor that resulted in the beneficial outcomes with SelectMDx.

Resume But Evaluer la capacite du test urinaire PCA3 a predire le resultat des biopsies prostatiques dans une large cohorte issue du centre hospitalier Lyon-Sud. Patients et methodes Le score PCA3 a ete determine prospectivement chez 1015 patients adresses pour biopsies prostatiques. La capacite predictive du score PCA3 a ete evaluee par la comparaison des aires sous les courbes ROC, de modeles de regression logistique et une analyse par decision curve analysis (DCA). Resultats Le score PCA3 median etait significativement plus eleve chez les patients avec biopsies positives. L’AUC etait de 0,76, significativement plus elevee que celle du PSA a 0,55. Au seuil de 35, la sensibilite etait de 68 %, la specificite de 71 %, les valeurs predictives positive et negative de 67 % et 71 %, et l’efficience de 69 %. En analyse multivariee, le score PCA3 etait predicteur independant du resultat des biopsies et son addition a un modele de base comportant les donnees clinico-biologiques classiques apportait un gain diagnostique significatif. Au seuil de 20, pres de la moitie des biopsies a posteriori inutiles auraient ete evitees, tout en ayant ignore 7 % des cancers avec score de Gleason ≥ 7. Le score PCA3 n’apparaissait pas correle au score de Gleason, mais etait bien correle au volume tumoral (evalue par la proportion de carottes envahies). Conclusion Le test urinaire PCA3 presente des performances diagnostiques elevees pour le diagnostic precoce du CaP. Sa correlation avec l’agressivite du cancer s’exprime a travers le volume tumoral plus que par le score de Gleason. Niveau de preuve 5.

Summary Aim To evaluate the performance of urinary PCA3 test to predict prostate biopsy outcome in a large French cohort. Patients and methods A urine sample was prospectively obtained from 1015 patients undergoing prostate biopsies to determine the PCA3 score. The predictive value of PCA3 was explored using receiver operating characteristic curve analysis (ROC), multivariable logistic regression analysis and decision curve analysis. Results The median PCA3 score was significantly higher in patients with positive biopsies. The PCA3 score AUC was 0.76 (0.73–0.79), significantly higher than that of PSA (0.55; 0.51–0.58). At the cutoff of 35, sensitivity was 68%, specificity 71%, positive and negative predictive values 67% and 71%, and accuracy 69%. Using multivariate analysis, PCA3 score appeared as an independent predictor of biopsy outcome and its addition to a base model including usual clinicobiological parameters resulted in a significant increase in predictive accuracy. At the cutoff of 20, about 1/2 of the ultimately unnecessary biopsies would have been avoided while ignoring 7% of cancers with Gleason score ≥ 7. PCA3 score did not correlate with Gleason score but did correlate with tumor volume (proportion of positive cores). Conclusion Urinary PCA3 is a useful test with high diagnostic performances for early prostate cancer diagnosis. Its correlation with cancer aggressiveness seems rather represented by a link to prostate volume than Gleason score. Level of evidence 5.

Extracellular vesicles (EVs), especially exosomes, are now well recognized as major ways by which cancer cells interact with each other and stromal cells. The meaningful messages transmitted by the EVs are carried by all components of the EVs, i.e., the membrane lipids and the cargo (DNAs, RNAs, microRNAs, long non-coding RNAs, proteins). They are clearly part of the armed arsenal by which cancer cells obtain and share more and more advantages to grow and conquer new spaces. Identification of these messages offers a significant opportunity to better understand how a cancer occurs and then develops both locally and distantly. But it also provides a powerful means by which cancer progression can be detected and monitored. In the last few years, significant research efforts have been made to precisely identify how the EV trafficking is modified in cancer cells as compared to normal cells and how this trafficking is altered during cancer progression. Prostate cancer has not escaped this trend. The aim of this review is to describe the results obtained when assessing the meaningful content of prostate cancer- and stromal-derived EVs in terms of a better comprehension of the cellular and molecular mechanisms underlying prostate cancer occurrence and development. This review also deals with the use of EVs as powerful tools to diagnose non-indolent prostate cancer as early as possible and to accurately define, in a personalized approach, its present and potential aggressiveness, its response to treatment (androgen deprivation, chemotherapy, radiation, surgery), and the overall patients' prognosis.

Bexarotene (Targretin), approved since 1999 as a second-line treatment for late stage cutaneous T-cell lymphomas, has been shown to induce significant hypothyroidism through TSH suppression. This review revisits, through a case report, mechanisms by which rexinoids repress the expression ofTSHBgene as well asαTSHandTRHgenes. It appears that rexinoids suppress TSH independently from tri-iodothyronine. Bexarotene also differently affects the gene expression of deiodinases 1 and 2 as well as the peripheral clearance of thyroxine. These data might open new ways of research on the potential interaction between thyroid axis and endogenous rexinoids.

