Characterization of the membrane transporter OATP1A2 activity towards different classes of drugs

Les transporteurs membranaires sont des éléments importants dans le devenir, l’efficacité, et la toxicité du médicament. Ils influencent la pharmacocinétique et la pharmacodynamie de ces derniers. Plusieurs interactions médicamenteuses observées cliniquement sont attribuables à la fois aux enzymes responsables du métabolisme des médicaments et aux transporteurs membranaires. Il est connu qu’une variabilité existe entre différents individus dans la réponse à un médicament et les polymorphismes génétiques retrouvés dans les gènes codant pour les transporteurs membranaires peuvent partiellement expliquer cette variabilité. OATP1A2 est un transporteur membranaire exprimé sur des organes importants, comme le cerveau et le rein. Plusieurs médicaments utilisés en clinique sont des substrats d’OATP1A2 et l’expression localisée de ce transporteur suggère un rôle important dans le devenir du médicament. Donc, mon projet de doctorat consistait à caractériser l’activité d’OATP1A2 en relation avec ses substrats et inhibiteurs, et de plus, à évaluer l’impact de différents variants génétiques d’OATP1A2 sur leur transport. Dans le premier article, la rosuvastatine a été utilisée comme substrat-type pour étudier le transport d’OATP1A2. Les expériences ont été menées en introduisant la rosuvastatine en compétition avec différent β-bloqueurs, une classe de médicaments rapportée dans la littérature comme substrats d’OATP1A2. Parmi les β-bloqueurs évalués, le carvédilol était l’inhibiteur le plus puissant. Dans la deuxième partie de l’étude, des médicaments ayant une structure similaire au carvédilol, tels que les antidépresseurs tricycliques, ont été évalués quant à leur potentiel d’inhibition sur OATP1A2. Une relation structure-activité a été définie à l’aide de ces données. Nous avons démontré que des composés tricycliques avec une courte chaîne aliphatique pouvaient inhiber OATP1A2. Dans le deuxième article, OATP1A2 a été étudié en considérant son expression et son rôle au sein de la barrière hémato-encéphalique (BHE). Des études précédentes ont démontré qu’OATP1A2 est exprimé sur la membrane luminale des cellules endothéliales formant la BHE. Nos données démontrent que les triptans, une classe de médicaments couramment utilisées pour traiter la crise migraineuse, sont des substrats d’OATP1A2 et que les composés tricycliques identifiés comme inhibiteurs d’OATP1A2 dans nos études précédentes peuvent inhiber le transport des triptans par OATP1A2. Ces résultats sont importants puisque: 1) il a été suggéré que les triptans peuvent agir au niveau du système nerveux central en se liant aux récepteurs trouvés sur les neurones centraux; 2) comme les triptans sont des molécules hydrophiles, un mécanisme de transport facilité est nécessaire pour qu’ils pénètrent la BHE et OATP1A2 pourrait être l’élément clé; 3) l’inhibition d’OATP1A2 par les composés tricycliques pourrait limiter l’accès des triptans à leur site d’action. Le troisième article caractérise l’activité associée à deux variants génétiques d’OATP1A2 (OATP1A2*2 et *3). Leur capacité à transporter les triptans et leur potentiel d’inhibition par les médicaments tricycliques ont été évalués. Des résultats supplémentaires caractérisant OATP1A2, mais sans liens directs avec les trois articles, seront présentés en annexe. Dans l’ensemble, les résultats présentés dans cette thèse servent à caractériser le transporteur membranaire OATP1A2 en relation avec ses substrats et inhibiteurs, et en fonction de ses variants génétiques.Drug transporters are important determinants in drug disposition, efficacy, and toxicity. They influence the pharmacokinetics and pharmacodynamics of drugs. Several clinically-observed drug-drug interactions are mediated through drug metabolizing enzymes and drug transporters. It is well known that there is an interindividual variability in the response to medications and polymorphisms found in genes encoding for drug transporters partially account for it. OATP1A2 is a membrane drug transporter expressed on important organs, such as the brain and the kidney. A wide spectrum of drugs used in the clinic are substrates of OATP1A2. Its localisation suggests an essential role in drug disposition. Thus, my PhD project consisted of characterizing the activity of OATP1A2 in regards to its substrates, inhibitors, and different protein variants due to genetic polymorphisms. In the first article, rosuvastatin was used as the probe substrate to study OATP1A2 transport activity. Experiments were conducted by putting rosuvastatin in competition with different β-blockers, a class of drugs known in the literature to be transported by OATP1A2. One of the drugs evaluated, carvedilol, inhibited OATP1A2 with much more potency than the others. In the second part of the study, drugs with a structure similar to carvedilol, such as tricyclic antidepressants, were tested for their potential to inhibit OATP1A2. A structure-activity relationship was defined using the data. It was demonstrated that drugs composed of a tricyclic ring with a short aliphatic amine chain were potent OATP1A2 inhibitors. In the second article presented, OATP1A2 was studied in the context of its localization at the blood-brain barrier (BBB). OATP1A2 expression at the luminal membrane of the endothelial cells making up the BBB was demonstrated in the literature. Our article showed that triptans, a class of commonly used anti-migraine drugs, were OATP1A2 substrates. The tricyclic drugs previously evaluated were shown to potently inhibit triptan transport through OATP1A2. These findings are important for three reasons: 1) it has been postulated that triptans may act at the central nervous system by binding to receptors found on central neurons; 2) as triptans are hydrophilic molecules, a facilitated transport mechanism is required for them to penetrate the BBB and OATP1A2 may be the key player; and 3) the inhibition of OATP1A2 by the tricyclic drugs may limit the entrance of triptans to their site of action. The third article characterized the transport activity of two OATP1A2 protein variants (OATP1A2*2 and *3). Their capacities to transport triptans and their potential of being inhibited by tricyclic drugs were evaluated. Additional data characterizing OATP1A2 but considered out of the scope of the three articles will be presented in appendices. In overall, the central theme of this thesis looks into the characterization of the OATP1A2 membrane drug transporter in regards to its substrates, inhibitors, and proteins variants

