Optical electrophysiology of human primary neurons: role of KCC2 in hyperexcitability induced by HIV-1, Tat, and gp120 and morphine exposure
人类原代神经元的光学电生理学:KCC2 在 HIV-1、Tat、gp120 和吗啡暴露诱导的过度兴奋中的作用
基本信息
- 批准号:9623656
- 负责人:
- 金额:$ 3.71万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2018
- 资助国家:美国
- 起止时间:2018-07-10 至 2020-01-09
- 项目状态:已结题
- 来源:
- 关键词:3-DimensionalAIDS Dementia ComplexAcuteAddressAffectAminobutyric AcidsAnimal ModelAstrocytesBrainBrain-Derived Neurotrophic FactorCCR5 geneCXCR4 ReceptorsCalciumCellsChloride IonChronicComorbidityDataDiseaseDrug usageElectrophysiology (science)EnhancersEnvironmentEquilibriumExcitatory NeurotoxinsExposure toFunctional disorderGlutamatesHIVHIV Envelope Protein gp120HIV InfectionsHIV-1HIV-associated neurocognitive disorderHumanImageIn VitroIndividualInfectionInflammation MediatorsInflammatoryInterventionLeadLightMaintenanceMeasuresMediatingMediator of activation proteinMicrogliaModelingMorphineN-Methyl-D-Aspartate ReceptorsNeuronal DysfunctionNeuronsOpioidOpioid ReceptorOpticsOutcomePathologyPharmacologyPhysiologicalPlayPrevalencePropertyProteinsProtocols documentationReceptor ActivationReceptor Protein-Tyrosine KinasesResearchRoleSignal PathwaySignal TransductionSymptomsSynapsesSystemT-LymphocyteTechniquesTestingWithdrawalWorkantiretroviral therapyastrocyte progenitorcalcium indicatorcell typechemokine receptorchloride-cotransporter potassiumexposed human populationfetalgamma-Aminobutyric Acidgamma-Chemokineshuman modelinhibitor/antagonistinnovationmacrophagemu opioid receptorsnerve stem cellnervous system disorderneuronal excitabilityneurotoxicnew therapeutic targetnovelopioid useopioid use disorderoptogeneticsreceptorrecruitreduce symptomsrelating to nervous systemresponsetoolvoltage
项目摘要
PROJECT SUMMARY
The introduction of combined antiretroviral therapy has decreased the prevalence of HIV-associated dementia,
but prevalence of milder HIV-associated neurocognitive disorders (HAND) has increased. The CNS is highly
vulnerable to insult from HIV-1 via release of neurotoxic HIV proteins, inflammatory factors, and excitotoxins by
infected astrocytes and microglia. Opiate use is often comorbid with HIV-1 infection and there is evidence for
enhanced HAND symptomology in users, likely due to convergent cell signaling pathways resulting in increased
neuronal dysfunction. Despite intense research the interactions between HIV-1 and opiates that lead to neuronal
damage remain elusive, and most of this work is conducted in animal models. Since HIV-1 is a human-specific
disease, it is paramount to validate and expand upon research findings in nonhuman models using human
models. To this end, we have differentiated human neural progenitors to develop two primary human CNS culture
models: i) a dissociated system containing glutamatergic and GABAergic neurons; and ii) a 3-dimensional brain
aggregate (BrAgg) model that contains all major CNS cell types. Using non-invasive electrophysiological
techniques including genetically encoded voltage and Ca2+ indicators (GEVI/GECI), and optogenetics we will
examine the effects of infectious HIV-1, Tat, and gp120 ± morphine on primary human neuron function. By
studying HIV-1 ± morphine effects on neuronal excitability we have uncovered a novel therapeutic target. K-Cl
cotransporter 2 (KCC2) maintains low neuronal [Cl-]i necessary for -aminobutyric acid type A receptor
(GABAAR)-mediated inhibition. Loss of KCC2 activity resulting in loss of GABAAR hyperpolarization, and even
GABA-induced depolarization, has been implicated in a variety of neurological disorders. This project tests the
hypothesis that HIV-1 and morphine-induced signaling converge to reduce neuronal KCC2 activity leading to
electrophysiological and synaptic excitatory-inhibitory imbalance in primary human neurons. Our data have
demonstrated significant loss of KCC2 and subsequent shift from hyper- to depolarization in response to GABA
in HIV-1, Tat, and gp120 ± morphine-exposed human neurons. Pharmacological intervention to raise KCC2
levels has reversed these effects. Aim 1 will utilize GEVI and GECI to determine functional responses to
infectious HIV-1, Tat, and gp120 (R5-, X4-, and dual-tropic) exposure on dissociated human neurons and
examine modulation of KCC2 and relevant upstream regulators (e.g. NMDAR, CCR5, CXCR4, opioid receptors)
to rescue noted deficits. Aim 2 will utilize stable BrAgg which contain all major CNS cell types and can be actively
infected by HIV-1. Tools and measures described in Aim 1 will be used to examine neuronal function in a chronic,
HIV-1-infected environment and elucidate differences between chronic, acute, and withdrawal morphine
exposure paradigms. These studies use innovative models and techniques to determine effects of live HIV-1,
Tat, and gp120 ± morphine on primary human neuron function and identify novel points of HIV-1 and morphine
signaling convergence and targets for intervention to alleviate the symptoms of HAND ± comorbid opiate use.