Resume Objectifs Determiner si l’ajout du score PCA3, et/ou de l’IRM prostatique permet d’ameliorer la selection des patients candidats a une premiere serie de biopsies de prostate. Methodes Une IRM prostatique multiparametrique (IRM MP) et un dosage de PCA3 ont ete realises avant biopsie chez les patients ayant une indication de premiere biopsie pour un toucher rectal anormal et/ou un PSA superieur a 4 ng/mL. Les sequences T2, dynamiques et de diffusion recherchaient une cible suspecte cotee sur un score de 4. Les patients ont ete inclus de facon prospective dans un seul centre. Les performances du PCA3 urinaire et de l’IRM ont ete evaluees par comparaison aux resultats des biopsies. Resultats Soixante-huit patients ont ete inclus, le PSA median etait de 5,2 ng/mL (3,2–28). La valeur predictive negative (VPN) des sores IRM 0, 1 et 2 etait respectivement de 80 %, 43 % et 69 %. La valeur predictive positive (VPP) des scores IRM de 3 et 4 etait de 50 % et 81 %. La valeur seuil discriminante pour le PCA3 etait de 21 (Se : 0,91 ; Sp : 0,50). Un seul patient avec des biopsies positives (0,5 mm de score de Gleason 3 + 3) avait a la fois une IRM negative et un PCA3 inferieur a 21. Conclusion L’association de l’IRM MP et du PCA3 permettait, dans cette etude, d’envisager une reduction du nombre de primo-biopsies inutiles sans ignorer la presence d’une eventuelle tumeur agressive.

To systematically review the evidence for the use of PSA and other biomarkers in the early detection of prostate cancer, we searched PubMed for clinical trials and studies assessing PSA and other biomarkers in the early detection of prostate cancer, published between 2000 and May 2013 that included >200 subjects. The level of evidence (LOE) for clinical utility was evaluated using the tumor marker utility grading system. A total of 84 publications, corresponding to 70 trials and studies were selected for inclusion in this review. We attributed a level of evidence (LoE) of IA to PSA for early PCa detection, but we do not recommend its use in mass screening. Emerging biomarkers were assessed in prospective case-control and cohort studies: PCA3 (n=3); kallikreins (n=3); [-2]proPSA (n=5); fusion oncogenes (n=2). These studies used biopsy results for prostate cancer to determine specificity and sensitivity, but they did not assess the effect on PCa mortality. The LoE attributed was III-C. PSA can be used for early prostate cancer detection but mass screening is not recommended. Studies on other biomarkers suggest that they could be used, individually or in combination, to improve the selection of patients with elevated PSA levels for biopsy, but RCTs assessing their impact on prostate cancer management and mortality are needed. A better use of available tests is possible for men at risk in order to maximize the risk-benefit ratio.

We describe a technique of S isotope analysis in sulfate form with the first separation stage involving anion-exchange and the second stage of mass-spectrometric analysis by MC-ICP-MS using standard-sample-standard bracketing. Ammonium in 1 : 1 stoichiometric proportion with sulfate was used to improve transmission and stability and to avoid cone and membrane clogging by condensable species. The working resolution of similar to 9000 allowed the main interferences, notably (SH)-S-32 on S-33, to be resolved. The matrix effect caused by phosphorus present in biological samples is negligible for S/P ratios \textgreater= 10: our chemical protocol allows S/P \textgreater= 150 to be routinely achieved. Replicate measurements of S standard solutions give values of isotopic abundances within errors of accepted values and demonstrate a reproducibility of +/- 0.10 parts per thousand for delta S-34 and +/- 0.15 parts per thousand for delta S-33 (2s). The technique is adequate for quantities as small as 10 nanomoles. We investigated the delta S-34 of 110 samples of cancer patients and 10 samples of rheumatoid arthritis patients. We avoided the use of blood collection tubes with sulfate-containing heparin. Sulfur in serum is transported by albumin and fibrinogen. Most serum and plasma delta S-34 values fall within a narrow interval of similar to 1 parts per thousand around a mean delta S-34(VCDT) of similar to 6.0 parts per thousand. The delta S-34 values of total blood, serum, and plasma are very similar. Despite the short turnover time of albumin and fibrinogen, S is surprisingly well regulated. Subtle variations of 0.2-0.3 parts per thousand around the mean value can be assigned to sex and age, with sulfur in male and adult samples tending to be heavier than in their female and juvenile counterparts. This narrow range of variations across the spectrum of a large number of individuals not selected for controlled dietary habits seems paradoxical. In general, breast and prostate cancer and rheumatoid arthritis have very little effect on the average serum delta S-34, but increase the scatter of values. We confirm that the serum of patients affected by liver cancer and other pathologies is depleted of albumin-born sulfur. While sulfur in the serum of patients with non-malignant liver pathologies tends to be isotopically light, the serum delta S-34 of medicated hepatocellular carcinoma patients tends to be at the high end of control values.

BACKGROUND The influence of chronic prostatitis on serum PSA level is well known. Whether it also influences potential new biomarkers of prostate cancer (PCa) has to be determined. We conducted a prospective study to evaluate the effect of chronic prostatitis on the PCa urinary marker PCA3. METHODS Included were 38 patients, mean-aged of 37.5 years, with clinical suspicion of chronic prostatitis. A simplified version of the Meares–Stamey four-glass localization test was performed and urine specimens were collected for cytological analysis and culture. A postprostatic massage urine sample was used for the urinary PCA3 test. RESULTS Four patients had an eventual diagnosis of urethritis and all had a PCA3 score less than 5. Among the remaining 34 patients, 7 had bacterial chronic prostatitis (NIH II prostatitis), 11 had abacterial chronic prostatitis (NIH IIIa), and 16 had non inflammatory prostatodynia (NIH IIIb). All these patients had a PCA3 score less than 28, that is, under the cutoff of 35, which is commonly used for prostate cancer diagnosis. Patients with NIH category IIIa prostatitis had significantly higher number of leukocytes and red cells as well as prostate cells in urine samples but their PCA3 scores did not differ from those of other prostatitis patients. CONCLUSION In this study, NIH II and III chronic prostatitis did not influence the PCA3 score. Our results suggest that increased PCA3 score is unlikely to be explained by the sole chronic prostatitis and warrants prostate biopsies to eliminate prostate cancer. Prostate 72:549–554, 2012. © 2011 Wiley Periodicals, Inc.

BACKGROUND The Androgen Receptor (AR) plays a key role in controlling prostate gland homeostasis and contributes to prostate carcinogenesis. The identification of its target genes should provide new candidates that may be implicated in cancer initiation and progression. METHODS Transcriptomic experiments and chromatin immunoprecipitation were combined to identify direct androgen regulated genes. Real-time quantitative PCR (RT-qPCR) analyses were performed to measure TM4SF1 mRNA levels in prostate cancer and benign prostatic hyperplasia (BPH) specimens. Immunohistochemical methods were used to compare TM4SF1 protein expression profiles in the same cohort. A targeted siRNAs knockdown strategy was used, prior to wound healing assays, to analyze the role of TM4SF1 in cell migration in vitro. RESULTS We demonstrate for the first time that TM4SF1 is a direct target gene of the AR, a transcription factor of the steroid nuclear receptor family. A functional androgen response element was identified in the promoter region of the gene. In addition, TM4SF1 mRNA expression was higher in cancer samples compared to BPH tissues. The TM4SF1 protein mediates cell motility of prostate cancer cells where it is predominantly localized in the cytoplasm, in contrast to its apical membrane localization in normal prostate epithelial cells. CONCLUSIONS Our results reveal a novel function for TM4SF1 in AR signaling. The TM4SF1 mRNA expression is higher in prostate cancer tissues as compared to BPH samples. Inhibition of cell migration after targeted knockdown of TM4SF1 protein expression suggests its contribution to prostate cancer cell metastasis. Prostate 71:1239–1250, 2011. © 2011 Wiley-Liss, Inc.

The prostate cancer 3 (PCA3) gene was discovered in 1999, on the basis of differential expression between cancer and noncancerous prostate tissue. Including the first study published in 2003, 11 clinical studies have evaluated its utility for the diagnosis of prostate cancer by measuring the number of PCA3 RNA copies in urine enriched with prostate cells. Although the sensitivity of the PCA3 test was less than that of serum prostate-specific antigen (PSA), its specificity appeared to be much better, particularly in patients with a previous negative biopsy. Recent studies also have suggested that this test could be used to predict cancer prognosis.

Resume Le gene PCA3 a ete decouvert en 1999 sur la base de son expression differentielle entre le cancer et le tissu prostatique non cancereux. Plusieurs etudes ont evalue l’interet diagnostique dans le cancer de la prostate de la mesure, dans les urines enrichies en cellules prostatiques, du nombre de copies des ARN produits par PCA3. Pour une sensibilite legerement inferieure a celle du dosage serique du PSA, la specificite et les valeurs predictives positive et negative de ce dosage (test PCA3) apparaissent meilleures. Le test PCA3 apparait ainsi comme un bon indicateur du resultat des biopsies prostatiques. La mise a disposition d’une trousse commerciale offre l’opportunite d’engager des etudes a grande echelle pour confirmer les resultats, preciser les indications du test et evaluer son interet en economie de la sante. L’un des objectifs est une meilleure selection des patients qui doivent etre orientes vers des biopsies prostatiques.

Management of patients with prostate cancer is currently based on imperfect clinical, biological, radiological and pathological evaluation. Prostate cancer aggressiveness, including metastatic potential, remains difficult to accurately estimate. In an attempt to better adapt therapeutics to an individual (personalized medicine), reliable evaluation of the intrinsic molecular biology of the tumor is warranted, and particularly for all tumor sites (primary tumors and secondary sites) at any time of the disease progression. As a consequence of their natural tendency to grow (passive invasion) or as a consequence of an active blood vessel invasion by metastase-initiating cells, tumors shed various materials into the bloodstream. Major efforts have been recently made to develop powerful and accurate methods able to detect, quantify and/or analyze all these circulating tumor materials: circulating tumors cells, disseminating tumor cells, extracellular vesicles (including exosomes), nucleic acids, etc. The aim of this review is to summarize current knowledge about these circulating tumor materials and their applications in translational research.

