Solid organ transplantation and gut microbiota: a review of the potential immunomodulatory properties of short-chain fatty acids in graft maintenance.

Transplantation is the treatment of choice for several end-stage organ defects: it considerably improves patient survival and quality of life. However, post-transplant recipients may experience episodes of rejection that can favor or ultimately lead to graft loss. Graft maintenance requires a complex and life-long immunosuppressive treatment. Different immunosuppressive drugs (i.e., calcineurin inhibitors, glucocorticoids, biological immunosuppressive agents, mammalian target of rapamycin inhibitors, and antiproliferative or antimetabolic agents) are used in combination to mitigate the immune response against the allograft. Unfortunately, the use of these antirejection agents may lead to opportunistic infections, metabolic (e.g., post-transplant diabetes mellitus) or cardiovascular (e.g., arterial hypertension) disorders, cancer (e.g., non-Hodgkin lymphoma) and other adverse effects. Lately, immunosuppressive drugs have also been associated with gut microbiome alterations, known as dysbiosis, and were shown to affect gut microbiota-derived short-chain fatty acids (SCFA) production. SCFA play a key immunomodulatory role in physiological conditions, and their impairment in transplant patients could partly counterbalance the effect of immunosuppressive drugs leading to the activation of deleterious pathways and graft rejection. In this review, we will first present an overview of the mechanisms of graft rejection that are prevented by the immunosuppressive protocol. Next, we will explain the dynamic changes of the gut microbiota during transplantation, focusing on SCFA. Finally, we will describe the known functions of SCFA in regulating immune-inflammatory reactions and discuss the impact of SCFA impairment in immunosuppressive drug treated patients.

Exploring mitochondrial metabolism of wild-type and diabetic mice skin explants using the Seahorse technology.

BACKGROUND: Skin wound healing is a complex mechanism which requires a lot of energy, mainly provided by mitochondrial respiration. However, little is known about the mitochondrial bioenergetics of mice skin. We sought to develop a microplate-based assay to directly measure oxygen consumption in whole mice skin with the goal of identifying mitochondrial dysfunction in diabetic skin using an extracellular flux. MATERIALS AND METHODS: Different parameters were optimized to efficiently measure the oxygen consumption rate (OCR). First, the most pertinent skin side of wild-type mice was first determined. Then, concentrations of mitochondrial inhibitors were then optimized to get the best efficacy. Finally, punch sizes were modulated to get the best OCR profile. RESULTS: Dermis had the best metabolic activity side of the skin. Unlike the increased concentrations of carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) and rotenone/antimycin A, which showed no improvement of these drugs' effects, varying the skin punch size was successful. Finally, type II diabetic (T2D) skin produced less ATP through mitochondrial metabolism and had a greater non-mitochondrial oxygen consumption than wild-type or type I diabetic (T1D) skin. CONCLUSION: Here we designed, for the first time, a reliable protocol to measure mitochondria function in whole mouse skin. Our optimized protocol was valuable in assessing alterations associated with diabetes and could be applied to future studies of pathological human skin metabolism.

ClC-K Kidney Chloride Channels: From Structure to Pathology.

The molecular basis of chloride transport varies all along the nephron depending on the tubular segments especially in the apical entry of the cell. The major chloride exit pathway during reabsorption is provided by two kidney-specific ClC chloride channels ClC-Ka and ClC-Kb (encoded by CLCNKA and CLCNKB gene, respectively) corresponding to rodent ClC-K1 and ClC-K2 (encoded by Clcnk1 and Clcnk2). These channels function as dimers and their trafficking to the plasma membrane requires the ancillary protein Barttin (encoded by BSND gene). Genetic inactivating variants of the aforementioned genes lead to renal salt-losing nephropathies with or without deafness highlighting the crucial role of ClC-Ka, ClC-Kb, and Barttin in the renal and inner ear chloride handling. The purpose of this chapter is to summarize the latest knowledge on renal chloride structure peculiarity and to provide some insight on the functional expression on the segments of the nephrons and on the related pathological effects.

Can precision medicine be integrated into routine therapeutic decisions at the bedside of patients?

