c-Jun-N-Terminal Kinase Signaling Is Involved in Cyclosporine-Induced Epithelial Phenotypic Changes

Tubular epithelial cells play a central role in the pathogenesis of chronic nephropathies. Previous toxicogenomic studies have demonstrated that cyclosporine- (CsA-) induced epithelial phenotypic changes (EPCs) are reminiscent of an incomplete epithelial to mesenchymal transition (EMT) in a TGF-β-independent manner. Furthermore, we identified endoplasmic reticulum (ER) stress as a potential mechanism that may participate in the modulation of tubular cell plasticity during CsA exposure. Because c-jun-N-terminal kinase (JNK), which is activated during ER stress, is implicated in kidney fibrogenesis, we undertook the current study to identify the role of JNK signaling in EPCs induced by CsA. In primary cultures of human renal epithelial cells, CsA activates JNK signaling, and the treatment with a JNK inhibitor reduces the occurrence of cell shape changes, E-cadherin downregulation, cell migration, and Snail-1 expression. Our results suggest that CsA activates JNK signaling, which, in turn, may participate in the morphological alterations through the regulation of Snail-1 expression

Conséquences rénales de l'activation de la réponse UPR (Unfolded protein response) par des stress toxique et ischémique

Le rein natif et le greffon rénal peuvent être soumis à de multiples agressions conduisant à la détérioration progressive du parenchyme. Ces agressions peuvent être spécifiques (stress toxique, immunologique) et/ou non spécifiques (stress ischémique) et vont engendrer des réponses pouvant entraîner à la fois une diminution de la consommation d énergie, une augmentation des apports afin de maintenir l homéostasie tissulaire et la survie mais aussi une réaction inflammatoire et l apoptose pouvant conduire à la fibrose. Parmi celles-ci, on peut nommer les voies HIF1a, mTOR, le stress du réticulum endoplasmique (RE), l autophagie, l activation de l immunité innée et acquise. La réponse adaptative qui suit le stress du RE, la réponse UPR (Unfolded protein response), est une voie adaptative dont les implications sont actuellement encore peu connues dans le domaine de la pathologie rénale. Celle-ci se compose de trois effecteurs principaux : Perk, Ire1 et ATF6. A l aide de deux modèles de stress toxique (ciclosporine) et ischémique (carence en glucose) sur deux modèles cellulaires distincts (cellulaires endothéliales artérielles et cellules tubulaires rénales), et dans des modèles in vivo, nous avons montré que le stress du RE était impliqué à la fois dans l apparition de modifications phénotypiques endothéliales évocatrices de transition endothélio-mésenchymateuse induites par la ciclosporine et à la fois dans l induction de réponses inflammatoire (régulation de NF- B par Ire1) et angiogénique (régulation distincte de VEGF, bFGF et angiogénine par Perk et Ire1) induites par la carence en glucose. La réponse UPR semble modulée de façon subtile au cours de ces stress car les trois effecteurs n engendrent pas des réponses identiques. Ces travaux apportent ainsi une meilleure compréhension des mécanismes d adaptation au cours de stress variés, montrent que le stress du RE est impliqué dans ces réponses adaptatives et que la réponse peut être différente selon les effecteurs de la réponse UPR. Cette meilleure compréhension pourra permettre de valider des biomarqueurs précoces et des modulateurs de la réponse UPR afin de prévenir la dégradation du parenchyme rénal.Native and grafted kidneys are stressed by multiple specific or non-specific insults leading to progressive structural deterioration. Responses to these insults are adaptive and preserve cell survival but may also promote inflammation, fibrosis and apoptosis. The most important of these adaptive pathways are HIF1a pathway, mTOR pathway, autophagy, unfolded protein response (UPR). The consequences of the UPR in kidney injuries are not well known. The objective of this study is to delineate the mechanisms and consequences of the activation of the UPR in response to toxic (cyclosporine) and ischemic (glucose starvation) stresses in two distinct cellular models (arterial endothelial cells and renal tubular cells). Here, we showed that UPR was engaged in cyclosporine-induced endothelial phenotypic changes, glucose starvation-induced inflammatory and angiogenic responses: NF- B regulation by Ire1; distinct VEGF, bFGF and angiogenin regulation by Perk and Ire1. UPR is subtly modulated since its transducers do not induce identical processes. In conclusion these comprehensive works, we demonstrate the UPR is implicated in stress-induced adaptive pathways with different downstream responses according to the effector. Renal tissue degradation could be prevented by discovering and validating early biomarker and UPR modulators.PARIS5-Bibliotheque electronique (751069902) / SudocPARIS-BIUM-Bib. électronique (751069903) / SudocSudocFranceF