Context: RTH alpha is a recently discovered resistance to thyroid hormone (RTH) due to mutation of THRA, the gene encoding TR alpha 1, the thyroid hormone receptor. It has been described in a few patients with growth retardation, short stature, and a low free T-4/free T-3 (FT4/FT3) ratio. Objective: A 27-year-old patient presenting with dwarfism and a low FT4/FT3 ratio was investigated. Design: Clinical, biochemical, and radiological data were collected. Whole exome sequencing was performed in the patient and her relatives. Results: The patient exhibited congenital macrocytic anemia and severe bone malformation with growth retardation, dwarfism, clavicular agenesis, and abnormalities of the fingers, toes, and elbow joints. In adulthood, she presented with active behavior, chronic motor diarrhea, and hypercalcemia. Treatment with T3 led to heart rate acceleration, worsening of diarrhea, and TSH suppression. Low resting energy expenditure normalized on T-3. rT(3), SHBG, and IGF-1 remained normal. A de novo monoallelic missense mutation in THRA was discovered, the N359Y amino acid substitution (c. 1075A>T), which affected both the TR alpha 1 and the non-receptor isoform TR alpha 2. The mutant TR alpha 1 had a decrease in transcriptional activity related to decreased T-3 binding and a dominant-negative effect on the wild-type receptor. Conclusions: This patient presents a new phenotype including more significant bone abnormalities, lower TSH, and higher FT3 levels, without certainty of all her symptoms with the TR alpha 1(N359Y) mutation. This case suggests that patients with a low FT4/FT3 ratio should be screened for THRA mutations, even if clinical and biological features differ from previous reported cases of RTH alpha

International audience; Since 2012, eight different abnormalities have been described in the THRA gene (encoding the TRα1 thyroid hormone receptor) of 14 patients from 9 families. These mutations induce a clinical phenotype (resistance to thyroid hormone type α) associating symptoms of untreated mild congenital hypothyroidism and a near-normal range of free and total thyroid hormones and TSH (the T4/T3 ratio is nevertheless usually low). The phenotype can diversely include short stature (due to growth retardation), dysmorphic syndrome (face and limb extremities), psychoneuromotor disorders, constipation and bradycardia. The identified genetic abnormalities are located within the ligand-binding domain and result in defective T3 binding, an abnormally strong interaction with corepressors and a dominant negative activity against still functional receptors. The identification of patients with consistent phenotypes and the underlying mutations are warranted to better delineate the spectrum of the syndromes of reduced sensitivity to thyroid hormone.

Objectifs Le but de notre etude etait d’evaluer la capacite du test serique PHI a predire le resultat des biopsies prostatiques dans une large cohorte monocentrique. Methodes Entre 04/2010 et 05/2014, 594 patients adresses pour biopsies prostatiques du fait d’une suspicion de cancer prostatique (PSA eleve, TR anormal et/ou antecedent familial) ont ete inclus. Le PHI a ete determine a partir du dosage du PSA total, du PSA libre et du P2PSA (PHI = ([–2]proPSA/free PSA) × √(total PSA, Access Immunoassay Systems, Beckman Coulter ® ). Resultats Le PHI median etait significativement plus eleve chez les patients avec biopsies positives et Gleason ≥ 7. L’AUC etait respectivement de 0,79 et de 0,83. En analyse multivariee, le PHI etait predicteur independant du resultat des biopsies et son addition a un modele de base apportait un gain significatif. Au seuil de 25, 45 % des biopsies a posteriori inutiles auraient ete evitees tout en ayant ignore 4 % des cancers avec score de Gleason ≥ 7. Dans la meme population, le score PCA3 est apparu lui aussi comme un predicteur independant de la positivite des biopsies et l’association des deux marqueurs etait plus performante que chaque marqueur evalue individuellement ( Fig. 1 ). L’addition du PCA3 au PHI n’apportait par contre pas de gain supplementaire pour la prediction d’un cancer de Gleason ≥ 7 ( Fig. 2 ). Conclusion Le test serique PHI presente des performances diagnostiques elevees pour le diagnostic precoce du CaP. Son association au score urinaire PCA3 est utile pour predire la positivite des biopsies mais seul le PHI est utile pour predire un cancer de Gleason ≥ 7.

The efficacy of suppressing TSH secretion with levothyroxine (L-T(4)) in reducing solitary thyroid nodule growth is still controversial. In this prospective multicenter, randomized, double-blind, placebo-controlled trial, 123 patients with a single palpable benign nodule were included and randomly allocated to an 18-month treatment with L-T(4) or placebo. Individual dose was adjusted to allow a serum TSH level below 0.3 mIU/liter. Clinical and ultrasonographic nodule characteristics were assessed before treatment and 3, 6, 12, and 18 months thereafter. The largest mean nodule size assessed on palpation and largest volume, assessed by ultrasonography, decreased in the L-T(4) group and increased slightly in the placebo group [size, -3.5 +/- 7 mm vs. +0.5 +/- 6 mm (P = 0.006); volume, -0.36 +/- 1.71 ml vs. +0.62 +/- 3.67 ml (P = 0.01), respectively]. The proportion of clinically relevant volume reduction (or =50%) rose significantly in the L-T(4) group [26.6% vs. 16.9% (P = 0.04)]. The proportion of patients with a reduced number of infraclinical additional nodules was significantly higher in the L-T(4) group [9.4% vs. 0 (P = 0.04)]. It is concluded from this study that suppressive L-T(4) therapy is effective in reducing solitary thyroid nodule volume and improving infraclinical extranodular changes.