Therapeutic strategies are shifting from a "one-size-fits-all" population-based approach to a stratified approach targeting groups with similar characteristics, or even individuals, tailoring treatments to the unique characteristics of each patient. Since such strategies rely on increasingly complex knowledge and healthcare technologies, along with an understanding of the tools of precision medicine, the appropriate dissemination and use of these strategies involves a number of challenges for the medical community. Having evaluation methodologies that have been jointly designed with the institutional, industrial, academic stakeholders, and also patients, like streamlining the processes and externally validating performances, could enhance the relevance of the "evaluation" aspect of precision medicine. Creating a network of expert precision-medicine centers and ensuring that precision-medicine procedures are reimbursed by social security would guarantee fair and sustainable access. Finally, training healthcare professionals, creating interfaces between precision-medicine expert centers and primary care professionals as well as patients, and integrating individual patient data into medical records are all key drivers that will enable information from precision-medicine to be made available and guarantee the proper use of these approaches.

Pendrin: linking acid base to blood pressure.

Pendrin (SLC26A4) is an anion exchanger from the SLC26 transporter family which is mutated in human patients affected by Pendred syndrome, an autosomal recessive disease characterized by sensoneurinal deafness and hypothyroidism. Pendrin is also expressed in the kidney where it mediates the exchange of internal HCO3- for external Cl- at the apical surface of renal type B and non-A non-B-intercalated cells. Studies using pendrin knockout mice have first revealed that pendrin is essential for renal base excretion. However, subsequent studies have demonstrated that pendrin also controls chloride absorption by the distal nephron and that this mechanism is critical for renal NaCl balance. Furthermore, pendrin has been shown to control vascular volume and ultimately blood pressure. This review summarizes the current knowledge about how pendrin is linking renal acid-base regulation to blood pressure control.

GDF15 mediates renal cell plasticity in response to potassium depletion in mice.

OBJECTIVE: To understand the mechanisms involved in the response to a low-K+ diet (LK), we investigated the role of the growth factor GDF15 and the ion pump H,K-ATPase type 2 (HKA2) in this process. METHODS: Male mice of different genotypes (WT, GDF15-KO, and HKA2-KO) were fed an LK diet for different periods of time. We analyzed GDF15 levels, metabolic and physiological parameters, and the cellular composition of collecting ducts. RESULTS: Mice fed an LK diet showed a 2-4-fold increase in plasma and urine GDF15 levels. Compared to WT mice, GDF15-KO mice rapidly developed hypokalemia due to impaired renal adaptation. This is related to their 1/ inability to increase the number of type A intercalated cells (AIC) and 2/ absence of upregulation of H,K-ATPase type 2 (HKA2), the two processes responsible for K+ retention. Interestingly, we showed that the GDF15-mediated proliferative effect on AIC was dependent on the ErbB2 receptor and required the presence of HKA2. Finally, renal leakage of K+ induced a reduction in muscle mass in GDF15-KO mice fed LK diet. CONCLUSIONS: In this study, we showed that GDF15 and HKA2 are linked and play a central role in the response to K+ restriction by orchestrating the modification of the cellular composition of the collecting duct.

Experience and expectations of pharmacogenetic tests in France.

Although French genomic medicine is reaching a turning point in its history and the implementation of genome sequencing in routine is being implemented as part of the France Genomic Medicine 2025 Plan (FGMP), many questions about secondary data management remain to be addressed. In particular, the use of pharmacogenetic (PGx) information that can be extracted from genome data is a concern. We sought to analyze the opinion of French health professionals on their desire to have access to this information. For this purpose, we created a 22-item questionnaire on the experiences, attitudes, expectations, and knowledge of French physicians and pharmacists about PGx. We collected the responses in different groups and determined a knowledge score with the last 3 questions of the questionnaire. Then, we built a prediction model for this score and determined which factors may influence it. Half of the responders were physicians (158/311) and the other half were pharmacists (153/311), and the majority of them worked in a hospital (265/311). Almost two third (62.7%, 195/311) of the responders thought that pharmacogenetic data should be communicated with genomic results for the primary indication within the framework of FGMP, and 89.1% (277/311) of them that PGx tests could be an interesting tool to optimize patients' drug therapy in the future. Only 11.2% (35/311) of the responders reached the maximum knowledge score, while 25.4% (76/311) had already prescribed or recommended a PGx test. This study identified a need for training for French physicians and pharmacists in PGx, particularly given the interest of health professionals in it.

