Pharmacogenetics and Tramadol-Related Fatalities

Tramadol (TR) is a widely prescribed pain killer because of its relatively safe profile among opioids. Nevertheless, intoxication can occur and overdose can lead to fatal outcomes. Surprisingly, in some fatalities for which death is attributable to TR alone, postmortem blood concentration levels overlap with the therapeutic concentration range. These fatal cases might be explained by pharmacokinetic and pharmacodynamic properties of TR that are known to be both enantioselective and influenced by genes. Indeed pharmacogenetics (PG) is of great importance in this issue as it has the ability to elucidate the genetic variation contributing to drug absorption, distribution, metabolism, excretion, and response so that adverse drug reactions, toxicity, and even death can be avoided. The aim of this chapter is to present this issue

Identification of a Variable Number of Tandem Repeats Polymorphism and Characterization of LEF-1 Response Elements in the Promoter of the IDO1 Gene

Indoleamine 2,3-dioxygenase (IDO) catalyzes the first and rate-limiting step of the kynurenine pathway that is an important component of immunomodulatory and neuromodulatory processes. The IDO1 gene is highly inducible by IFN-γ and TNF-α through interaction with cis-acting regulatory elements of the promoter region. Accordingly, functional polymorphisms in the IDO1 promoter could partly explain the interindividual variability in IDO expression that has been previously documented.A PCR-sequencing strategy, applied to DNA samples from healthy Caucasians, allowed us to identify a VNTR polymorphism in the IDO1 promoter, which correlates significantly with serum tryptophan concentration, controlled partially by IDO activity, in female subjects, but not in males. Although this VNTR does not appear to affect basal or cytokine-induced promoter activity in gene reporter assays, it contains novel cis-acting elements. Three putative LEF-1 binding sites, one being located within the VNTR repeat motif, were predicted in silico and confirmed by chromatin immunoprecipitation. Overexpression of LEF-1 in luciferase assays confirmed an interaction between LEF-1 and the predicted transcription factor binding sites, and modification of the LEF-1 core sequence within the VNTR repeat motif, by site-directed mutagenesis, resulted in an increase in promoter activity.The identification of a VNTR in the IDO1 promoter revealed a cis-acting element interacting with the most downstream factor of the Wnt signaling pathway, suggesting novel mechanisms of regulation of IDO1 expression. These data offer new insights, and suggest further studies, into the role of IDO in various pathological conditions, particularly in cancer where IDO and the Wnt pathway are strongly dysregulated

Immune monitoring in melanoma and urothelial cancer patients treated with anti-PD-1 immunotherapy and SBRT discloses tumor specific immune signatures

Background: Blockade of the PD-1/PD-L1 pathway has revolutionized the oncology field in the last decade. However, the proportion of patients experiencing a durable response is still limited. In the current study, we performed an extensive immune monitoring in patients with stage III/IV melanoma and stage IV UC who received anti-PD-1 immunotherapy with SBRT. (2) Methods: In total 145 blood samples from 38 patients, collected at fixed time points before and during treatment, were phenotyped via high-parameter flow cytometry, luminex assay and UPLC-MS/MS. (3) Results: Baseline systemic immunity in melanoma and UC patients was different with a more prominent myeloid compartment and a higher neutrophil to lymphocyte ratio in UC. Proliferation (Ki67+) of CD8+ T-cells and of the PD-1+/PD-L1+ CD8+ subset at baseline correlated with progression free survival in melanoma. In contrast a higher frequency of PD-1/PD-L1 expressing non-proliferating (Ki67−) CD8+ and CD4+ T-cells before treatment was associated with worse outcome in melanoma. In UC, the expansion of Ki67+ CD8+ T-cells and of the PD-L1+ subset relative to tumor burden correlated with clinical outcome. (4) Conclusion: This study reveals a clearly different immune landscape in melanoma and UC at baseline, which may impact immunotherapy response. Signatures of proliferation in the CD8+ T-cell compartment prior to and early after anti-PD-1 initiation were positively correlated with clinical outcome in both cohorts. PD-1/PD-L1 expression on circulating immune cell subsets seems of clinical relevance in the melanoma cohort

Caractérisation de nouveaux variants alléliques de la Thiopurine S-Méthyltransferase (TPMT)

LILLE2-BU Santé-Recherche (593502101) / SudocSudocFranceF

Pharmacogénétique des traitements antiasthmatiques (analyse du polymorphisme génétique du récepteur des cystéinyl-leucotriènes de type 1 (CysLT1))

LILLE2-BU Santé-Recherche (593502101) / SudocSudocFranceF

aImplication du gène de l'indoleamine 2,3-dioxygénase (IDO1) dans les troubles comportementaux induits par l'interféron-a

L indoleamine 2,3-dioxygénase (IDO) catalyse la première étape limitante de la voie dite des kynurénines , qui est en compétition directe avec la voie de la sérotonine pour l utilisation dutryptophane. IDO est décrit comme étant un acteur important des phénomènes de neuromodulation et d immunorégulation. Des hypothèses avancées suggèrent que des variations d expression d IDO1 puissent s associer à un risque d apparition de maladies neuropsychiatriques (dont le syndrome dépressif), via une diminution de sérotonine d une part, et l augmentation de dérivés kynuréniques potentiellement neurotoxiques d autre part. L activité d IDO est fortement induite par l IFNg et l immunothérapie par IFNa entraîne le développement d effets indésirables d ordre neuropsychiatrique chez environ 30 % des patients traités. Nous émettons ainsi l hypothèse que des variations d expression et/ou d activité de l enzyme IDO, d origine génétique, puissent être impliquées dans la survenue de ces effets indésirables. Un polymorphisme de répétition du type VNTR ayant été identifié dans le promoteur d IDO1, un génotypage et un phénotypage d IDO ont étéréalisés chez 26 patients traités par l IFNa dans le cadre d une hépatite C, et ayant développé ou non des troubles neurocomportementaux pendant leur traitement. Cette approche in vivo a été complétée par une analyse fonctionnelle in vitro du VNTR, et par l étude de l effet de l IFNa sur l expression d IDO1 dans une lignée cellulaire humaine.LILLE2-BU Santé-Recherche (593502101) / SudocSudocFranceF