项目概要
联合抗逆转录病毒疗法的引入降低了艾滋病毒相关痴呆症的患病率,
但较轻微的 HIV 相关神经认知障碍 (HAND) 的患病率却大幅增加。
通过释放神经毒性 HIV 蛋白、炎症因子和兴奋毒素,容易受到 HIV-1 的损害
受感染的星形胶质细胞和小胶质细胞使用阿片类药物通常与 HIV-1 感染并存,并且有证据表明。
用户的 HAND 症状增强,可能是由于细胞信号通路趋同导致增加
尽管进行了大量研究,但 HIV-1 和阿片类药物之间的相互作用会导致神经元功能障碍。
损害仍然难以捉摸,而且大部分工作都是在动物模型中进行的,因为 HIV-1 是人类特异性的。
疾病,至关重要的是使用人类在非人类模型中验证和扩展研究结果
为此,我们分化了人类神经祖细胞来开发两种主要的人类中枢神经系统培养物。
模型:i) 包含谷氨酸能和 GABA 能神经元的分离系统;ii) 3 维大脑;
包含所有主要 CNS 细胞类型的聚合 (BrAgg) 模型,使用非侵入性电生理学。
技术,包括基因编码电压和 Ca2+ 指示剂 (GEVI/GECI) 以及光遗传学,我们将
检查传染性 HIV-1、Tat 和 gp120 ± 吗啡对人类初级神经元功能的影响。
通过研究 HIV-1±吗啡对神经元兴奋性的影响,我们发现了一个新的治疗靶点。
协同转运蛋白 2 (KCC2) 维持 γ-氨基丁酸 A 型受体所需的低神经元 [Cl-]i
(GABAAR) 介导的抑制导致 KCC2 活性丧失,甚至导致 GABAAR 超极化丧失。
GABA 诱导的去极化与多种神经系统疾病有关。
假设 HIV-1 和吗啡诱导的信号传导会聚合以减少神经元 KCC2 活性,从而导致
我们的数据显示,人类原代神经元的电生理学和突触兴奋抑制失衡。
KCC2 显着丧失,随后响应 GABA 从超极化转变为去极化
HIV-1、Tat 和 gp120 ± 吗啡暴露的人类神经元中的药物干预可提高 KCC2。
目标 1 将利用 GEVI 和 GECI 来确定对这些影响的功能反应。
传染性 HIV-1、Tat 和 gp120(R5-、X4- 和双向)暴露于分离的人类神经元,
检查 KCC2 和相关上游调节因子(例如 NMDAR、CCR5、CXCR4、阿片受体)的调节
目标 2 将利用包含所有主要中枢神经系统细胞类型并且可以主动激活的稳定 BrAgg。
目标 1 中描述的工具和措施将用于检查慢性、
HIV-1 感染环境并阐明慢性、急性和戒断吗啡之间的差异
这些研究使用创新模型和技术来确定活 HIV-1 的影响,
Tat 和 gp120 ± 吗啡对人类初级神经元功能的影响以及 HIV-1 和吗啡的新识别点
信号趋同和干预目标,以减轻 HAND ± 合并阿片类药物使用的症状。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Aaron Joseph Barbour其他文献
Aaron Joseph Barbour的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Aaron Joseph Barbour', 18)}}的其他基金
Optical electrophysiology of human primary neurons: role of KCC2 in hyperexcitability induced by HIV-1, Tat, and gp120 and morphine exposure
人类原代神经元的光学电生理学:KCC2 在 HIV-1、Tat、gp120 和吗啡暴露诱导的过度兴奋中的作用
- 批准号:
9754573 - 财政年份:2018
- 资助金额:
$ 3.71万 - 项目类别:
相似国自然基金
化学小分子对于G蛋白偶联受体介导的蛋白-蛋白相互作用影响的研究
- 批准号:90713047
- 批准年份:2007
- 资助金额:300.0 万元
- 项目类别:重大研究计划
相似海外基金
Novel Radial Diffusion-Weighted MR Spectroscopic Imaging of HIV: Biomarker Detection Using Functional Imaging and Neurocognitive Correlates
HIV 的新型径向扩散加权 MR 光谱成像:使用功能成像和神经认知相关性进行生物标志物检测
- 批准号:
10256718 - 财政年份:2020
- 资助金额:
$ 3.71万 - 项目类别:
Identify New Determinants to Target Macrophage-Tropic Viruses and Prevent HIV-1 Brain Infection in NeuroAIDS
确定针对巨噬细胞嗜性病毒并预防神经艾滋病中 HIV-1 脑部感染的新决定因素
- 批准号:
10370309 - 财政年份:2018
- 资助金额:
$ 3.71万 - 项目类别:
Identify New Determinants to Target Macrophage-Tropic Viruses and Prevent HIV-1 Brain Infection in NeuroAIDS
确定针对巨噬细胞嗜性病毒并预防神经艾滋病中 HIV-1 脑部感染的新决定因素
- 批准号:
9893028 - 财政年份:2018
- 资助金额:
$ 3.71万 - 项目类别:
Identify New Determinants to Target Macrophage-Tropic Viruses and Prevent HIV-1 Brain Infection in NeuroAIDS
确定针对巨噬细胞嗜性病毒并预防神经艾滋病中 HIV-1 脑部感染的新决定因素
- 批准号:
9618409 - 财政年份:2018
- 资助金额:
$ 3.71万 - 项目类别:
CD8 T cell mediated disruption of Blood Brain Barrier Tight Junctions
CD8 T 细胞介导的血脑屏障紧密连接破坏
- 批准号:
9392836 - 财政年份:2017
- 资助金额:
$ 3.71万 - 项目类别: