First evidence of subclinical renal tubular injury during sickle-cell crisis

International audienceBACKGROUND: The pathophysiologic mechanisms classically involved in sickle-cell nephropathy include endothelial dysfunction and vascular occlusion. Arguments demonstrating that ischemia-reperfusion injury-related kidney damage might coincide with vaso-occlusive crisis (VOC) are lacking. METHODS: In this prospective study, we sought to determine whether tubular cells and glomerular permeability might be altered during VOC. Urine neutrophil gelatinase-associated lipocalin (NGAL) levels and albumin-excretion rates (AER) of 25 patients were evaluated prospectively during 25 VOC episodes and compared to their steady state (ST) values. RESULTS: During VOC, white blood-cell counts (WBC) and C-reactive protein (CRP) were significantly higher than at ST but creatinine levels were comparable. Urine NGAL levels were significantly increased during VOC vs ST (P = 0.007) and remained significant when normalized to urine creatinine (P = 0.004), while AER did not change significantly. The higher urine NGAL concentration was not associated with subsequent (24-48 hour) acute kidney injury. Univariate analysis identified no significant correlations between urine NGAL levels and laboratory parameters during VOC. CONCLUSIONS: These results demonstrated that subclinical ischemia-reperfusion tubular injury is common during VOC and highlight the importance of hydroelectrolyte monitoring and correction during VOC

Métabolisme oxydatif et mitochondrial des cellules cancéreuses de prostate

Pas de résumé françaisPas de résumé anglaisPARIS-EST-Université (770839901) / SudocSudocFranceF

Potentiel métastatique des cellules circulantes dans le cancer colorectal

Les cellules malignes circulantes auraient un rôle majeur dans le processus métastatique des tumeurs épithéliales. Le but de ce travail a été d en étudier le rôle pronostique dans le cancer colorectal à travers le développement de plusieurs techniques moléculaires de détection. La détection par RT-PCR CGM2 dans le sang périphérique préopératoire n est corrélée ni à la survie ni à la survie sans récidive. La détection par RT-PCR à l aide de deux marqueurs spécifiques du tissu colique (ACE/CGM2) permet soit d augmenter la spécificite de la détection soit d appréhender l existence de plusieurs sous-populations de cellules circulantes. L intérêt des techniques de RT-PCR quantitatives a été évalué pour CK20. La dissémination supplémentaire liée à l exérèse des cancers colorectaux a été évaluée à 10% environ par une technique de RT-PCR qualitative multiplex (ACE/CGM2). Ces résultats ont été comparés à ceux obtenus avec des marqueurs spécifiques de cancer (MAGE). La valeur pronostique des cellules malignes circulantes dans les cancers du colon et de la prostate. L intérêt des cellules circulantes dans la cavité péritonéale a été discuté. Une technique d isolement cellulaire a permis de récupérer plus de 98% des cellules nucléées sanguines, devenant ainsi la première étape de l étude physique du patrimoine génétique et du potentiel métastatique des cellules circulantes.Circulating cancer cells have been thought to have a great part in the development of distant metastasis in epithelial tumours. The aim of this work was to assess their prognostic value in colorectal cancer using several detection techniques. RT-PCR CGM2 detection in the preoperative peripheral blood is not correlated with either survival or recurrence-free survival. RT-PCR detection using two specific markers of the colon tissue (ACE/CGM2) either allows to improve detection specificity or to label several subpopulations of circulating cells. The interest of quantitative RT-PCR techniques was assessed for CK20. The additional dissemination related to colorectal cancer excision has been assessed to be about 10% using a qualitative multiplex (ACE/CGM2) RT-PCR technique. These results were compared to those obtained with cancer-specific markers (MAGE). The prognostic value of circulating malignant cells has been compared in colorectal and prostate cancers. The potential interest of circulating cells in peritoneal cavity has also been studied. An isolation technique has allowed to retrieve more than 98% of blood nucleated cells and thus become the first step to the physical study of the genotype and metastatic potential of circulating cells.PARIS12-CRETEIL BU Multidisc. (940282102) / SudocSudocFranceF

