Studies of toxicities of HIV RT inhibitors to human mitochondrial DNA polymerase

HIV RT抑制剂对人线粒体DNA聚合酶的毒性研究

基本信息

批准号：
8035443
负责人：
Yuhui Yin
金额：
$ 25万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-03-01 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8035443
关键词：
Acidosis Acquired Immunodeficiency Syndrome Adverse effects Adverse reactions Antiviral Agents Binding Biological Cardiomyopathies Catalytic Domain Cells Clinical Complex DNA DNA Repair DNA Structure DNA biosynthesis DNA polymerase gamma DNA-Directed DNA Polymerase Defect Deoxyribose Discrimination Drug Interactions Drug toxicity Encephalopathies Epidemic Equilibrium Event Excision Exhibits Exonuclease Gene Mutation HIV Highly Active Antiretroviral Therapy Holoenzymes Human Hydroxyl Radical Knowledge Lactose Lamivudine Mitochondria Mitochondrial DNA Mitochondrial Diseases Molecular Mutation Myopathy Nucleosides Nucleotides Organelles Patients Pharmaceutical Preparations Play Polymerase Proteins RNA-Directed DNA Polymerase Reagent Resolution Reverse Transcriptase Inhibitors Role Site Staging Structure Study Section Substrate Specificity Tissues Toxic effect Viral Virus Diseases Virus Replication Zalcitabine Zidovudine base design drug structure fighting human DNA inhibitor/antagonist pol Gene Products repaired success transmission process tripolyphosphate

项目摘要

DESCRIPTION (provided by applicant): Tremendous success fighting HIV infection with highly active antiretroviral therapy turns AIDS into a manageable clinical entity. However, nucleoside reverse transcriptase inhibitors (NRTIs), the indispensable component of the treatment, have displayed severe adverse reaction mostly manifested as mitochondrial toxicities. The toxicities have been implicated to NRTIs inhibitory effect on human mitochondrial DNA polymerase gamma (DNA Pol 3). We propose to elucidate structural basis of NRTIs mitochondrial toxicity by conducting structural studies of human Pol 3 captured at various stages of DNA replication. Specifically, we will determine crystal structures of Pol 3 during replication and structures of a stalled replicating Pol 3 by NRTIs zalcitabine and AZT. Comparison of these structures will provide molecular basis for NRTIs mitochondrial toxicity. We have recently obtained crystals of human Pol 3 holoenzyme containing the catalytic and accessory subunits that diffracted to 3.1 E resolution. In addition, we have obtained crystals of Pol 3 complexed with a primer-template DNA duplex, as well as a complex of Pol 3-DNA with zalcitabine. Upon completion of the proposed studies, we will provide an array of atomic-resolution snapshots of Pol 3 caught in action of DNA replication or inhibition. Comparison of inhibitor interactions with human Pol 3 with that of HIV reverse transcriptase will provide invaluable guidance in design high potency and low toxic antiviral reagents. PUBLIC HELATH RELEVANCE: Tremendous success fighting HIV infection with highly active antiretroviral therapy turns AIDS into a manageable clinical entity. However, nucleoside reverse transcriptase inhibitors (NRTIs), the indispensable component of the treatment, have displayed severe adverse reaction mostly manifested as mitochondrial toxicities. As AIDS epidemic continues and significantly increased patients survival duration, the side effects due to long-term usage of NRTIs become increasingly more common. Studies have implicated the NRTIs adverse reaction to inhibition of human mitochondrial DNA polymerase that replicates and repairs mitochondrial DNA. Active DNA replication is critical to the integrity of the organelle. Interference with human Pol 3 activity causes mutations and DNA depletion, resulting myopathy, cardiomyopathy, lactose acidosis and encephalopathy. We propose to conduct structural studies of NRTIs inhibitory mechanism of human mitochondrial DNA polymerase. The results of the study will not only increase our understanding of mitochondrial DNA replication, but also will be invaluable to aid design of high potency and low toxic anti- HIV reagents.

描述（由申请人提供）：与高度活跃的抗逆转录病毒疗法作斗争HIV感染的巨大成功，将艾滋病变成了可管理的临床实体。然而，核苷逆转录酶抑制剂（NRTIS）是治疗中必不可少的成分，已显示出严重的不良反应，主要表现为线粒体毒性。毒性与NRTIS抑制作用有关人线粒体DNA聚合酶γ（DNA POL 3）的影响。我们建议通过在DNA复制的各个阶段进行人类POL 3的结构研究来阐明NRTIS线粒体毒性的结构基础。具体而言，我们将在复制和nrtis zalcitabine和azt的复制后复制和复制POL 3的结构中确定POL 3的晶体结构。这些结构的比较将为NRTIS线粒体毒性提供分子基础。我们最近获得了人类POL 3全酶的晶体，其中含有衍射为3.1 E分辨率的催化和辅助亚基。此外，我们获得了与引物 - 板DNA双链体复合的POL 3的晶体，以及与Zalcitabine的Pol 3-DNA复合物。拟议的研究完成后，我们将提供一系列原子分辨率的快照，其中POL 3以DNA复制或抑制作用捕获。将抑制剂与人POL 3与HIV逆转录酶的相互作用进行比较将为设计高效力和低毒抗病毒药试剂提供宝贵的指导。公共Helath的相关性：与高度活跃的抗逆转录病毒疗法作斗争的HIV感染的巨大成功使AIDS变成了可管理的临床实体。然而，核苷逆转录酶抑制剂（NRTIS）是治疗中必不可少的成分，已显示出严重的不良反应，主要表现为线粒体毒性。随着艾滋病流行的继续并显着增加了患者的生存持续时间，由于长期使用NRTI引起的副作用变得越来越普遍。研究表明，NRTIS对抑制人线粒体DNA聚合酶的不良反应，该聚合酶复制和维修线粒体DNA。主动DNA复制对于细胞器的完整性至关重要。干扰人POL 3活性会导致突变和DNA耗竭，导致肌病，心肌病，乳糖酸中毒和脑病。我们建议对人线粒体DNA聚合酶的NRTIS抑制机制进行结构研究。该研究的结果不仅会增加我们对线粒体DNA复制的理解，而且对于有助于设计高效力和低毒性抗HIV试剂的设计也是无价的。