Development and validation of a GC-MS/MS method for the determination of ethylglucuronide in human urine and serum

Objectives: Ethyl-β-D-6-glucuronide (EtG) is a minor phase-II metabolite of ethanol. The aim of this work was to develop and validate a gas chromatography negative chemical ionization tandem mass spectrometry (GC-NCI-MS/MS) method to measure EtG levels in human urine and serum with both high sensitivity and specificity. Methods: EtG was extracted and purified from 1 mL urine or 0.5 mL serum by solid-phase extraction (SPE) using Mixed-mode Anion-eXchange (Oasis®MAX) extraction cartridges, followed by derivatization with pentafluoropropionic anhydride (PFPA). The analysis was performed in the multiple reaction monitoring (MRM) mode using the transitions m/z 496→163 (for EtG quantification), m/z 347→163 and m/z 496→119 (for identification), and m/z 501→163 for the internal standard EtG-D5. The validation procedure was performed according to the guidelines of the French Society of Analytical Toxicology (SFTA) and the French Committee of Accreditation (COFRAC; LAB GTA 04). Results: Calibration curves were linear in the concentration range of 10 to 10 000 ng/mL and 5 to 1 000 ng/mL in urine and serum, respectively, with a coefficient of correlation (r) above 0.996. The LOD and LOQ values were 5 and 10 ng/mL, respectively, for both matrices. The intra- and inter-day precision (relative standard deviation RSD%) and relative bias were less than 20%. Conclusion: To our knowledge, this is the first report of the application of a GC-MS/MS method for EtG measurement in urine and serum. The LOQ achieved appears to be better than those reported in the literature using other validated analytical techniques. This method could be used routinely for EtG measurement in various clinical and forensic contexts

Comments on "Evaluation and review of ways to differentiate sources of ethanol in post-mortem blood"

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Substance Use among Adolescents: A Retrospective Study (2017–2018) in the Toxicology Unit, University Hospital of Lille, France

Research on adolescent substance use is of utmost importance. Using local toxicological data, both prevalence and pattern of substance use (SU) and substance-related death (SRD) can be assessed to design effective prevention programs. A retrospective study of toxicology investigations of all adolescents referred to the medico-legal section of the Toxicology Unit of the University Hospital of Lille, France, for a 2-year period from 2017 to 2018. In the total sample of 1961 cases, adolescents accounted for 3.3% of the cases (n = 65). Among the adolescents, 16.9% were aged 10–14 years and 83.1% were aged >14–19 years. About 69.2% were males. Less than 70% of all presented adolescents used substances. More than two-thirds (74%) of positive detections were male. Illicit substances (43%) were the most detected substance followed by alcohol (20%) and prescription substances (20%). Tetrahydrocannabinol (THC) was extremely common as it was found in 29% of all adolescents. Cocaine and amphetamines were detected in 13.8% of total tested adolescents. Polysubstance use was common between alcohol and THC and among males. About one-third of deaths were due to substance use. About 54% of SRD was associated with polysubstance detection. It is recommended that illicit substances, ethanol, and prescription substances are targeted for testing among adolescents in order to provide appropriate prevention

Four-year follow-up of surface contamination by antineoplastic drugs in a compounding unit.

International audienceObjectives: This study aimed to monitor the contamination by antineoplastic drugs on work surfaces in a compounding unit 4 years after its implementation.Methods: This descriptive study was done in a unit performing on average 45 000 preparations per year. Surface sampling points (N=23) were monitored monthly in the frame of routine activity from the opening of an anticancer drug compounding unit. Contamination with nine antineoplastic drugs (cyclophosphamide, ifosfamide, dacarbazine, 5-fluorouracil, methotrexate, gemcitabine, cytarabine, irinotecan and doxorubicin) was assessed on wipes with a local liquid chromatography coupled with a tandem mass spectrometer analysis. The contamination rate (CR, %) was prospectively monitored every month during the entire study period. The occurrence of critical incidents was also registered. The effect of each safety measure implemented during this period was also analysed.Results: Based on the 1104 samples collected between March 2016 and March 2020, the CR was 18.5%. If three different critical incidents among a vial breakage that occurred were individually considered, this CR was slightly lower than that in the literature. Eight months after opening and taking different corrective actions, the overall CR dropped from 42.39% to 11.52% (p<0.001). Contamination was limited to the area that includes the compounding room and, more precisely, the welder and the QC-Prep+ sampling points.Conclusions: From the beginning of the study and from month to month, surface contamination was limited to the nearest sampling points to the compounding unit. This 4-year monitoring study allowed us to determine the intravenous conventional antineoplastic drugs and sampling points to be focused on.Keywords: Environmental Exposure; Health Personnel; Occupational Health; Public health