Mechanisms of Heart Failure in HIV/AIDS: Nucleotides and NRTIs

HIV/艾滋病导致心力衰竭的机制：核苷酸和 NRTI

• 批准号: 8915899
• 负责人: WILLIAM LEWIS
• 金额: $ 63.77万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-08 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8915899
• 关键词: AIDS/HIV problem; Address; Adverse effects; Affect; Anti-Retroviral Agents; Antioxidants; Area; Attenuated; Biological; Biological Assay; Biological Factors; Cardiac; Cardiomyopathies; Cells; Chemicals; Congestive Heart Failure; Cytoplasm; Cytoplasmic Protein; DNA; DNA biosynthesis; Development; Dose; Equilibrium; Event; Exhibits; Functional disorder; Gene Transfer Techniques; Genetically Engineered Mouse; HIV; HIV-1; Health; Heart; Heart failure; In Vitro; Infection; Intake; Knockout Mice; Libraries; Manganese; Metabolism; Mitochondria; Mitochondrial DNA; Mitochondrial Proteins; Mus; Muscle Cells; Mutation; Myocardial; Myocardium; Nucleosides; Nucleotide Biosynthesis; Nucleotides; Organ; Oxidants; Oxidative Stress; Pathogenesis; Patients; Perfusion; Phenotype; Phosphorylation; Physiological; Porphyrins; Prevalence; Proteins; Pump; Purines; Pyrimidine; Resources; Resveratrol; Reverse Transcriptase Inhibitors; Ribonucleotide Reductase; Ribonucleotide Reductase Inhibitor; Role; SAM Domain; SOD2 gene; Sarcoplasm; Supplementation; Syndrome; Therapeutic; Therapeutic Agents; Toxic effect; Transgenic Mice; Transgenic Organisms; Work; base; catalase; catalyst; clinically relevant; heart cell; improved; in vivo; overexpression; prevent; purine; replicase; research study; tool; tripolyphosphate

DESCRIPTION (provided by applicant): This project defines how metabolism of nucleoside reverse transcriptase inhibitors (NRTIs) for HIV/AIDS causes defective mitochondrial (mt-) DNA replication, oxidative stress and cardiomyopathy (CM, a weakness of the heart muscle with poor pump function). Along with the benefit of increased survival in HIV/AIDS, combinations that include NRTIs carry devastating mitochondrial side effects, including CM. Mitochondrial (mt-) DNA depletion and mutation, and oxidative stress are key mechanisms in CM and CHF, and for development of NRTI toxic mitochondrial side effects based of defective mtDNA replication. The working hypothesis states: In myocardial cells, NRTIs interact with proteins that control the nucleotide biosynthesis and salvage for mtDNA replication. Ribonucleotide reductase (RNR) and SAM domain and HD domain-containing protein 1 (SAMHD1) alter abundance of dNTPs for mtDNA replication. RNR inhibition or increased activity of SAMHD1 depletes native dNTP pools, increases abundance of NRTI-TPs (or both). mtDNA depletion and mutation, oxidative stress, and CM are the phenotypic signature. Through clinically relevant experiments using genetically engineered mice, the project will dissect nucleotide pool imbalance in the sarcoplasm and mitochondria of CM myocytes in HIV/AIDS. Oxidative stress and CM are ameliorated by protecting cardiac mitochondria with manganese porphyrin catalytic antioxidants that localize to mitochondria. AIM 1: To define the role of RNR activity and inhibition in modulating cardiac cytoplasmic and mitochondrial nucleotide pools in the CM of HIV/AIDS. Transgenic mice (TGs) that overexpress RNR are in the lab presently. In heart cells, RNR overexpression (by transgenesis) or its inhibition (e.g., exogenous resveratrol) alters nucleotide pools for phosphorylation and incorporation into mtDNA. Nucleotide pools are defined with dNTP assays and correlated with cardiac physiological, pathological and pharmacological studies. AIM 2: To define the role of SAMHD1 activity (and absence) in modulating the myocardial cytoplasmic and mitochondrial nucleotide pools in the CM of HIV/AIDS. SAMHD1 null mice are powerful biological tools to increase the pool of dNTPs in heart cells. Conversely, cardiac targeted transgenic overexpression of SAMHD1 increases dN pools. NRTI treatment generates NRTI-TPs that inhibit pol ? and create CM. Cardiac nucleotide pools are determined with dNTP assays and correlative physiological, pathological and pharmacological parameters. AIM 3: To prevent and treat CM in HIV/AIDS with "replacement" by ameliorating mitochondrial oxidative stress. "Replacement of function" is accomplished using TGs from AIM 1 and 2 that are co- administered manganese porphyrin catalytic antioxidants (to provide SOD2- and catalase-like activity that bolsters antioxidant defense) and prevent or attenuate RNR- or SAMHD1-induced cardiac changes and attenuate CM pathophysiological changes.