International audience; Objectives: Annexin A3 (ANXA3) is a potential marker for prostate cancer (PCa). We aimed to develop robust immunoassays suitable for quantifying ANXA3 in urine samples obtained following digital rectal examination (DRE) in order to facilitate the diagnostic performance evaluation of this marker. Design and methods: Anti-ANXA3 monoclonal antibodies were generated and their epitopes mapped. Two different ANXA3 assay prototypes were established on the VIDAS (R) automated immunoanalyser and analytical validation was carried out using post-DRE urine samples obtained from patients with PCa (n = 23) or benign prostate hyperplasia (n = 31). Results: The assays had the same capture antibody (TGC44) but different detection antibodies (13A12 or 5C5), recognizing novel distinct epitopes. Both had a lower limit of quantification

Objectifs Le but de notre etude etait d’evaluer la capacite du test urinaire PCA3 a predire le resultat des biopsies prostatiques dans une large cohorte monocentrique francaise. Methodes Entre 12/2007 et 05/2014, 1029 patients adresses pour biopsies prostatiques du fait d’une suspicion de cancer prostatique (PSA eleve, TR anormal et/ou antecedent familial) ont ete inclus. Le score urinaire PCA3 a ete determine a partir des mesures des nombres de copies des ARN de PCA3 et de PSA (Progensa PCA3®, DTS400 system, Hologic Gen-Probe). Resultats Le score PCA3 median etait significativement plus eleve chez les patients avec biopsies positives (47 %). L’AUC etait de 0,76, plus elevee que celle du PSA ( Fig. 1 ). Au seuil de 35, la sensibilite etait de 68 %, la specificite de 71 %, les VPP et VPN de 67 et 71 %. En analyse multivariee, le score PCA3 etait predicteur independant du resultat des biopsies et son addition a un modele de base comportant les donnees clinico-biologiques classiques apportait un gain diagnostique significatif (DCA : Fig. 2 ). Au seuil de 20, environ la moitie des biopsies, a posteriori inutiles, auraient ete evitees tout en ayant ignore 7 % des cancers avec score de Gleason ≥ 7. Le score PCA3 n’apparaissait pas correle au score de Gleason, mais etait bien correle au volume tumoral (proportion de carottes envahies). Conclusion Le test urinaire PCA3 est un test robuste avec des performances diagnostiques elevees pour le diagnostic precoce du CaP. Sa correlation avec l’agressivite du cancer s’exprime a travers le volume tumoral plus que par le score de Gleason.

Objectifs Il a ete suggere que l’index PHI pouvait etre utile pour le suivi de patients sous surveillance active pour cancer de la prostate [1] , [2] . Le but de cette etude etait de tester la relation entre l’index PHI a l’inclusion dans le protocole de surveillance active et le risque de reclassification biopsique. Methodes Notre etude, prospective, a inclus 64 patients en surveillance active sur les criteres suivants retrouves sur deux series successives de biopsies prostatiques realisees a 3 mois d’intervalle : T1c, densite de PSA Resultats Le p2PSA et le PHI medians etaient significativement plus eleves chez les 9 patients (14 %) avec cancer reclasse comme agressif a 12 mois. Seul PHI etait predictif independant de la reclassification. L’addition du PHI a un modele de base incluant l’âge et % PSA libre entrainait une augmentation de l’AUC de 0,76 a 0,84. Le suivi moyen des patients apres inclusion dans le protocole etait de 22 mois. A la date des dernieres nouvelles, 15 patients (23 %) avaient un cancer reclasse en agressif. L’âge a l’inclusion, le % PSA libre et PHI etaient predicteurs de la reclassification mais seul PHI etait independant. Un seuil de 33 pour PHI permettait le meilleur compromis entre sensibilite (67 %) et specificite (69 %). Les courbes de survie sans reclassification (modeles de Cox) etaient significativement differentes ( Fig. 1 ). Conclusion Le PHI realise au moment de l’inclusion dans un protocole de surveillance active permet de predire la reclassification des cancers indolents en cancers agressifs a 12 mois et au-dela.

While now recognized as an aid to predict repeat prostate biopsy outcome, the urinary PCA3 (prostate cancer gene 3) test has also been recently advocated to predict initial biopsy results. The objective is to evaluate the performance of the PCA3 test in predicting results of initial prostate biopsies and to determine whether its incorporation into specific nomograms reinforces its diagnostic value. A prospective study included 601 consecutive patients addressed for initial prostate biopsy. The PCA3 test was performed before ≥12-core initial prostate biopsy, along with standard risk factor assessment. Diagnostic performance of the PCA3 test was evaluated. The three available nomograms (Hansen’s and Chun’s nomograms, as well as the updated Prostate Cancer Prevention Trial risk calculator; PCPT) were applied to the cohort, and their predictive accuracies were assessed in terms of biopsy outcome: the presence of any prostate cancer (PCa) and high-grade prostate cancer (HGPCa). The PCA3 score provided significant predictive accuracy. While the PCPT risk calculator appeared less accurate; both Chun’s and Hansen’s nomograms provided good calibration and high net benefit on decision curve analyses. When applying nomogram-derived PCa probability thresholds ≤30%, ≤6% of HGPCa would have been missed, while avoiding up to 48% of unnecessary biopsies. The urinary PCA3 test and PCA3-incorporating nomograms can be considered as reliable tools to aid in the initial biopsy decision.