Rôle du stress du réticulum endoplasmique et de l'autophagie dans la régulation des réponses immune et angiogénique activées par des stress ischémiques et inflammatoires dans l'épithélium rénal humain

Dans le cadre de situations pathologiques, le rein peut être soumis à de multiples agressions toxiques, ischémiques et immunologiques pouvant favoriser la survenue d une maladie rénale chronique et le développement d une insuffisance rénale. En réponse à ces stress, les cellules du parenchyme rénal vont activer des processus biologiques adaptatifs permettant le maintien de la viabilité cellulaire et l homéostasie de l organe. Ces réponses adaptatives peuvent également activer l immunité innée et induire le remodelage tissulaire (fibrogenèse et angiogenèse). Cependant, les mécanismes précis de cette régulation sont mal connus. L objectif de ce travail a été de caractériser les mécanismes de régulation et les conséquences microenvironnementales (inflammation et angiogenèse) de l activation de la réponse UPR (Unfolded Protein Response) et de l autophagie, en réponse à des stress ischémiques et immunologiques. Dans un premier travail, nous avons montré que la réponse UPR est impliquée dans la génération d une réponse inflammatoire induite par un stress métabolique dans des cellules tubulaires rénales. Le stress métabolique, caractérisé par une carence en glucose, induit un stress du RE et active la réponse UPR. Ce stress active le facteur NF-.B et favorise la transcription de cytokines et chimiokines pro-inflammatoires. La voie PERK/eIF2 : - n est pas nécessaire à l activation de l inflammation mais amplifie l expression des cytokines alors que la voie IRE1 - est impliquée dans la génération de cette réponse inflammatoire. De plus, l ischémie aigue active le stress du RE et l inflammation dans les reins de rat. Enfin, à partir de biopsies de déclampage de greffons rénaux, l expression de GRP78, marqueur du stress du RE, et de NF-.B p65/RelA dans les tubules rénaux, est significativement plus élevée en comparaison avec des biopsies de greffons rénaux stables, à distance de la greffe. Dans un second travail, nous avons montré que la réponse UPR régule l angiogenèse dans les cellules tubulaires rénales lors d une carence en glucose. La voie PERK est un régulateur majeur de l expression des facteurs angiogéniques (VEGFA, bFGF et angiogénine). De plus, l expression de l angiogénine est modulée par les voies PERK et IRE1.. Enfin, l ischémie aigue induite chez le rat, active la réponse UPR parallèlement à l augmentation de l expression de VEGFA, bFGF et de l angiogénine. Dans un troisième travail, nous avons mis en évidence un nouveau mécanisme par lequel l interféron. (IFN.) active l autophagie dans les cellules tubulaires rénales. Nous avons montré que l IFN. entraine une déplétion en tryptophane, active la voie GCN2, une kinase eIF2., ce qui conduit à l augmentation du flux autophagique. De plus, la supplémentation entryptophane et l utilisation d ARN interférence dirigés contre GCN2 inhibent l autophagie induite par l IFN. Enfin, l autophagie intervient dans la régulation de la sécrétion de cytokines inflammatoires et de facteurs de croissance en réponse à l IFN.. En conclusion, nous avons caractérisé dans ce travail des mécanismes originaux de régulation d une réponse inflammatoire et angiogénique par la réponse UPR et l autophagie en réponse à des stress ischémiques et immunologiques au sein de l épithélium tubulaire rénal humain.Under pathological conditions, kidney is subjected to multiple toxic, ischemic and immunological failures that promote the occurrence of chronic kidney disease and the development of acute kidney injury. In response to stress, renal parenchymal cells activate biological adaptive processes permitting the maintenance of cell viability and renal homeostasis. These adaptive responses can also activate innate immunity and induce tissue remodeling (fibrogenesis and angiogenesis). However, accurate mechanisms of this regulation are still unclear. The aim of this work was to characterize regulation mechanisms and micro environmental consequences(inflammation and angiogenesis) of the activation of the UPR (Unfolded Protein Response) and autophagy, in response to ischemic and immunological stress. In a first study, we demonstrated that the UPR is involved in the generation of inflammatory response induces by metabolic stress in tubular renal cells. Metabolic stress, characterized by glucose deprivation, induces an ER stress and activates the UPR. This stress activates NF-.B and promotes the transcription of pro inflammatory cytokines and chemokines. The PERK signaling is not required for the induction of inflammation but amplifies cytokine expression whereas IRE1 is involved in the generation of inflammatory response. Moreover, acute ischemia activates ER stress and inflammation in rat kidneys. Finally, from kidney transplant biopsies performed before implantation, the expression of GRP78, an ER stress marker, and NF-.B p65/RelA in renal tubules is significantly increased in comparison with stable human kidney transplant biopsies. In a second study, we showed that the UPR regulates angiogenesis in tubular renal cells during glucose deprivation. The PERK pathway is a major regulator of angiogenic factors expression (VEGFA, bFGF and angiogenin). Furthermore, angiogenin expression is modulated by PERK and IRE1. pathways. Finally, acute ischemia activates the UPR and, in parallel, increases VEGFA, bFGF and angiogenin expression in rat kidneys. In a third work, we identified a novel mechanism by which IFN. activates autophagy in human kidney epithelial cells. We showed that IFN. promotes tryptophan depletion, activates the eIF2. kinase GCN2, and leads to an increase of the autophagic flux. Moreover, tryptophan supplementation and RNA interference directed against GCN2 inhibit IFN.-induced autophagy. Finally,autophagy regulates the secretion of inflammatory cytokines and growth factors in response to IFN..In conclusion, we characterized in this work original mechanisms that regulate inflammatory and angiogenic responses by the UPR and autophagy in response to ischemic and immunological stress in tubular renal human epithelium.PARIS5-Bibliotheque electronique (751069902) / SudocPARIS-BIUM-Bib. électronique (751069903) / SudocSudocFranceF

Home and Office Blood Pressure Monitoring in Renal Transplant Recipients

Background. Arterial hypertension in renal transplant recipients (RTR) is associated with increased morbid mortality. In the general population, home blood pressure monitoring (HBPM) was found to be superior to office blood pressure (OBP) in identifying true hypertensive patients. The aim of this study was to investigate HBPM for the assessment of blood pressure profile in RTR. Methodology and Principal Findings. We included prospectively 87 stable RTR. Sitting OBP was measured during the outpatient clinic. HBPM was performed by measuring BP every morning and night for 4 days. The accepted limits for the OBP and HBPM, were respectively, 140/90 mmHg and 135/85 mmHg. Patients were classified as “normotensive,” “uncontrolled,” “white-coat hypertensive” and “masked hypertensive”, (OBP below the limit and HBPM above). During the study, 81 patients (55 males, age 48.5 ± 14 years) were available for analysis. The mean OBP and HBP were 138/83 ± 14/10 mmHg and 133/79 ± 14/8 mmHg; 29% of patients were uncontrolled, 28% normotensive, 21% white coat, and 21% masked hypertensive. Age, glycemia, and number of antihypertensive drugs were associated with hypertension. Conclusion and Significance. In RTR, HBPM is well accepted and better define BP profile since there is 42% discrepancy between OBPM and HBPM. Whether this discrepancy is associated with worst outcome in the long term remains to be demonstrated

Special RepoRt Practical recommendations for pharmacogenomics- based prescription: 2010 ESF-UB Conference on Pharmacogenetics and Pharmacogenomics

medical practice Oncology drugs A full day was dedicated to oncology covering germline as well as tumor pharmacogenomics. Three major examples were discussed. Response to tyrosine kinase inhibitors owing to activating EGFR mutations in non-small-cell lung cancer Miguel A Molina from Instituto Universitario USP Dexeus, Barcelona, presented the results of a national survey indicating the usefulness of tumor EGFR pharmacogenomics in order to define tumors that will respond (owing to activating mutations) to EGF receptor (EGFR) antagonists (tyrosine kinase inhibitors) [1]. Additional recent publications have confirmed the usefulness of EGFR pharmacogenomics in non-small-cell lung cancer (NSCLC) [2,3]. Tumor samples can be obtained from tumor biopsies, possibly followed by laser microdissection -or circulating blood tumor cells. Activating mutations are observed in 15% of The present article summarizes the discussions of the 3rd European Science Foundation-University of Barcelona (ESF-UB) Conference in Biomedicine on Pharmacogenetics and Pharmacogenomics, which was held in June 2010 in Spain. It was focused on practical applications in routine medical practice. We provide practical recommendations for ten different clinical situations, that have either been approved or not approved by regulatory agencies. We propose some comments that might accompany the results of these tests, indicating the best drug and doses to be prescribed. The discussed examples include KRAS, cetuximab, panitumumab, EGFR-gefitinib, CYP2D6 -tamoxifen, TPMT-azathioprine -6-mercaptopurine, VKORC1/CYP2C9-warfarin, CYP2C19-clopidogrel, HLA-B*5701-abacavir, HLA-B*5701-flucloxacillin, SLCO1B1-statins and CYP3A5-tacrolimus. We hope that these practical recommendations will help physicians, biologists, scientists and other healthcare professionals to prescribe, perform and interpret these genetic tests. KEYWORDS: adverse drug reaction azathioprine cetuximab clopidogrel gefitinib genetic testing pharmacogenetics statins tacrolimus tamoxifen warfari