Implication de PAR1 dans la progression du cancer de la prostate

Le phénomène métastatique est important à comprendre puisque de manière irrémédiable, une fois engagé, il conduit le plus souvent au décès des patients. Le cancer de la prostate représente un bon modèle car sa progression du stade hormono-dépendant vers le stade d'hormono-échappement s accompagne par l apparition de métastases. Les Protease Activated Receptors (PAR1-4) sont des récepteurs qui jouent un rôle important dans l'hémostase et l'inflammation et dont l implication dans la prolifération et l'invasion des cellules tumorales a été décrite dans plusieurs tissus. L étude comparative de l'expression in vitro de PAR1 a confirmé son rôle dans la prolifération et l'invasion des lignées prostatiques normales et tumorales hormono-sensibles comme hormono-indépendantes. Son expression in vivo dans des tissus prostatiques à différents stades pathologiques a montré une surexpression de PAR1 chez les patients ayant atteint le stade d'hormono-échappement, associée à un mauvais pronostic. Son absence s avère par contre de bon pronostic chez les patients hormonodépendants. L'ensemble des résultats obtenus nous permet de proposer PAR1 comme un nouveau marqueur pronostique pour le cancer de la prostate. L activation des PARs, comme celle de plusieurs récepteurs de chémokines, apparaît comme un élément fondateur de la transition vers l état métastatique. Le décryptage de cette combinatoire permettra de mieux comprendre les phénomènes impliqués dans cette transition et permettra de développer des thérapies ciblées pour prévenir l apparition délétère de métastasesMetastasis is nowadays an important field of research as, once engaged, patients will generally die from their metastatic cancer. Prostate cancer represents an interesting model as its progression from hormone-naïve to hormone-independent status lead to metastatic disease. Protease Activated Receptors (PAR1-4) are G-protein-coupled receptors that play crucial roles in blood coagulation and inflammation but that are likely to play fundamental role in tumor cells proliferation and invasion. In vitro analysis of PAR1 expression in prostate cancer cell lines has confirmed the role of PAR1 in prostate cancer proliferation and invasion. Its expression in vivo in prostate cancer tissues have shown a constant surexpression in hormonerefractory ones, associated with a worse prognosis. However, its absence in hormone-naïve tissues is associated with a good prognosis. These results prompted us to recommend PAR1 as a new prognostic marker associated with prostate cancer progression. PAR activation, as well as several chemokine receptors, seems to be a founder feature of cancer transition to metastasis. Deciphering the pattern of receptor activation will allow a better understanding of the events that drive transition to metastasis and thus the development of new specific targeted therapeutics aimed at stopping deleterious metastatic evolutionPARIS-EST-Université (770839901) / SudocSudocFranceF

Prognostic value of circulating prostate cells in patients with a rising PSA after radical prostatectomy.

BACKGROUND: To predict poor outcome in patients with a biochemical recurrence (rising PSA) after radical prostatectomy (RP), urologists rely primarily on Gleason score, PSA doubling time, and time from surgery to biochemical (i.e., PSA) recurrence. In the present study, we assess the value of RT-PCR detection circulating prostate cells in blood of patients with a rising PSA. METHODS: RNA from blood samples was obtained from 55 patients with a rising PSA and from 45 patients without evidence of biochemical failure (PSA < 0.1 ng/ml). Both groups were matched for age, Gleason score, pT stage, and interval between radical prostatectomy and PCR testing. RESULTS: PSA positive cells were detected in 1/45 (2%) patients without a PSA recurrence and 19/55 (34%) patients with a PSA recurrence. In the rising PSA group, mean PSA doubling time was significantly shorter in patients with positive RT-PCR (5 months) than in patients with negative RT-PCR (16 months; P = 0.001). An earlier onset of recurrence was also detected in patients with a positive RT-PCR (31 months for positive RT-PCR vs. 50 months for negative RT-PCR) but this result did not achieve statistical significance (P = 0.102). Salvage radiation therapy was administered in 15 patients. Three of the five patients with a positive RT-PCR progressed during radiotherapy whereas 7 of the 10 patients with a negative RT-PCR obtained a complete response and none have progressed. CONCLUSIONS: These preliminary results suggest that RT-PCR detection of prostate cells in blood of patients after RP correlates with rapidly progressing biochemical failure after RP

Functional serotonin-2B receptors are expressed by a teratocarcinoma-derived cell line during serotoninergic differentiation.

International audienceAmong immortalized teratocarcinoma-derived cells, the clone 1C11 is a committed precursor of the neuronal lineage. On day 2 of its serotoninergic differentiation, this clone expresses only one subtype of serotonin [5-hydroxytryptamine (5-HT)] receptor, which is functionally coupled to phosphatidylinositol hydrolysis. The identity of these receptors was established by comparing their properties with those of 5-HT2B receptors expressed by LMTK- fibroblasts stably transfected with the recently cloned murine cDNA NP75 (LM5 cells). In both cell types, the analysis of (+/-)-1-(2,5-dimethoxy-4-[125I]iodophenyl)- 2-aminopropane HCl ([125I]DOI) binding revealed the presence of a single class of sites, the affinity of which was 1 order of magnitude lower than that reported for 5-HT2A receptors. In 1C11 cells differentiated for 2 days, as well as in LM5 cells, DOI binding was decreased by nonhydrolyzable analogs of GTP, indicating that the 5-HT2B receptor is functionally coupled to a G protein. The DOI-induced increase of phosphoinositide hydrolysis, which was correlated with both GTPase activity and binding data, is mediated by a Gq protein. This work demonstrates that the 5-HT2B receptor is functionally expressed before complete serotoninergic differentiation of 1C11 cells. The inducible 1C11 clone thus provides an in vitro model to investigate the possible role of the 5-HT2B receptor in the expression of the serotoninergic phenotype

Functional serotonin-2B receptors are expressed by a teratocarcinoma-derived cell line during serotoninergic differentiation.

