Developmental Pharmacology of Antiretroviral Metabolism in Mucosal Tissues

粘膜组织中抗逆转录病毒代谢的发育药理学

• 批准号: 9244420
• 负责人: Namandje N Bumpus
• 金额: $ 51.72万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-10 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9244420
• 关键词: Address; Adherence; Adolescent; Adult; Anti-HIV Agents; Anti-Retroviral Agents; Bioinformatics; Biological Assay; Biopsy; Blood; Cells; Clinical Trials; Colorectal; Data; Development; Diphosphates; Dose; Drug Exposure; Drug Kinetics; Drug usage; Enzymes; Erythrocytes; Estradiol; Exhibits; Failure; Female; Foundations; Goals; HIV; HIV Infections; HIV antiretroviral; Hormonal; Hormonal Change; Hormones; Image; In Situ; Individual; Investigation; Knowledge; Liver; MM form creatine kinase; Mass Spectrum Analysis; Measures; Metabolic Biotransformation; Metabolism; Methodology; Mind; Mucous Membrane; Muscle; Nucleotides; Pathway interactions; Pattern; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Pharmacology; Phosphorylation; Phosphotransferases; Positioning Attribute; Production; Progesterone; Proteomics; Pyruvate Kinase; Regulation; Reporting; Reverse Transcriptase Inhibitors; Site; Spatial Distribution; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tenofovir; Testing; Testosterone; Time; Tissues; Vagina; Woman; Work; adenylate kinase; base; enzyme activity; experience; genetic variant; hormone regulation; improved; insight; interest; knock-down; lens; male; men; pre-exposure prophylaxis; predictive tools; prevent; protein activation; reproductive; reproductive development; reproductive hormone; response; sex; sexual dimorphism; tool; transcription factor

Tenofovir (TFV) and other nucleotide reverse transcriptase inhibitors must be activated via phosphorylation by intracellular nucleotide kinases in order to become pharmacologically active. While the kinases that activate TFV in mucosal tissues in adults have been identified, studies have not been performed to determine which enzymes phosphorylate TFV in adolescents. Further, whether hormonal changes during development might impact the expression and activity of the kinases that have thus far been demonstrated to activate TFV hasn't been explored. Gaining a mechanistic understanding of the expression patterns and regulation of these nucleotide kinases across developmental stages in mucosal tissues is of particular importance within the context of using these drugs for HIV pre-exposure prophylaxis since vaginal and colorectal tissues are putative sites of pharmacological activity in this setting. With this in mind, the goals of this proposal are to identify the nucleotide kinases that phosphorylate TFV in adolescent vaginal and colorectal tissue and to probe the effects of hormones that govern developmental maturation on TFV disposition. The aims are as follows: (1) determine which kinases activate TFV in adolescent tissue; the expression pattern of nucleotide kinases in adolescent vaginal and colorectal tissue will be established using proteomics-based approaches; knockdown of identified kinases will facilitate identification of those that exhibit activity toward TFV in tissue; (2) test whether TFV activation and TFV distribution in tissue are hormonally regulated; nucleotide kinase expression will be measured in response to developmental hormones and mechanisms of hormonal regulation will be probed; MALDI-mass spectrometry imaging will be employed in order to visualize the distribution of TFV as well as phosphorylated metabolites of TFV in adolescents versus adults. It is expected that the completion of the proposed studies will provide a mechanistic foundation for the rational selection and dosing of antiretrovirals for HIV pre-exposure prophylaxis in adolescents.

替诺福韦（TFV）和其他核苷酸逆转录酶抑制剂必须通过磷酸化被激活， 细胞内的核苷酸激酶，以成为具有生物活性的。而激活TFV的激酶 在成人的粘膜组织中已经鉴定出，尚未进行研究以确定哪些酶 磷酸化TFV。此外，发育过程中的激素变化是否会影响 迄今为止已证实激活TFV的激酶的表达和活性尚未被证实。 探讨了获得对这些核苷酸的表达模式和调控的机制性理解， 粘膜组织中跨越发育阶段的激酶在使用 这些药物用于HIV暴露前预防，因为阴道和结肠直肠组织是HIV感染的假定部位。 药理活性在这种情况下。考虑到这一点，本提案的目标是确定核苷酸 青少年阴道和结直肠组织中磷酸化TFV的激酶，并探讨激素的作用 支配TFV倾向的发育成熟的基因。目的如下：（1）确定哪些激酶 激活青春期组织中的TFV;青春期阴道和阴道组织中核苷酸激酶的表达模式， 将使用基于蛋白质组学的方法建立结直肠组织;敲除鉴定的激酶将 有助于鉴定组织中对TFV表现出活性的那些;（2）测试TFV激活和TFV是否 在组织中的分布是受神经调节的;核苷酸激酶表达将响应于 将探讨发育激素和激素调节机制; MALDI质谱法 将采用成像以可视化TFV以及磷酸化代谢物的分布。 青少年与成人的TFV。预计拟议研究完成后， HIV暴露前预防用抗逆转录病毒药物的合理选择和剂量的机制基础 在青少年中。