Combination of product quality management and exports: The case of Colombian fruits companies

Quality management provides to companies a framework to improve quality in overall systems, reduction of costs, reallocation of resources efficiently, correct planning of strategies, prevent or correct errors in the right time and increase the performance of companies. In this text, we discuss the different theories in this field, their obligatory or non-obligatory compliance, the importance of quality management for exporting companies and a case study of a Colombian firm that its main objective is to manage quality. In conclusion, we find out that there is different types of quality management systems such as Juran’s trilogy, Deming 14 points, Six sigma, HACCP, and so on; also that companies have to manage suppliers and that quality has a positive influence on exports volume; in the case of Colombian small and medium enterprises, it can be mentioned that the majority has implemented tools regarding quality management but is not enough.Montpellier Business SchoolUniversidad del Rosari

Robust portfolio management with multiple financial analysts

Portfolio selection theory, developed by Markowitz (1952), is one of the best known and widely applied methods for allocating funds among possible investment choices, where investment decision making is a trade-off between the expected return and risk of the portfolio. Many portfolio selection models have been developed on the basis of Markowitz’s theory. Most of them assume that complete investment information is available and that it can be accurately extracted from the historical data. However, this complete information never exists in reality. There are many kinds of ambiguity and vagueness which cannot be dealt with in the historical data but still need to be considered in portfolio selection. For example, to address the issue of uncertainty caused by estimation errors, the robust counterpart approach of Ben-Tal and Nemirovski (1998) has been employed frequently in recent years. Robustification, however, often leads to a more conservative solution. As a consequence, one of the most common critiques against the robust counterpart approach is the excessively pessimistic character of the robust asset allocation. This thesis attempts to develop new approaches to improve on the respective performances of the robust counterpart approach by incorporating additional investment information sources, so that the optimal portfolio can be more reliable and, at the same time, achieve a greater return. [Continues.