Metabolic interactions of benzodiazepines with oxycodone ex vivo and toxicity depending on usage patterns in an animal model.

BACKGROUND AND PURPOSE: Opioids and benzodiazepines are frequently combined in medical as well as in non-medical contexts. At high doses, such combinations often result in serious health complications attributed to pharmacodynamics interactions. Here, we investigate the contribution of the metabolic interactions between oxycodone, diazepam and diclazepam (a designer benzodiazepine) in abuse/overdose conditions through ex vivo, in vivo and in silico approaches. EXPERIMENTAL APPROACH: A preparation of pooled human liver microsomes was used to study oxycodone metabolism in the presence or absence of diazepam or diclazepam. In mice, diazepam or diclazepam was concomitantly administered with oxycodone to mimic acute intoxication. Diclazepam was introduced on Day 10 in mice continuously infused with oxycodone for 15 days to mimic chronic intoxication. In silico modelling was used to study the molecular interactions of the three drugs with CYP3A4 and 2D6. KEY RESULTS: In mice, in acute conditions, both diazepam and diclazepam inhibited the metabolism of oxycodone. In chronic conditions and at pharmacologically equivalent doses, diclazepam drastically enhanced the production of oxymorphone. In silico, the affinity of benzodiazepines was higher than oxycodone for CYP3A4, inhibiting oxycodone metabolism through CYP3A4. Oxycodone metabolism is likely to be diverted towards CYP2D6. CONCLUSION AND IMPLICATIONS: Acute doses of diazepam or diclazepam result in the accumulation of oxycodone, whereas chronic administration induces the accumulation of oxymorphone, the toxic metabolite. This suggests that overdoses of opioids in the presence of benzodiazepines are partly due to metabolic interactions, which in turn explain the patterns of toxicity dependent on usage. LINKED ARTICLES: This article is part of a themed issue on Advances in Opioid Pharmacology at the Time of the Opioid Epidemic. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v180.7/issuetoc.

Randomized comparative double-blind study assessing the difference between topically applied microbiome supporting skincare versus conventional skincare on the facial microbiome in correlation to biophysical skin parameters.

BACKGROUND: There are trillions of live bacteria, of around 1000 different species, living in human skin which are considered essential for the balance and barrier function of the skin. The gut microbiome has been a subject of extensive research and evidence shows that the gut flora is affected by preservatives and processed foods. In conventional skincare, preservatives are used, and this raises the question of how it affects the skin flora and its balance. METHODS: A randomized double-blind study on 14 healthy volunteers ages 23-45 years old were advised to use microbiome-supporting (MS) products on one cheek and benchmark (BM) products on the other cheek daily for 3 weeks. To investigate how the skin was affected, the skin microbiome was analysed using 16 S rRNA sequencing and biophysical parameters were assessed using an Antera 3D camera. Measurements were performed before and after the 3 weeks of using the products. RESULTS: The use of MS products for 3 weeks significantly increased the total number of reads mapped to unique bacterial species (p < 0.05) and the number of different unique species (p < 0.05). In addition, the use of MS products significantly reduced redness (p < 0.05) and improved skin texture (p < 0.01). The use of BM products showed no significant difference in any of the parameters except improved skin texture (p < 0.05). Additionally, the MS side showed a significantly improved diversity (p < 0.05) compared with the BM side. The four major phyla found were, similarly to previous findings by others, Actinobacteria, Firmicutes, Proteobacteria and Bacteroidetes. Some of the most prevalent species were Cutibacterium acnes, Staphylococcus epidermidis and Pseudonomas aeruginosa. CONCLUSION: The findings of this study showed significant improvements in the microbiome and biophysical parameters within 3 weeks of using MS skincare alone, while BM skincare only gave significantly improved skin roughness. Importantly, the MS side gave a significantly improved bacterial Shannon diversity (p < 0.05) compared with the BM side. Regarding the biophysical parameters, the MS skincare gave significant improvements in several parameters compared with baseline. However, they were not yet significant when compared to using BM skincare and therefore a larger study population will be needed. Importantly, this is the first study to investigate how preservatives affect the facial microbiome in vivo and has raised a need for further investigation. These results together with further studies can lead to innovations within the cosmetic industry that promote healthier skin.

CONTEXTE: Il existe des milliers de milliards de bactéries vivantes, d'environ 1 000 espèces différentes, vivant dans la peau humaine, qui sont considérées comme essentielles pour l'équilibre et la fonction barrière de la peau. Le microbiome intestinal a fait l'objet de recherches approfondies, et des preuves montrent que la flore intestinale est affectée par les conservateurs et les aliments transformés. Dans les soins de la peau traditionnels, des conservateurs sont utilisés, ce qui soulève la question de leur effet sur la flore cutanée et son équilibre. MÉTHODES: Dans une étude randomisée en double aveugle, il a été conseillé à 14 volontaires en bonne santé âgés de 23 à 45 ans d'utiliser quotidiennement des produits nourrissant le microbiome (NM) sur une joue et des produits de référence sur l'autre joue pendant 3 semaines. Pour étudier l'effet sur la peau, le microbiome cutané a été analysé à l'aide d'un séquençage ARNr 16S, et les paramètres biophysiques ont été évalués à l'aide d'une caméra 3D Antera. Les mesures ont été réalisées avant et après les 3 semaines d'utilisation des produits. RÉSULTATS: L'utilisation de produits NM pendant 3 semaines a significativement augmenté le nombre total de résultats cartographiés pour des espèces bactériennes uniques (p < 0,05) et le nombre d'espèces uniques différentes (p < 0,05). En outre, l'utilisation de produits NM a significativement réduit la rougeur (p < 0,05) et amélioré la texture de la peau (p < 0,01). L'utilisation de produits de référence n'a montré aucune différence significative dans aucun des paramètres, à l'exception de l'amélioration de la texture de la peau (p < 0,05). En outre, les produits NM ont montré une diversité significativement améliorée (p < 0,05) par rapport aux produits de référence. Les quatre principaux embranchements identifiés étaient, de manière similaire aux résultats précédents d'autres études, les actinobactéries, les bacillota, les protéobactéries et les bactéroïdètes. Certaines des espèces les plus prévalentes étaient Cutibacterium acnes, Staphylococcus epidermidis et Pseudomonas aeruginosa. CONCLUSION: Les résultats de cette étude ont montré des améliorations significatives du microbiome et des paramètres biophysiques dans les 3 semaines suivant l'utilisation de soins de la peau NM seuls, tandis que les soins de la peau de référence n'ont permis qu'une amélioration significative de la rugosité de la peau. Il est important de noter que les produits NM ont permis une amélioration significative de la diversité bactérienne de Shannon (p < 0,05) par rapport aux produits de référence. En ce qui concerne les paramètres biophysiques, les soins de la peau NM ont apporté des améliorations significatives de plusieurs paramètres par rapport à l'entrée dans l'étude. Cependant, ils n'étaient pas encore significatifs par rapport à l'utilisation de soins de la peau de référence et, par conséquent, une étude avec une population plus importante sera nécessaire. Il est important de noter qu'il s'agit de la première étude examinant l'effet des conservateurs sur le microbiome facial in vivo et qu'elle a soulevé la nécessité d'une étude plus approfondie. Ces résultats, ainsi que des études complémentaires, peuvent conduire à des innovations au sein de l'industrie cosmétique qui favorisent une peau en meilleure santé.

Co-limitation towards lower latitudes shapes global forest diversity gradients.

The latitudinal diversity gradient (LDG) is one of the most recognized global patterns of species richness exhibited across a wide range of taxa. Numerous hypotheses have been proposed in the past two centuries to explain LDG, but rigorous tests of the drivers of LDGs have been limited by a lack of high-quality global species richness data. Here we produce a high-resolution (0.025° × 0.025°) map of local tree species richness using a global forest inventory database with individual tree information and local biophysical characteristics from ~1.3 million sample plots. We then quantify drivers of local tree species richness patterns across latitudes. Generally, annual mean temperature was a dominant predictor of tree species richness, which is most consistent with the metabolic theory of biodiversity (MTB). However, MTB underestimated LDG in the tropics, where high species richness was also moderated by topographic, soil and anthropogenic factors operating at local scales. Given that local landscape variables operate synergistically with bioclimatic factors in shaping the global LDG pattern, we suggest that MTB be extended to account for co-limitation by subordinate drivers.

Detection of relevant pharmacogenetic information through exome sequencing in oncology.

Background: Germline sequencing of individual genomes can detect alleles responsible for adverse drug reactions (ADRs) in relation to chemotherapy, targeted agents, antiemetics or pain treatment. Materials & methods: To evaluate the interest of such pharmacogenetic information, the authors retrospectively analyzed genes known to have an impact on cancer therapy in a cohort of 445 solid cancers patients. Results: Six patients treated with 5-fluorouracil carrying one DPYD variant classified as 1A showed decreased drug mean clearance (p = 0.01). Regarding CYP2D6, all patients (n = 5) with predicted CYP2D6 poor or ultra-rapid metabolizer status experienced adverse drug reactions related to opioid therapy. Conclusion: Genomic germline sequencing performed for theragnostic issues in patients with a solid tumor, can provide relevant information about common pharmacogenetic alleles.

Is Intestinal Dysbiosis-Associated With Immunosuppressive Therapy a Key Factor in the Pathophysiology of Post-Transplant Diabetes Mellitus?

Post-transplant diabetes mellitus (PTDM) is one of the most common and deleterious comorbidities after solid organ transplantation (SOT). Its incidence varies depending on the organs transplanted and can affect up to 40% of patients. Current research indicates that PTDM shares several common features with type 2 diabetes mellitus (T2DM) in non-transplant populations. However, the pathophysiology of PTDM is still poorly characterized. Therefore, ways should be sought to improve its diagnosis and therapeutic management. A clear correlation has been made between PTDM and the use of immunosuppressants. Moreover, immunosuppressants are known to induce gut microbiota alterations, also called intestinal dysbiosis. Whereas the role of intestinal dysbiosis in the development of T2DM has been well documented, little is known about its impacts on PTDM. Functional alterations associated with intestinal dysbiosis, especially defects in pathways generating physiologically active bacterial metabolites (e.g., short-chain fatty acids, trimethylamine N-oxide, indole and kynurenine) are known to favour several metabolic disorders. This publication aims at discussing the potential role of intestinal dysbiosis and dysregulation of bacterial metabolites associated with immunosuppressive therapy in the occurrence of PTDM.

Exome sequencing allows detection of relevant pharmacogenetic variants in epileptic patients.

Beyond the identification of causal genetic variants in the diagnosis of Mendelian disorders, exome sequencing can detect numerous variants with potential relevance for clinical care. Clinical interventions can thus be conducted to improve future health outcomes for patients and their at-risk relatives, such as predicting late-onset genetic disorders accessible to prevention, treatment or identifying differential drug efficacy and safety. To evaluate the interest of such pharmacogenetic information, we designed an "in house" pipeline to determine the status of 122 PharmGKB (Pharmacogenomics Knowledgebase) variant-drug combinations in 31 genes. This pipeline was applied to a cohort of 90 epileptic patients who had previously an exome sequencing (ES) analysis, to determine the frequency of pharmacogenetic variants. We performed a retrospective analysis of drug plasma concentrations and treatment efficacy in patients bearing at least one relevant PharmGKB variant. For PharmGKB level 1A variants, CYP2C9 status for phenytoin prescription was the only relevant information. Nineteen patients were treated with phenytoin, among phenytoin-treated patients, none were poor metabolizers and four were intermediate metabolizers. While being treated with a standard protocol (10-23 mg/kg/30 min loading dose followed by 5 mg/kg/8 h maintenance dose), all identified intermediate metabolizers had toxic plasma concentrations (20 mg/L). In epileptic patients, pangenomic sequencing can provide information about common pharmacogenetic variants likely to be useful to guide therapeutic drug monitoring, and in the case of phenytoin, to prevent clinical toxicity caused by high plasma levels.

Alternative to animal experimentation in pharmacology teaching: Development and validation of an equivalent digital learning tool.

Regarding animal experiments in pharmacology teaching, ethical considerations led us to examine an alternative approach to the use of living animals. This study aimed to assess whether digital tools could replace live animal experiments in terms of motivation and knowledge acquisition. The study was carried out with students enrolled in the 5th year of the industry/research stream at the Faculty of Pharmacy of the University of Limoges. The participants were randomly assigned to groups of traditional or digital teaching methods, with the common theme of the class being the effect of a diuretic agent (furosemide) in rats. The scenario and learning objectives were identical for the two groups. Before the class and after randomization, the acceptance of the digital educational material was assessed with a scale, which predicts the acceptability of users according to individual dimensions and social representations, followed by the assessment of the motivation by a situational motivation scale (SIMS) for both groups. After the class, the students' motivation was assessed by a questionnaire based on Deci and Ryan's self-determination theory. In the end, the participants were evaluated for homogeneity, based on general knowledge of renal pharmacology, and for knowledge acquisition concerning specific knowledge related to this teaching session. This study revealed a good acceptance of the digital tool and a good motivation toward the digital method among all the students. It found the two teaching methods (digital and traditional) to be equivalent in terms of motivation and knowledge acquisition. In our study, digital pedagogical tools as an alternative to live animals did not affect students' motivation and knowledge acquisition.

Identification of Workers at Increased Risk of Infection During a COVID-19 Outbreak in a Meat Processing Plant, France, May 2020.

On 13 May 2020, a COVID-19 cluster was detected in a French processing plant. Infected workers were described. The associations between the SARS-CoV-2 infection and the socio-demographic and occupational characteristics were assessed in order to implement risk management measures targeting workers at increased risk of contamination. Workers were tested by RT-PCR from samples taken during screening campaigns. Workers who tested positive were isolated and their contacts were quarantined. Workers were described and associations with the SARS-CoV-2 infection were assessed through risk ratios using multivariable Poisson regression. Of the 1347 workers, 87.5% were tested: 140 cases were identified; 4 were hospitalised, including 2 admitted to intensive care. In the company, the cluster remained limited to deboning and cutting activities. The attack rate was 11.9% in the company, reaching 16.6% in the cutting department. Being an employee of a subcontractor significantly increased the risk of infection by 2.98 [1.81-4.99]. In the cutting department, an association with virus infection was found for a group of non-French speaking workers from the same Eastern European country (RR = 2.67 [1.76-4.05]). They shared accommodation or carpooled more frequently than the other cases. The outbreak investigation revealed a significantly increased risk of SARS-CoV-2 infection for workers of subcontractors and some foreign-born workers. There are many such populations in meat processing plants; the observed associations and the ways in which these workers are contaminated need to be confirmed by further work. Prevention campaigns should now target these workers. Environmental risk factors in the workplace setting remain to be clarified.

The role of spatial competitive interactions between trees in shaping forest patterns.

General patterns that are observed in forests irrespective of their ecological biome or species composition are indicators of general biological processes that drive forest dynamics. Modelling can be used to infer these processes by identifying the minimum set of rules that generate the observed patterns. The regular spatial pattern of the largest trees, the decreasing skewness of the tree diameter distribution in young developing stands, and the size bimodality that may result from the tail of dominated trees are three general patterns observed in natural forests. Using a simple individual-based space-dependent growth model based on asymmetric competition between trees and space-independent approximations of this model, the objectives were (1) to identify a single mechanism that may explain these three patterns, and (2) to clarify the role of space in the emergence of these patterns. The space-dependent model was able to qualitatively generate the three patterns. Through cascading spatial interactions, competition resulted in the establishment of a spatially structured competitive hierarchy among trees. A second-order approximation of the space-dependent model was derived by structuring the population of trees according to diameter and competitive status. This second-order approximation succeeded in predicting left-skewed diameter distributions but did not predict diameter bimodality. Asymmetric competition thus seems to be an important driver of tree growth and the cause rather than the consequence of bimodality. The second-order approximation of the space-dependent model may be useful to generalize the stationary diameter distributions developed in the demographic equilibrium theory.

