Chloride channel-dependent mechanisms of opiate and HIV-induced synaptodendritic injury

阿片类药物和 HIV 诱导的突触树突损伤的氯离子通道依赖性机制

• 批准号: 10548312
• 负责人: Kurt F Hauser
• 金额: $ 19.41万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10548312
• 关键词: AMPA Receptors; Acute; Affect; Afferent Neurons; Astrocytes; Behavioral; Blood - brain barrier anatomy; Brain; Buffers; CLC-1 channel; Cell Line; Cells; Chloride Channels; Chlorides; Complex; Corpus striatum structure; Coupling; Data; Disinhibition; Dopamine D1 Receptor; Drug Targeting; Electrophysiology (science); Equilibrium; Exposure to; Functional disorder; Genetic; Glutamates; Glycine Receptors; HIV; HIV Envelope Protein gp120; HIV-1; HIV-associated neurocognitive disorder; Homeostasis; Immunosuppression; In Vitro; Injury; Interneurons; Intervention; Ion Channel; Long-Term Effects; Mediating; Membrane; Microglia; Modeling; Molecular; Molecular Biology; Morphine; Morphology; N-Methyl-D-Aspartate Receptors; Neuroglia; Neuronal Injury; Neurons; Opioid; Pathology; Peripheral; Persons; Pharmacology; Regulation; Role; Signal Transduction; Site; Slice; Source; Synapses; System; Techniques; Testing; Therapeutic; Time; Transgenic Mice; behavioral outcome; central nervous system injury; chloride-cotransporter potassium; confocal imaging; cytokine; driving force; excitotoxicity; gain of function; gamma-Aminobutyric Acid; genetic approach; in vivo; loss of function; mouse model; nervous system disorder; neuroAIDS; neuroinflammation; neuron loss; neuronal excitability; neurotoxic; novel; opioid abuse; opioid use disorder; postsynaptic; presynaptic; synaptic inhibition; voltage

Opioid use disorder (OUD) in people infected with HIV-1 (PWH) exacerbates the pathobiology of neuroHIV and HIV-associated neurocognitive disorder (HAND) via complex mechanisms involving direct peripheral (e.g., immune suppression) and CNS (e.g., neuroinflammation, blood-brain barrier damage, and neuronal injury) effects. Although glutamatergic excitotoxicity remains the major driving force in HIV-dependent neuronal injury, emerging evidence from our lab indicates that the loss of inhibitory GABAergic control and [Cl−]i homeostasis not only contributes to a loss of inhibition, but also increases the excitatory/neurotoxic effects of glutamate at NMDA and AMPA receptors. This proposal directly tests the assumption that [Cl−]i-dependent net losses of neuronal inhibition will increase excitation and promote excitotoxic. In mature neurons, intracellular Cl− concentration ([Cl−]i) is dynamically regulated by membrane ion channels (GABAA and glycine receptors) and Cl− channels and transporters. By controlling [Cl−]i, these channels and transporters regulate the Cl− equilibrium potential (ECl) and neuronal inhibition and we have already shown the KCC2 loss-of-function in OUD/neuroHIV models leads to [Cl−]i increases, elevated ECl, and excitotoxicity. We discovered a previously unidentified a ClC-1-like voltage-dependent Cl− channel that differentially regulates the excitability of dopamine receptor D1- and D2- expressing (D1 and D2, respectively) striatal medium spiny neurons (MSNs) and is disrupted by opioids and HIV Tat. We hypothesize that opioids and HIV-1 disrupt [Cl−]i, homeostasis via a voltage-gated ClC Cl− channel resulting in synaptodendritic injury. This hypothesis will be tested in the following specific aims and if confirmed will be a conceptual shift in our understanding of how opioids and HIV contribute to excitotoxicity. Aim 1 will identify the mechanisms by which opioids and HIV dysregulate voltage- dependent ClC-1 channels and Cl− homeostasis in vitro. Our preliminary findings suggest that ClC-1 significantly affects voltage-dependent Cl− currents in D1 and D2 MSNs. The extent to which ClC-1 mediates the HIV and opioid-induced pathology will be explored using gain-of-function, loss-of-function strategies, and confirmed in vitro and in vivo/ex vivo using electrophysiologic, pharmacologic, and genetic approaches at the cell and molecular levels. Aim 2. Establish to what extent ClC-1 modulation contributes to the synaptodendritic injury caused by opioids and HIV-induced dysregulation of Cl− homeostasis. We predict that the dysregulation of [Cl−]i homeostasis amplifies synaptodendritic injury in MSNs in OUD/neuroHIV models. Since ClC-1 can be a novel source of activity-dependent Cl− entry, we also predict that opioids and HIV Tat cause neuronal injury via a complex mechanism involving ClC-1 modulation and, thus, synaptic disinhibition. This project will test whether opioids and HIV dysregulate [Cl−]i homeostasis and cause neuronal injury via ClC-1. If ClC-1 is confirmed to be that target, as preliminary evidence indicates, this will be a conceptual shift in our understanding of how opioids and HIV contribute to excitotoxicity.