Prognosis of peritoneal surface malignancies is influenced by the adequacy of surgical and chemotherapeutic treatment and by tumor spread at the time of diagnosis. By promoting morphological changes in the mesothelium, inflammatory cytokines reflect tumor biology and could be evaluated as biomarkers. Our objective was to evaluate intraperitoneal levels of IL-6, IL-8, IL-10, TNF-alpha, and sICAM in patients with pseudomyxoma peritonei and peritoneal mesothelioma. Serum and peritoneal fluid samples were prospectively collected in patients managed for peritoneal surface malignancies including pseudomyxoma peritonei (PMP), mesotheliomas, and other rare primitive peritoneal cancers (cancer group) and patients who underwent intraperitoneal laparoscopic surgical procedures for benign diseases (noncancer group). Samples were analyzed for IL-6, IL-8, IL-10, TNF-alpha, and sICAM concentrations. Correlations were assessed with tumor spread related clinical scores. In both patient groups, intraperitoneal cytokine levels were higher than serum levels. Cancer patients had significantly higher intraperitoneal cytokine levels than noncancer patients. Peritoneal levels tended to increase in cancer patients with free tumor cells in peritoneal fluid. They were significantly higher in patients with tumor implants ≥2 cm and/or patients with peritoneal carcinomatosis index (PCI) >19. Furthermore, patients with malignant pseudomyxoma peritonei (grades II and III) had higher levels than patients with nonmalignant disease (grade I). Assessment of intraperitoneal IL-6, IL-8, IL-10, TNF-alpha, and sICAM levels can be performed in patients with peritoneal surface malignancies. They can be considered as both diagnostic and prognostic biomarkers that could be used as useful adjuncts for therapeutic decision making.

La faible specificite de la strategie diagnostique du cancer de la prostate (toucher rectal et dosage serique de l’antigene prostatique PSA) conduit a la realisation de nombreuses biopsies inutiles et au diagnostic de cancers indolents, sans potentialite evolutive. Un test urinaire (Progensa PCA3 ®, Gen-Probe) mesurant l’expression du gene PCA3, specifique des cellules cancereuses prostatiques, a recemment ete propose pour orienter les indications de re-biopsies. Le but de notre etude, prospective, etait d’evaluer l’interet diagnostique du score PCA3 dans le cancer de la prostate. Dans les urines de 245 patients adresses pour biopsie prostatique, l’expression du gene PCA3 a ete determinee par une technique d’amplification et de detection de l’ARN et rapportee a celle du PSA. Les patients avec echantillon informatif (98 %) ont ete classes selon la presence (n = 126) ou l’absence (n = 114) de cancer sur les biopsies. Le score PCA3 median etait plus eleve dans le groupe avec biopsies positives (p < 0,0001). L’aire sous la courbe ROC pour le PCA3 etait de 0,70 versus 0,53 pour le PSA total et 0,65 pour le rapport PSA libre/total. Au meilleur seuil de 38, le test PCA3 avait une sensibilite de 59 %, une specificite de 72 % (respectivement : 66 %, 32 % pour le PSA total au seuil de 4 ng/mL et 81 %, 28 % pour le rapport PSA libre/total au seuil de 25 %). Ces performances etaient maintenues chez les patients ayant un PSA dans la zone grise (4-10 ng/mL) ou ayant des antecedents de biopsies prostatiques. Notre etude confirme l’interet du test urinaire PCA3 dans l’aide a la decision de biopsies prostatiques.

The urinary PCA3 gene test has proved helpful for deciding whether to (re)biopsy to diagnose prostate cancer. We searched for pathological features that influence the shedding of PCA3 producing prostate cancer cells in urine after digital rectal examination.Included in our study were 102 patients with an informative PCA3 score on the Progensa® PCA3 assay who underwent radical prostatectomy. Correlations were evaluated between PCA3 score and histopathological factors on prostatectomy, including tumor site in the prostate and the number of cancer foci.PCA3 score significantly correlated with total tumor volume in prostatectomy specimens (p0.001) but not with prostatectomy Gleason score or pathological stage. PCA3 score positively correlated with apical and basal invasion, and with bilaterality and multifocality. On multivariate analysis multifocality was an independent factor influencing PCA3 score (p = 0.012).Site in the prostate gland and the number of cancer foci may explain the observed PCA3 score variation in patients operated on for prostate cancer. The PCA3 test could be helpful in preoperatively selecting patients with unifocal and unilateral cancer who could benefit from active surveillance or focal therapy.

