Interaction of HIV-Tat and methamphetamine: Role of ion channels and epigenetics

HIV-Tat 和甲基苯丙胺的相互作用：离子通道和表观遗传学的作用

• 批准号: 8601065
• 负责人: Marilou A. Andres
• 金额: $ 13.67万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8601065
• 关键词: AIDS Dementia Complex; Action Potentials; Affect; Alleles; Alzheimer' s Disease; Apolipoprotein E; Brain; Brain Injuries; Calcium; Cardiac; Cells; Cerebrospinal Fluid; Cessation of life; Cholesterol; Clinical Research; Cognitive; CpG Islands; DNA; DNA Methylation; Data; Disease; Disease Progression; Drug usage; Epigenetic Process; Equilibrium; Exhibits; Exons; Future; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Goals; HIV; HIV Infections; HIV tat Protein; Head; Homeostasis; Human; Illicit Drugs; Impairment; Incidence; Individual; Interphase Cell; Ion Channel; Lead; Learning; Lipids; Measures; Mediating; Membrane Potentials; Methamphetamine; Methylation; Monitor; Natural regeneration; Nature; Neurites; Neurologic; Neuronal Injury; Neurons; Outcome; Pathogenesis; Patients; Pattern; Performance; Peripheral; Pharmaceutical Preparations; Physiological; Play; Potassium Channel; Protein Isoforms; Proteins; Recycling; Research; Research Personnel; Rest; Risk Factors; Role; Severities; Staging; Testing; Toxic effect; Viral Proteins; Work; base; brain tissue; career; central nervous system injury; drug of abuse; electrical property; genetic risk factor; neuron loss; neurotoxic; patch clamp; performance tests; promoter; public health relevance; relating to nervous system; repaired; response; voltage clamp

DESCRIPTION (provided by applicant): Methamphetamine (METH) is an illicit drug commonly used (and abused) by HIV-infected patients. METH exacerbates HIV-related neurological impairments and brain injury. Clinical studies show that HIV and METH have additive effects on neuronal loss and glial activation compared to either HIV or METH insult alone. Recent findings also demonstrate that APOE4 suppress K+ channel activities while METH elicits bursts of action potentials. APOE4, a product of the APOE ?4 allele, is a risk factor for Alzheimer's Disease. The APOE ?4 allele is also implicated in accelerating the progression of HIV Disease. To date, no study has been conducted to investigate how APOE gene polymorphisms affect the nature of CNS injury in the presence of HIV infection and in combination with METH use. Our long-term goal is to elucidate the role of genetic variations and their effects on the brain in the presence f both HIV infection and the drugs commonly abused by these individuals. The objective of this proposed work is to better understand the interactions between APOE protein, HIV-Tat protein and METH in promoting neuronal injury via disruption of electrical and physiological responses. Our overall hypothesis is that differential expression of APOE isoforms will determine the severity of neural damage induced by HIV-Tat (during HIV infection) and by METH by directly influencing electrical activities and consequently affecting intracellular calcium (Ca2+) homeostasis. In Aim 1, we will characterize the DNA methylation patterns of post-mortem brain tissues of HIV+ and HIV + METH individuals who are carriers (?4+) and non-carriers of the APOE ?4 allele (?4-). We will determine how methamphetamine use changes the DNA methylation patterns of the APOE promoter and exon 4 CpG islands in HIV-individuals who are ?4+ and ?4-. In Aim 2, we will examine neuronal responses to Tat and METH in the presence of APOE3 and APOE4 proteins by monitoring neurons' electrical activities (Ca2+, Na+, and K+ currents and resting membrane potentials). To achieve this, we will measure macroscopic Ca2+, Na+, and K+ currents and membrane potentials changes using whole-cell patch clamp recordings (in voltage clamp and current clamp modes). These recordings will be done in different APOE isoform backgrounds with and without METH. The data from this research will form the basis of a future R01-level proposal for this New Investigator applicant who is at the early stage of her research career.

描述（由申请人提供）：甲基苯丙胺（冰毒）是艾滋病毒感染者常用（和滥用）的非法药物。冰毒会加重hiv相关的神经损伤和脑损伤。临床研究表明，与单独感染HIV或冰毒相比，HIV和冰毒对神经元丧失和神经胶质活化具有累加性影响。最近的研究结果还表明，APOE4抑制K+通道活动，而甲基安非他明引起动作电位的爆发。APOE4, APOE ？4个等位基因，是阿尔茨海默病的危险因素。APOE ？4等位基因也与加速HIV疾病的进展有关。迄今为止，还没有研究调查APOE基因多态性如何影响艾滋病毒感染和冰毒联合使用时中枢神经系统损伤的性质。我们的长期目标是阐明基因变异的作用及其在艾滋病毒感染和这些个体通常滥用药物的情况下对大脑的影响。这项工作的目的是为了更好地了解APOE蛋白、HIV-Tat蛋白和甲基安非他明之间的相互作用，通过破坏电和生理反应来促进神经元损伤。我们的总体假设是，APOE亚型的差异表达将决定HIV- tat（在HIV感染期间）和甲基安非他明诱导的神经损伤的严重程度，直接影响电活动，从而影响细胞内钙（Ca2+）稳态。在Aim 1中，我们将描述HIV+和HIV+冰毒携带者死后脑组织的DNA甲基化模式(？4+)和非APOE携带者？4等位基因（?4-）。我们将确定使用甲基苯丙胺如何改变艾滋病毒携带者的APOE启动子和外显子4 CpG岛的DNA甲基化模式。4+和？4-在Aim 2中，我们将通过监测神经元的电活动（Ca2+, Na+， K+电流和静息膜电位）来检测APOE3和APOE4蛋白存在时神经元对Tat和METH的反应。为了实现这一目标，我们将使用全细胞膜片钳记录（在电压钳和电流钳模式下）测量宏观Ca2+， Na+和K+电流和膜电位的变化。这些记录将在不同的APOE异构体背景下进行，有和没有冰毒。这项研究的数据将构成该新研究者申请人未来r01级提案的基础，该申请人处于其研究生涯的早期阶段。