International audienceAmong immortalized teratocarcinoma-derived cells, the clone 1C11 is a committed precursor of the neuronal lineage. On day 2 of its serotoninergic differentiation, this clone expresses only one subtype of serotonin [5-hydroxytryptamine (5-HT)] receptor, which is functionally coupled to phosphatidylinositol hydrolysis. The identity of these receptors was established by comparing their properties with those of 5-HT2B receptors expressed by LMTK- fibroblasts stably transfected with the recently cloned murine cDNA NP75 (LM5 cells). In both cell types, the analysis of (+/-)-1-(2,5-dimethoxy-4-[125I]iodophenyl)- 2-aminopropane HCl ([125I]DOI) binding revealed the presence of a single class of sites, the affinity of which was 1 order of magnitude lower than that reported for 5-HT2A receptors. In 1C11 cells differentiated for 2 days, as well as in LM5 cells, DOI binding was decreased by nonhydrolyzable analogs of GTP, indicating that the 5-HT2B receptor is functionally coupled to a G protein. The DOI-induced increase of phosphoinositide hydrolysis, which was correlated with both GTPase activity and binding data, is mediated by a Gq protein. This work demonstrates that the 5-HT2B receptor is functionally expressed before complete serotoninergic differentiation of 1C11 cells. The inducible 1C11 clone thus provides an in vitro model to investigate the possible role of the 5-HT2B receptor in the expression of the serotoninergic phenotype

Extracellular vesicles: General features and usefulness in diagnosis and therapeutic management of colorectal cancer

International audienceIn the world, among all type of cancers, colorectal cancer (CRC) is the third most commonly diagnosed in males and the second in females. In most of cases, (RP1) patients’ prognosis limitation with malignant tumors can be attributed to delayed diagnosis of the disease. Identification of patients with early-stage disease leads to more effective therapeutic interventions. Therefore, new screening methods and further innovative treatment approaches are mandatory as they may lead to an increase in progression-free and overall survival rates. For the last decade, the interest in extracellular vesicles (EVs) research has exponentially increased as EVs generation appears to be a universal feature of every cell that is strongly involved in many mechanisms of cell-cell communication either in physiological or pathological situations. EVs can cargo biomolecules, such as lipids, proteins, nucleic acids and generate transmission signal through the intercellular transfer of their content. By this mechanism, tumor cells can recruit and modify the adjacent and systemic microenvironment to support further invasion and dissemination. This review intends to cover the most recent literature on the role of EVs production in colorectal normal and cancer tissues. Specific attention is paid to the use of EVs for early CRC diagnosis, follow-up, and prognosis as EVs have come into the spotlight of research as a high potential source of ‘liquid biopsies’. The use of EVs as new targets or nanovectors as drug delivery systems for CRC therapy is also summarized

Extracellular Vesicles in Breast Cancer: From Biology and Function to Clinical Diagnosis and Therapeutic Management

Breast cancer (BC) is the first worldwide most frequent cancer in both sexes and the most commonly diagnosed in females. Although BC mortality has been thoroughly declining over the past decades, there are still considerable differences between women diagnosed with early BC and when metastatic BC is diagnosed. BC treatment choice is widely dependent on precise histological and molecular characterization. However, recurrence or distant metastasis still occurs even with the most recent efficient therapies. Thus, a better understanding of the different factors underlying tumor escape is mainly mandatory. Among the leading candidates is the continuous interplay between tumor cells and their microenvironment, where extracellular vesicles play a significant role. Among extracellular vesicles, smaller ones, also called exosomes, can carry biomolecules, such as lipids, proteins, and nucleic acids, and generate signal transmission through an intercellular transfer of their content. This mechanism allows tumor cells to recruit and modify the adjacent and systemic microenvironment to support further invasion and dissemination. By reciprocity, stromal cells can also use exosomes to profoundly modify tumor cell behavior. This review intends to cover the most recent literature on the role of extracellular vesicle production in normal and cancerous breast tissues. Specific attention is paid to the use of extracellular vesicles for early BC diagnosis, follow-up, and prognosis because exosomes are actually under the spotlight of researchers as a high-potential source of liquid biopsies. Extracellular vesicles in BC treatment as new targets for therapy or efficient nanovectors to drive drug delivery are also summarized