Objectifs Certains cancers de la prostate sont associes a un test urinaire PCA3 faussement negatif. Leur proportion est variable, dependante du seuil utilise pour considerer le test comme positif ou negatif. L’objectif de cette etude etait de rechercher en quoi ces cancers avec PCA3 bas sont differents des cancers avec PCA3 eleve. Methodes Notre etude, monocentrique, a determine le score urinaire PCA3 (kit Progensa, Hologic–GenProbe ® ) chez 1015 patients adresses pour biopsies prostatiques du fait d’une suspicion de cancer prostatique. Ont ete consideres comme cancers avec PCA3 bas (CaP/PCA3 – ) les cancers associes a un score PCA3 Resultats Les patients avec un CaP/PCA3 – etaient plus jeunes (âge median : 62 vs 67 ; p p = 0,032) et un TR plus rarement suspect (8 vs 21 % ; p = 0,033). Le volume prostatique, la notion d’un antecedent familial de CaP ou de biopsies prostatiques negatives, la prise d-I5AR n’etaient pas des facteurs predictifs d’un score PCA3 faussement bas. Les cancers CaP/PCA3- etaient plus petits avec un nombre et une proportion de carottes envahies significativement plus bas (respectivement 2 vs 4 et 14 % vs 33 % ; p p p p = 0,005). Conclusion Les CaP avec un score PCA3 faussement negatif (

Objectifs Une correlation a ete demontree entre le score urinaire PCA3 et le volume tumoral sur piece de prostatectomie. Une correlation avec le caractere uni- ou bilateral et avec le nombre de foyers tumoraux (unifocalite anatomique) a egalement ete suggeree. Le but de cette etude etait d’evaluer la capacite du score PCA3 a predire l’unifocalite biologique et l’unilateralite. Methodes Une correlation a ete demontree entre le score urinaire PCA3 et le volume tumoral sur piece de prostatectomie. Une correlation avec le caractere uni- ou bilateral et avec le nombre de foyers tumoraux (unifocalite anatomique) a egalement ete suggeree. Le but de cette etude etait d’evaluer la capacite du score PCA3 a predire l’unifocalite biologique et l’unilateralite. Resultats Le score PCA3 median etait significativement plus bas chez les 41 patients avec tumeur anatomiquement unifocale (27 vs 51, p = 0,002). Il etait aussi plus bas chez les 60 patients avec tumeur biologiquement unifocale (40 vs 51, p = 0,005) et les 42 patients avec tumeur unilaterale (25 vs 54, p = 0,02). L’existence d’un envahissement des deux lobes prostatiques lors des biopsies etait un facteur predictif independant d’une atteinte bilaterale a la PR (AUC = 0,67). L’addition du score PCA3 a ce modele de regression a induit une augmentation significative de l’AUC a 0,73. Sur les 76 patients avec atteinte apparemment unilaterale sur les biopsies, 45 avaient finalement une atteinte bilaterale. Chez ces patients, le score PCA3 median etait significativement plus eleve que chez les 31 patients avec une atteinte unilaterale confirmee a la PR. Conclusion Le score PCA3 est un marqueur de l’unifocalite anatomique et biologique ainsi que du caractere uni- ou bilateral d’un CaP. Il pourrait constituer une aide pour le choix d’un traitement focal.

International audience; Amphibian metamorphosis is under the strict control of thyroid hormones (TH). These hormones induce metamorphosis by controlling gene expression through binding to thyroid hormone receptors (TRs). Necturus maculosus is considered to be an obligatory paedomorphic Amphibian since metamorphosis never occurs spontaneously and cannot be induced by pharmacological means. Since metamorphosis depends on the acquisition of response of tadpole tissues to thyroid hormone, we aimed to determine TR gene expression patterns in Necturus maculosusas well as the expression of two TH-related genes: Cytosolic Thyroid Hormone-Binding Protein (CTHBP)-M2-pyruvate kinase, a gene encoding a cytosolic TH binding protein and stromelysin 3, a direct TH target gene in Xenopus laevis. Tissue samples were obtained from specimens of Necturus maculosus. We performed in situ hybridization using non-cross-hybridizing RNA probes obtained from the cloned Necturus TR alpha and TR beta genes. We found clear expression of Necturus TRa gene in several tissues including the central nervous system, epithelial cells of digestive and urinary organs, as well as myocardium and skeletal muscle. TR beta was also expressed in the brain. In other tissues, hybridization signals were too low to draw reliable conclusions about their precise distribution. In addition, we observed that the expression of CTHBP and ST3 is largely distinct from that of TRs. The fact that we observed a clear expression of TR alpha and TR beta which are evolutionary conserved, suggests that Necturus tissues express TRs. Our results thus indicate that, in contrast to previously held hypotheses, Necturus tissues are TH responsive.

Several lines of evidence indicate that constraining intramolecular interactions between transmembrane domains are required to maintain G protein-coupled receptors in an inactive conformation in the absence of agonist. For the glycoprotein hormone receptors, which harbor a long amino-terminal ectodomain responsible for hormone binding, it has been suggested that the ectodomain could contribute to these negative constraints. To test this hypothesis, we expressed at the surface of COS-7 cells mutants of the TSH receptor in which variable portions of the amino-terminal ectodomain are replaced by a 19-residue tag from bovine rhodopsin. Whereas none of the rhodopsin-tagged truncated mutants could be activated by saturating concentrations of TSH, the constructs with the shortest amino-terminal extension displayed increased constitutive activity toward the cAMP pathway, when compared with the wild-type holoreceptor. The shortest truncated construct was strongly activated by the introduction of mutations in transmembrane segment VI (D633A), or in the third intracellular loop (A623I) of the receptor. The magnitude of the stimulation was similar to that observed when the same mutations were introduced in the intact wild-type receptor. On the contrary, the shortest truncated construct was unaffected by activating mutations affecting residues of the extracellular loop region (I486F, I568T) or the top of transmembrane segment VII (del658-661). Together, our results are compatible with a model in which activation of the cAMP pathway by the TSH receptor involves switching of the ectodomain from a tethered inverse agonist to a true agonist.