感染HIV-1（PWH）的人的阿片类药物使用障碍（OUD）加剧了神经hiv的病理学和 通过涉及直接外围的复杂机制与HIV相关的神经认知障碍（手）（例如， 免疫抑制）和中枢神经系统（例如，神经炎症，血脑屏障损伤和神经元损伤） 尽管谷氨酸能兴奋性仍然是HIV依赖性神经元损伤的主要驱动力，但 来自我们实验室的新兴证据表明抑制性GABA能控制和[Cl-] I稳态的丧失 不仅有助于抑制丧失，而且还增加了谷氨酸的兴奋/神经毒性作用 NMDA和AMPA接收器。该建议直接测试了[Cl-] i依赖性的净损失的假设 神经元抑制作用会增加兴奋并促进兴奋性毒性。在成熟的神经元中，细胞内CL- 浓度（[Cl-] i）受膜离子通道（GABAA和甘氨酸受体）和 Cl-通道和转运蛋白。通过控制[Cl-] I，这些通道和转运蛋白调节Cl- 平衡电位（ECL）和神经元抑制作用，我们已经显示了KCC2功能丧失 OUD/NEUROHIV模型导致[Cl-] I增加，ECL升高和兴奋性毒性。我们发现了一个以前的 身份不明的Clc-1样电压依赖性Cl-通道，对多巴胺的刺激性不同 受体D1和D2-表达（D1和D2）纹状体培养基神经元（MSN），IS 被杀菌和HIV TAT破坏。我们假设Opioids和HIV-1破坏[Cl-] I，稳态通过 电压门控的CLC CL-通道导致突触don损伤。该假设将在 遵循特定目标，如果确认将是我们对阿片类药物和艾滋病毒的理解的概念转变 有助于兴奋性。 AIM 1将确定阿片类药物和HIV失调的机制 - 电压 - 体外依赖性CLC-1通道和Cl-稳态。我们的初步发现表明CLC-1 显着影响D1和D2 MSN中电压依赖性的cl液。 CLC-1培养基的程度 艾滋病毒和阿片类药物诱导的病理将使用功能障碍，功能丧失策略和 使用电生理，药理和遗传方法在体外和体内确认 细胞和分子水平。目标2。确定CLC-1调制在多大程度上有助于突触 阿片类药物和HIV诱导的CL Homeostasis失调引起的损伤。我们预测失调 OUD/NEUROHIV模型中MSN中的[Cl-] I稳态放大器突触损伤。因为Clc-1可以是 活性依赖性Cl-进入的新型来源，我们还预测阿片类药物和HIV TAT通过 涉及CLC-1调制的复杂机制，因此是突触抑制作用。这个项目将测试 阿片类药物和HIV失调[CL-] I稳态不体，并通过CLC-1引起神经元损伤。如果Clc-1是 正如初步证据所表明的那样，确认目标是我们的概念转变 了解阿片类药物和艾滋病毒如何促进兴奋性。