描述（由申请人提供）：该项目定义了核苷逆转录酶抑制剂（NRTIS）的代谢如何导致有缺陷的线粒体（MT-）DNA复制，氧化应激，氧化应激和心肌病（CM，CM，CM，心脏较差的泵浦肌肉的弱点）。在艾滋病毒/艾滋病中生存增加的好处外，包括NRTI的组合携带了毁灭性的线粒体副作用，包括CM。线粒体（MT-）DNA耗竭和突变以及氧化应激是CM和CHF的关键机制，以及基于有缺陷的mtDNA复制的NRTI毒性线粒体副作用的发展。工作假设指出：在心肌细胞中，NRTIS与控制mtDNA复制的核苷酸生物合成和挽救的蛋白质相互作用。核糖核苷酸还原酶（RNR）和SAM结构域和含HD结构域的蛋白1（SAMHD1）改变了DNTP的丰度用于mtDNA复制。 RNR抑制或增加的SAMHD1活性耗尽天然DNTP池，增加了NRTI-TPS的丰度（或两者兼有）。 mtDNA耗竭和突变，氧化应激和CM是表型特征。通过使用基因工程小鼠进行临床相关的实验，该项目将剖析HIV/AIDS中CM肌细胞的核苷酸和线粒体中的核苷酸池失衡。通过用锰卟啉催化抗氧化剂保护心脏线粒体，可以改善氧化应激和CM，该氧化应力定位于线粒体。目标1：定义RNR活性和抑制在调节HIV/AIDS CM中心脏细胞质和线粒体核苷酸池中的作用。过表达RNR的转基因小鼠目前在实验室中。在心脏细胞中，RNR过表达（通过转基因）或其抑制作用（例如，外源白藜芦醇）改变了核苷酸池，以使磷酸化并掺入mtDNA中。核苷酸池用DNTP测定法定定义，并与心脏生理，病理和药理研究相关。目标2：定义SAMHD1活性（和缺失）在调节艾滋病毒/艾滋病CM中的心肌细胞质和线粒体核苷酸池中的作用。 SAMHD1 NULL小鼠是增加心脏细胞中DNTP池的强大生物学工具。相反，SAMHD1的心脏靶向转基因过表达增加了DN池。 NRTI处理会产生抑制POL的NRTI-TPS？并创建CM。通过DNTP测定以及相关生理，病理和药理参数确定心脏核苷酸池。目标3：通过改善线粒体氧化应激，以“替代”预防和治疗艾滋病毒/艾滋病中的CM。使用AIM 1和2的TGS来完成“功能的替代”，这些TG是由共同给药的锰卟啉催化抗氧化剂（提供促进抗氧化剂防御的SOD2-和过氧化氢酶样活性）并预防或减轻RNR-或SAMHD1诱导的心脏诱导的心脏变化和促成CM疗法的pathysioly pathysioly pathypaterylycy listherally。