Pseudohypoparathyroidism Ia (PHP Ia) is characterized by resistance to PTH and many other stimuli because of deficiency of stimulatory G protein alpha-subunit. To determine the incidence, natural history, and mechanism of C cell dysfunction in PHP, calcitonin assays were performed in six patients with PHP Ia and four with pseudopseudohypoparathyroidism from three unrelated families. Controls included healthy subjects and patients with PHP Ib or hypoparathyroidism. The mean basal level of calcitonin was higher in PHP Ia patients than in controls (95.3 +/- 112.7 vs. 3.7 +/- 2.4 pg/mL; P = 0.005; n10). In PHP Ia patients, calcitonin levels rose over the normal range (30 pg/mL) after pentagastrin infusion in five patients and remained normal in one. Familial medullary thyroid carcinoma was clinically, biologically, and ultrasonographically ruled out over a mean follow-up exceeding 3 yr. Genomic screening for RET protooncogene mutations failed to reveal any anomaly. The calcitonin infusion test, which induced a significant increase in plasma cAMP in controls 30 and 60 min after infusion, failed to produce this response in PHP Ia patients, suggesting that the action of calcitonin was specifically impaired. PHP Ia may therefore be an independent etiology of hypercalcitoninemia and hyperresponsiveness to pentagastrin infusion.

International audience; Background: Several data favor androgen receptor implication in prostate cancer initiation through the induction of several gene activation programs. The aim of the study is to identify potential biomarkers for early diagnosis of prostate cancer (PCa) among androgen-regulated genes (ARG) and to evaluate comparative expression of these genes in normal prostate and normal prostate-related androgen-sensitive tissues that do not (or rarely) give rise to cancer. Methods: ARG were selected in non-neoplastic adult human prostatic epithelial RWPE-1 cells stably expressing an exogenous human androgen receptor, using RNA-microarrays and validation by qRT-PCR. Expression of 48 preselected genes was quantified in tissue samples (seminal vesicles, prostate transitional zones and prostate cancers, benign prostatic hypertrophy obtained from surgical specimens) using TaqManH low-density arrays. The diagnostic performances of these potential biomarkers were compared to that of genes known to be associated with PCa (i.e. PCA3 and DLX1). Results and Discussion: By crossing expression studies in 26 matched PCa and normal prostate transitional zone samples, and 35 matched seminal vesicle and PCa samples, 14 genes were identified. Similarly, 9 genes were overexpressed in 15 benign prostatic hypertrophy samples, as compared to PCa samples. Overall, we selected 8 genes of interest to evaluate their diagnostic performances in comparison with that of PCA3 and DLX1. Among them, 3 genes: CRYAB, KCNMA1 and SDPR, were overexpressed in all 3 reference non-cancerous tissues. The areas under ROC curves of these genes reached those of PCA3 (0.91) and DLX1 (0.94). Conclusions: We identified ARG with reduced expression in PCa and with significant diagnostic values for discriminating between cancerous and non-cancerous prostatic tissues, similar that of PCA3. Given their expression pattern, they could be considered as potentially protective against prostate cancer. Moreover, they could be complementary to known genes overexpressed in PCa and included along with them in multiplex diagnostic tools.

International audience; Prostate-specific antigen (PSA) is a serum marker that is widely used for the diagnosis of prostatic diseases. Various subforms of free PSA, which are associated with prostate cancer differently, have been identified in sera. Thus, specific detection of certain subforms could permit discrimination between benign and malignant cases. Although the monoclonal antibody 5D3D11 displays the desired selectivity, its relative weak binding affinity prevents its development into an effective diagnostic tool. The directed-evolution strategy presented here succeeds in enhancing affinity and immunoassay sensitivity while maintaining selectivity. Starting without structural data, we constructed four independent phage-display single-chain variable fragment (scFv) libraries targeting hot spots from CDR-L1, H1, H2, and H3. Mutations derived from each library were combined, yielding further affinity gains. This constitutes the first demonstration of additivity for independently selected complementarity-determining region (CDR) hot-spot mutations. The X-ray structure of the Fab′ 5D3D11–PSA complex (after it became available) inspired the design of two new libraries targeting CDR-L3 that resulted in other higher-affinity variants. Attempts at combining the new variants with previous ones did not result in further gains, suggesting that mutations from the two strategies provide alternative but noncomplementary solutions for affinity enhancement of 5D3D11. The results can be interpreted to provide a plausible explanation for the observed lack of additivity. Finally, with respect to the wild-type scFv, the best binders show an enhancement of sensitivity in sandwich immunoassay. Its ability to discriminate between prostate cancer sera and benign prostatic hyperplasia sera has now been confirmed through the dosage of 63 patients.