Canagliflozin and Cardiovascular and Renal Outcomes in Type 2 Diabetes Mellitus and Chronic Kidney Disease in Primary and Secondary Cardiovascular Prevention Groups

Background: Canagliflozin reduces the risk of kidney failure in patients with type 2 diabetes mellitus and chronic kidney disease, but effects on specific cardiovascular outcomes are uncertain, as are effects in people without previous cardiovascular disease (primary prevention). Methods: In CREDENCE (Canagliflozin and Renal Events in Diabetes With Established Nephropathy Clinical Evaluation), 4401 participants with type 2 diabetes mellitus and chronic kidney disease were randomly assigned to canagliflozin or placebo on a background of optimized standard of care. Results: Primary prevention participants (n=2181, 49.6%) were younger (61 versus 65 years), were more often female (37% versus 31%), and had shorter duration of diabetes mellitus (15 years versus 16 years) compared with secondary prevention participants (n=2220, 50.4%). Canagliflozin reduced the risk of major cardiovascular events overall (hazard ratio [HR], 0.80 [95% CI, 0.67-0.95]; P=0.01), with consistent reductions in both the primary (HR, 0.68 [95% CI, 0.49-0.94]) and secondary (HR, 0.85 [95% CI, 0.69-1.06]) prevention groups (P for interaction=0.25). Effects were also similar for the components of the composite including cardiovascular death (HR, 0.78 [95% CI, 0.61-1.00]), nonfatal myocardial infarction (HR, 0.81 [95% CI, 0.59-1.10]), and nonfatal stroke (HR, 0.80 [95% CI, 0.56-1.15]). The risk of the primary composite renal outcome and the composite of cardiovascular death or hospitalization for heart failure were also consistently reduced in both the primary and secondary prevention groups (P for interaction >0.5 for each outcome). Conclusions: Canagliflozin significantly reduced major cardiovascular events and kidney failure in patients with type 2 diabetes mellitus and chronic kidney disease, including in participants who did not have previous cardiovascular disease

Canagliflozin and renal outcomes in type 2 diabetes and nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to <90 ml per minute per 1.73 m2 of body-surface area and albuminuria (ratio of albumin [mg] to creatinine [g], >300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of <15 ml per minute per 1.73 m2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P<0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P<0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years

Apports récents de la pharmacologie des traitements immunosuppresseurs utilisés en transplantation d’organe

L’individualisation du traitement immunosuppresseur fait intervenir le suivi thérapeutique pharmacologique (STP) par la détermination du meilleur point unique ou la réalisation d’aires sous la courbe abrégées (abbreviated area under the curve-AUC), établies à partir des concentrations sanguines d’un médicament. Des travaux récents ont montré que la concentration de ciclosporine deux heures après la prise était le meilleur point unique. Cependant, une analyse critique laisse planer un doute sur l’intérêt clinique de cette approche. Les cibles de l’AUC de l’acide mycophénolique sont comprises entre 30 et 60 mg.h/l, mais les résultats cliniques discordants de deux études randomisées ne permettent pas de donner des conclusions claires sur son intérêt clinique. La pharmacogénétique est l’étude de l’influence de polymorphismes génétiques sur la réponse individuelle à un traitement. Il peut s’agir d’une réponse pharmacocinétique ou pharmacodynamique en terme d’efficacité ou de tolérance. Pour le tacrolimus, les posologies nécessaires pour obtenir les concentrations souhaitées sont corrélées à l’expression du cytochrome P450 3A5 (allèle *1). Une étude prospective de pharmacogénétique est en cours pour démontrer l’utilité de cette approche