Acceptability of Pharmacogenetic Testing among French Psychiatrists, a National Survey.

Psychiatric disorder management is based on the prescription of psychotropic drugs. Response to them remains often insufficient and varies from one patient to another. Pharmacogenetics explain part of this variability. Pharmacogenetic testing is likely to optimize the choice of treatment and thus improve patients' care, even if concerns and limitations persist. This practice of personalized medicine is not very widespread in France. We conducted a national survey to evaluate the acceptability of this tool by psychiatrists and psychiatry residents in France, and to identify factors associated with acceptability and previous use. The analysis included 397 observations. The mean acceptability score was 10.70, on a scale from 4 to 16. Overall acceptability score was considered as low for 3.0% of responders, intermediate for 80.1% and high for 16.9%. After regression, the remaining factors influencing acceptability independently of the others were prescription and training history and theoretical approach. The attitude of our population seems to be rather favorable, however, obvious deficiencies have emerged regarding perceived skills and received training. Concerns about the cost and delays of tests results also emerged. According to our survey, one of the keys to overcoming the barriers encountered in the integration of pharmacogenetics seems to be the improvement of training and the provision of information to practitioners.

Personalized Therapy for Mycophenolate: Consensus Report by the International Association of Therapeutic Drug Monitoring and Clinical Toxicology.

ABSTRACT: When mycophenolic acid (MPA) was originally marketed for immunosuppressive therapy, fixed doses were recommended by the manufacturer. Awareness of the potential for a more personalized dosing has led to development of methods to estimate MPA area under the curve based on the measurement of drug concentrations in only a few samples. This approach is feasible in the clinical routine and has proven successful in terms of correlation with outcome. However, the search for superior correlates has continued, and numerous studies in search of biomarkers that could better predict the perfect dosage for the individual patient have been published. As it was considered timely for an updated and comprehensive presentation of consensus on the status for personalized treatment with MPA, this report was prepared following an initiative from members of the International Association of Therapeutic Drug Monitoring and Clinical Toxicology (IATDMCT). Topics included are the criteria for analytics, methods to estimate exposure including pharmacometrics, the potential influence of pharmacogenetics, development of biomarkers, and the practical aspects of implementation of target concentration intervention. For selected topics with sufficient evidence, such as the application of limited sampling strategies for MPA area under the curve, graded recommendations on target ranges are presented. To provide a comprehensive review, this report also includes updates on the status of potential biomarkers including those which may be promising but with a low level of evidence. In view of the fact that there are very few new immunosuppressive drugs under development for the transplant field, it is likely that MPA will continue to be prescribed on a large scale in the upcoming years. Discontinuation of therapy due to adverse effects is relatively common, increasing the risk for late rejections, which may contribute to graft loss. Therefore, the continued search for innovative methods to better personalize MPA dosage is warranted.

MRP4 is responsible for the efflux transport of mycophenolic acid ß-d glucuronide (MPAG) from hepatocytes to blood.

Mycophenolic acid (MPA) has become a cornerstone of immunosuppressive therapy, in particular for transplant patients. In the gastrointestinal tract, the liver and the kidney, MPA is mainly metabolized into phenyl-ß-d glucuronide (MPAG). Knowledge about the interactions between MPA/MPAG and membrane transporters is still fragmented. The aim of the present study was to explore these interactions with the basolateral hepatic MRP4 transporter. The inhibition of the MRP4-driven transport by various drugs which can be concomitantly prescribed was also evaluated. In vitro experiments using vesicles overexpressing MRP4 showed an ATP-dependent transport of MPAG driven by MRP4 (Michaelis-Menten constant of 233.9 ± 32.8 µM). MPA was not effluxed by MRP4. MRP4-mediated transport of MPAG was inhibited (from -43% to -84%) by ibuprofen, cefazolin, cefotaxime and micafungin. An in silico approach based on molecular docking and molecular dynamics simulations rationalized the mode of binding of MPAG to MRP4. The presence of the glucuronide moiety in MPAG was highlighted as key, being prone to make electrostatic and H-bond interactions with specific residues of the MRP4 protein chamber. This explains why MPAG is a substrate of MRP4 whereas MPA is not.