Mitochondrial DNA content in blood extracellular vesicles as a biomarker of neuronal mitochondrial DNA damage induced by cigarette smoking in virally suppressed, HIV-positive African Americans

血液细胞外囊泡中的线粒体 DNA 含量作为病毒抑制的 HIV 阳性非裔美国人吸烟引起的神经元线粒体 DNA 损伤的生物标志物

• 批准号: 10380583
• 负责人: Waldemar Popik
• 金额: $ 18.19万
• 依托单位: MEHARRY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10380583
• 关键词: Address; African American population; Biological Markers; Blood; Blood Cells; Blood Tests; Brain; Cells; Cerebrospinal Fluid; Cigarette Smoker; DNA; DNA Damage; Detection; Development; Diagnosis; Early Intervention; Functional disorder; General Population; Goals; HIV; HIV Infections; HIV Seronegativity; HIV Seropositivity; HIV diagnosis; HIV-associated neurocognitive disorder; High Prevalence; Impairment; Incidence; Individual; Intervention; Lead; Life Expectancy; Measures; Medical; Methods; Mitochondria; Mitochondrial DNA; Monitor; Nerve Degeneration; Neurocognitive; Neuronal Dysfunction; Neurons; Opportunistic Infections; Oxidative Phosphorylation; Oxidative Stress; Pathology; Persons; Ploidies; Population; Proteins; Public Health; RNA; RNA analysis; Research; Research Personnel; Sampling; Smoker; Smoking; Source; Spinal Tap; Surrogate Markers; System; Testing; Therapeutic Intervention; Time; Uncertainty; Viral; antiretroviral therapy; cigarette smoking; clinical application; cost; early detection biomarkers; extracellular vesicles; improved; innovation; mitochondrial dysfunction; nervous system disorder; non-smoker; novel; peripheral blood; real time monitoring; smoking prevalence; therapeutic development; wasting

Project Summary/Abstract Antiretroviral therapy (ART) drastically reduces the incidence of opportunistic infections and improves life expectancy among HIV-infected individuals. HIV-associated neurocognitive disorders (HAND) related to the use of ART remain highly prevalent (15–50%). Meanwhile, the diagnosis of HAND is often challenging. Moreover, there are no validated surrogate markers and definitive adjunctive treatment for HAND. Since the prevalence of smoking is three- to four-fold higher among HIV-infected individuals than in the general population, and that HIV-infected African Americans (AAs) represent 42% of individuals living with HIV in the U.S., there is an urgent need to address the effect of smoking on neuronal dysfunction in AAs living with HIV. Mitochondrial dysfunction is a hallmark of various neurological disorders. Mitochondrial damage and mitochondrial DNA (mtDNA) content reduction often occur prior to neuronal degeneration. Our long-term goal is to develop a clinically applicable, non-invasive test to monitor the effects of cigarette smoking on neuronal mtDNA content in HIV-positive individuals. This test will allow clinicians to monitor changes in neuronal mtDNA content in HIV-infected cigarette smokers on ART in order to facilitate early medical intervention. Our overall objective is to determine the effect of cigarette smoking on mtDNA content in neuron-derived extracellular vesicles (NEVs) isolated from the peripheral blood of AAs, according to smoking and HIV status. Our central hypothesis is that cigarette smoking exacerbates neuronal mtDNA damage in virally suppressed HIV-positive AAs, leading to increased release of mtDNA in peripheral blood NEVs. Our rationale is that mtDNA content in NEVs is a novel and non-invasive biomarker for early detection of mtDNA damage in neurons. Our specific aims are: 1) To quantify mtDNA content in NEVs isolated from the peripheral blood of AA non-smokers, including virally suppressed HIV-positive and HIV-negative subjects; and 2) To compare the mtDNA content in NEVs isolated from the peripheral blood of virally suppressed HIV-positive AA smokers with the mtDNA content in NEVs isolated from virally suppressed HIV-positive non-smokers, as well as HIV-negative smokers and non-smokers. In Aim #1, we will measure mtDNA content in NEVs isolated from the peripheral blood of HIV-negative and HIV-positive non-smokers by real-time quantitative PCR (qPCR) using intravesicular DNA extracted from NEVs. In Aim #2, we will analyze quantitative changes in mtDNA content in NEVs that may reflect mtDNA damage associated with cigarette smoking and ART. Our proposal is innovative because it proposes using mtDNA content in peripheral blood NEVs as a novel surrogate biomarker for monitoring neuronal mtDNA damage. Our findings from this proposed study will be significant they will establish mtDNA content in NEVs as a novel, non-invasive biomarker for real-time monitoring of neuronal dysfunction associated with mtDNA damage in HIV-positive individuals on ART and cigarette smokers.

项目总结/摘要 抗逆转录病毒疗法（ART）大大降低了机会性感染的发生率，改善了生活质量。 艾滋病毒感染者的预期。HIV相关的神经认知障碍（HAND） 抗逆转录病毒疗法的使用仍然非常普遍（15 - 50%）。同时，HAND的诊断往往具有挑战性。 此外，对于HAND，没有经过验证的替代标志物和确定的预防性治疗。以来 艾滋病毒感染者的吸烟率是一般人群的三到四倍。 艾滋病毒感染的非洲裔美国人（AAs）占艾滋病毒感染者的42%， 美国，迫切需要研究吸烟对患有HIV的AA的神经元功能障碍的影响。 线粒体功能障碍是各种神经系统疾病的标志。线粒体损伤和 线粒体DNA（mtDNA）含量的减少通常发生在神经元变性之前。我们的长期目标 是开发一种临床适用的，非侵入性的测试，以监测吸烟对神经元的影响， HIV阳性个体的mtDNA含量。这项测试将允许临床医生监测神经元mtDNA的变化 在艾滋病毒感染的吸烟者的抗逆转录病毒治疗，以促进早期医疗干预的内容。我们的整体 目的探讨吸烟对大鼠海马神经元细胞外基质线粒体DNA含量的影响。 根据吸烟和HIV状态，从AA的外周血中分离出囊泡（NEV）。我们的中央 一种假说是，吸烟加剧了病毒抑制的HIV阳性患者神经元mtDNA损伤， AA，导致外周血NEV中mtDNA释放增加。我们的理论基础是， NEVs是一种新的非侵入性生物标志物，用于早期检测神经元中的mtDNA损伤。我们的具体 目的是：1）定量从AA非吸烟者外周血中分离的NEV的mtDNA含量， 包括病毒抑制的HIV阳性和HIV阴性受试者; 2）比较 从病毒抑制的HIV阳性AA吸烟者外周血中分离的NEV， 从病毒抑制的HIV阳性非吸烟者以及HIV阴性吸烟者中分离的NEV中的含量 和不吸烟者。在目标#1中，我们将测量从外周血中分离的NEV中的mtDNA含量， 使用囊内DNA通过实时定量PCR（qPCR）检测HIV阴性和HIV阳性非吸烟者 从NEV中提取。在目标2中，我们将分析NEV中mtDNA含量的定量变化， 反映了与吸烟和ART相关的mtDNA损伤。我们的建议是创新的，因为它 建议使用外周血NEV中的mtDNA含量作为监测的新替代生物标志物 神经元mtDNA损伤我们从这项研究中的发现将是重要的，他们将建立线粒体DNA NEV中的含量作为实时监测神经元功能障碍的新型非侵入性生物标志物 与HIV阳性个体ART和吸烟者的mtDNA损伤相关。