Mitochondrial Nucleotide Carriers of NRTI Metabolites

NRTI 代谢物的线粒体核苷酸载体

基本信息

批准号：
7275281
负责人：
TOD GULICK
金额：
$ 36.91万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-09-30 至 2009-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7275281
关键词：
ATP phosphohydrolase Acquired Immunodeficiency Syndrome Address Adenine Adenine Nucleotides Adverse effects Affinity Amino Acids Antiviral Agents Biochemical Genetics Biological Assay Carboxylic Acids Cardiac Cardiomyopathies Carnitine Carrier Proteins Cataloging Catalogs Cell Line Characteristics Chronic Citrate Citrates Clinical Coenzyme A Complement DNA Maintenance DNA biosynthesis DNA chemical synthesis DNA-Directed DNA Polymerase Deoxyribonucleotides Developmental Therapeutics Program Diphosphates Disruption Electron Transport Electrons Equilibrium Eukaryota Eukaryotic Cell Exclusion Failure Family Family member Fatty Acids Fishes Flavin Mononucleotide Folate Generations Genes Genetic Glutamine Goals HIV Heart Hereditary Disease Human In Vitro Kinetics LGLA Lactic Acidosis Light Liposomes Maintenance Malates Mammalian Cell Measures Mediating Membrane Metabolism Methionine Mitochondria Mitochondrial DNA Mitochondrial Proteins Mus Muscle Fibers Myopathy Neuropathy Nicotinamide adenine dinucleotide Nucleoside Transporter Nucleosides Nucleotides Numbers Oligopeptides Ornithine Orphan Orthologous Gene Parents Pathway interactions Patients Peripheral Nervous System Diseases Pharmaceutical Preparations Phenotype Physiological Plants Play Polymerase Chain Reaction Post-Translational Protein Processing Principal Investigator Prodrugs Production Protein Biosynthesis Protein Overexpression Proteins Proteolysis Protons RNA Interference RNA-Directed DNA Polymerase Radio Rate Resistance Respiratory Failure Respiratory physiology Retroviridae Reverse Transcriptase Inhibitors Rodent Role Saccharomyces cerevisiae Structure Symptoms System Testing Therapeutic Index Time Tissues Toxic effect Transgenes Viral Viral Physiology Yeasts Zalcitabine Zidovudine analog chemical group cofactor cytochrome c fly human prostaglandin D2 receptor hydroxyl group improved in vivo indexing malate man member mitochondrial dysfunction mitochondrial genome mutant novel nucleoside analog oxidation programs proteoliposomes radiochemical reconstitution respiratory response small molecule sugar synthetic nucleic acid tripolyphosphate uptake zidovudine triphosphate

项目摘要

DESCRIPTION (provided by applicant): Nucleoside reverse transcriptase inhibitors (NRTI) are critical components of highly active anti-viral therapies. Phosphorylated derivatives (NRTI-P) mimic the cellular dNTP substrates for HIV RT, but produce chain termination, thereby limiting retroviral amplification. NRTIs produce reversible side effects that resemble those of patients with mitochondrial (mito) genetic disorders, including cardiomyopathy, myopathy, neuropathy and lactic acidosis, and these correlate with NRTI-induced loss of mito DNA in various tissues. Toxicity has been attributed to inhibition of DNA polymerase g, interfering with mito DNA maintenance. Alternative or additional mechanisms of mito damage may also be at play. Regardless of mechanism, NRTI toxicity is dependent on import into mito. We have isolated novel human genes and cDNAs that encode orphan members of the mito metabolite carrier family (OMC). Five carry a signature specific to nucleotide (nt) carriers, including a putative adenine nt transporter (ANT), two putative CoA transporters, and a non-specific exchanger of nucleotides (OMC27). We have shown that purified OMC27 and its yeast ortholog transport NRTI-P's AZT-TP and -DP, ddl-TP, and ddC-TP in proteoliposome radiochemical flux assays. Disruption of the orthologous yeast gene protects against AZT-induced mito dysfunction. This project examines the role of OMC27 in NRTI-mediated toxicity, with goals of understanding mechanisms by which NRTIs gain entry to the matrix, and identifying nucleoside analogs with known anti-viral activity that offer potential for improved therapeutic index by virture of mito exclusion and reduced mito toxicity. We will test the hypotheses that OMC27 (and yeast ortholog) are highly affinity transporters of NRTI-P, that this represents the primary pathway for mito uptake of NRTI-P; that OMC27 expression is critical to NRTI-induced mito failure; and that a useful therapeutic index for NRTIs may be evident from the ratio of NRTI anti-viral to MC-mediated uptake activities; using complementary in vitro, yeast and mammalian cell, and in vivo mouse studies.

描述（由申请人提供）：核苷逆转录酶抑制剂（NRTI）是高效抗病毒疗法的关键成分。磷酸化衍生物 (NRTI-P) 模拟 HIV RT 的细胞 dNTP 底物，但会产生链终止，从而限制逆转录病毒扩增。 NRTI 会产生可逆的副作用，类似于线粒体 (mito) 遗传性疾病患者的副作用，包括心肌病、肌病、神经病和乳酸性酸中毒，这些副作用与 NRTI 诱导的各种组织中线粒体 DNA 的丢失有关。毒性归因于 DNA 聚合酶 g 的抑制，干扰线粒体 DNA 的维持。有丝分裂损伤的替代或附加机制也可能在发挥作用。无论机制如何，NRTI 毒性都依赖于进入线粒体。我们分离出了编码线粒体代谢物载体家族 (OMC) 孤儿成员的新型人类基因和 cDNA。其中五个携带核苷酸 (nt) 载体特有的特征，包括一个假定的腺嘌呤 nt 转运蛋白 (ANT)、两个假定的 CoA 转运蛋白和一个非特异性核苷酸交换蛋白 (OMC27)。我们已经证明，纯化的 OMC27 及其酵母直系同源物在蛋白脂质体放射化学通量测定中转运 NRTI-P 的 AZT-TP 和 -DP、ddl-TP 和 ddC-TP。直系同源酵母基因的破坏可以防止 AZT 诱导的有丝分裂功能障碍。该项目研究了 OMC27 在 NRTI 介导的毒性中的作用，目的是了解 NRTI 进入基质的机制，并鉴定具有已知抗病毒活性的核苷类似物，这些核苷类似物通过线粒体排斥和线粒体排斥而提供改善治疗指数的潜力。减少有丝分裂毒性。我们将测试以下假设：OMC27（和酵母直系同源物）是 NRTI-P 的高亲和力转运蛋白，这代表了有丝分裂摄取 NRTI-P 的主要途径； OMC27 表达对于 NRTI 诱导的有丝分裂失败至关重要； NRTI 的有用治疗指数可以从 NRTI 抗病毒活性与 MC 介导的摄取活性的比率中看出；使用互补的体外、酵母和哺乳动物细胞以及小鼠体内研究。