Application of Genome Editing Technology to MicroRNA Research in Mammalians

Targeted nucleases have recently emerged as a powerful genome editing tool. The ability of introducing targeted, desired changes into mammalian genome makes them an invaluable tool to unravel functions of miRNAs in biology and disease. In combination with homologous donor vector, targeted nucleases can achieve high efficiency and precision, enabling bi-allelic ablation of miRNA in cultured somatic cells. Here we review the structure and function of miRNA as well as the unique implementation of genome editing technology in modifying miRNA sequences in mammalians. This chapter discusses the four mainstay genome editing technologies: meganuclease, zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN) and clustered regularly interspaced short palindromic repeat-associated nuclease Cas9 (CRISPR-Cas9), focusing on TALEN

Amyloid positron emission tomography candidates may focus more on benefits than risks of results disclosure

IntroductionGiven mounting calls to disclose biomarker test results to research participants, we explored factors underlying decisions by patients with mild cognitive impairment to receive amyloid imaging results.MethodsProspective, qualitative interviews were conducted with 59 participants (30 = mild cognitive impairment patients, 29 = care partners) from the scan arm of a randomized controlled trial on the effects of amyloid PET results disclosure in an Alzheimer Disease Research Center setting.ResultsSixty‐three percent of the participants were female, with an average age of 72.9 years, and most had greater than a high school level of education (80%). Primary motivations included: (1) better understanding one’s mild cognitive impairment etiology and prognosis to plan ahead, and (2) learning one’s brain amyloid status for knowledge’s sake, regardless of whether the information is actionable. Most participants demonstrated an adequate understanding of the scan’s limitations, yet instances of characterizing amyloid PET as a definitive test for Alzheimer’s disease occurred. Mention of potential drawbacks, such as negative psychological outcomes, was minimal, even among care partners.DiscussionFindings demonstrate a risk of disproportionate focus on possible benefits of testing among amyloid scan candidates and suggest a need to clearly emphasize the limitations of amyloid PET when counseling cognitively impaired patients and their families before testing. Future research should examine whether minimizing drawbacks at the pre‐imaging stage has adverse consequences on results disclosure.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/152635/1/dad2jdadm201805003.pd

Spin transport and accumulation in the persistent photoconductor Al0.3_{0.3}Ga0.7_{0.7}As

Electrical spin transport and accumulation have been measured in highly Si doped Al0.3Ga0.7As utilizing a lateral spin transport device. Persistent photoconductivity allows for the tuning of the effective carrier density of the channel material in situ via photodoping. Hanle effect measurements are completed at various carrier densities and the measurements yield spin lifetimes on the order of nanoseconds, an order of magnitude smaller than in bulk GaAs. These measurements illustrate that this methodology can be used to obtain a detailed description of how spin lifetimes depend on carrier density in semiconductors across the metal-insulator transition

SkewIT, Bracken, and Kraken: Methods for Analyzing a Complex, But Invisible World

As the DNA of the invisible world provides insight into the countless microscopic organisms living amongst us, the integrity of these genomes and the methods by which we analyze them become increasingly important. In the following, I introduce methods for both evaluating genomic integrity and analyzing microbial communities. For the analysis of bacterial genomes, I developed SkewIT (Skew Index Test) based on GC Skew, a bacterial genome phenomenon wherein the two replication strands of the same chromosome contain different proportions of guanine and cytosine nucleotides. SkewIT calculates a single metric representing the degree of GC skew for a single genome. Applied across 15,000+ complete bacterial genomes, SkewIT quickly detects assembly patterns and highlights potential bacterial mis-assemblies. Although eukaryotic microorganisms are abundant worldwide and as human pathogens, eukaryotic pathogen genomes are underrepresented in genomic databases and contain significant contamination. I therefore developed a bioinformatics system for eliminating contamination, generating a “clean” eukaryotic pathogen database (EuPathDB-Clean) of nearly 400 genomes. With the final database, I identify eukaryotic pathogens in human samples, demonstrating the increased sensitivity and reduction in false positives of the final database as compared to the originally contaminated genomes. As metagenomics captures the genomic data of all microbial organisms in any environment, I developed Bracken (Bayesian Reestimation of Abundance after Classification with KrakEN) for a quick and accurate characterization of the full microbial environment. Bracken uses the taxonomic assignments made by Kraken, a very fast read-level classifier, along with information about the genomes themselves to estimate abundance at the species level, the genus level, or above. I demonstrate that Bracken produces accurate abundance estimates even when a sample contains multiple near-identical species for both shotgun metagenomics projects and for 16S ribosomal RNA (rRNA) bacterial projects. SkewIT, Bracken, and EuPathDB-Clean are all publicly available for use in future metagenomics projects