Cysteinyl Leukotrienes in HIV Brain Injury

半胱氨酰白三烯在 HIV 脑损伤中的作用

基本信息

批准号：
8790362
负责人：
MARCUS KAUL
金额：
$ 48.75万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2020-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8790362
关键词：
Ablation Acquired Immunodeficiency Syndrome Affect Allergic rhinitis Anabolism Animals Apoptosis Astrocytes Behavioral Brain Brain Injuries Brain Part Breeding CXCR4 gene Cellular Stress Cessation of life Chemokine (C-C Motif) Receptor 5 Cognition Conditioned Culture Media DNA Dementia Development Disease Dyes Event Extrinsic asthma Future Genetic Gliosis Glycoproteins HIV HIV Envelope Protein gp120 HIV-1 HIV-associated neurocognitive disorder Human Impairment In Vitro Individual Infection Inflammation Inflammation Mediators Inflammatory Injury Knock-out Laboratories Leukotriene A4 Link Lipids MAPK14 gene MAPK8 gene Macrophage Activation Mass Spectrum Analysis Mediating Medical Research Memory Methodology Microglia Microscopy Mitogen Activated Protein Kinase 1 Mitogen-Activated Protein Kinases Molecular Nerve Degeneration Neuraxis Neuroglia Neuronal Injury Neurons Nuclear Pathway interactions Performance Phenotype Process Production Public Health Research Research Institute Rodent Role Shotguns Signal Pathway Signal Transduction Staining method Stains Synapses Techniques Testing Therapeutic Toxic effect Toxin Transgenic Mice Virus Virus Diseases Work astrogliosis base behavioral impairment biological systems caspase-3 cysteinyl leukotriene receptor cysteinyl-leukotriene env Gene Products extracellular signal-regulated kinase 3 follow-up human MAPK14 protein immune activation improved in vitro Model in vivo in vivo Model inhibitor/antagonist lipid mediator macrophage mitogen-activated protein kinase p38 montelukast mouse model neurocognitive disorder neurogenesis neuron apoptosis neuronal survival neuroprotection neurotoxic neurotoxicity novel prevent public health relevance receptor research study therapeutic target therapy development uptake

项目摘要

DESCRIPTION: Infection with HIV-1 can induce dementia for which a treatment is currently not available. Experimental evidence from our and other laboratories strongly suggests that HIV-1 infection and neurotoxic stimulation of microglia and macrophages (M¿) in the brain trigger neuronal damage and impairment of neurogenesis. We recently observed that mitogen-activated protein kinase (MAPK) p38 was required in both M¿/microglia for induction of HIV/gp120 toxicity and in neurons for initiation of apoptosis by M¿ toxins. In follow-up studies we found that knockdown of p38? MAPK by specific siRNAs down-regulated cysteinyl leukotriene synthase (LTC4S) in M¿. We also discovered that blockade of the cysteinyl-leukotriene receptor 1 (CysLTR1) protected cerebrocortical neurons against toxicity of gp120-stimulated or HIV-infected M¿. Therefore, we propose to study in vivo how genetic deletion of LTC4S or blockade of CysLTR1 affects brain injury caused by HIV-1 or its envelope gp120. We hypothesize that ablation of CysLT production or CysLTR1 inhibition can prevent HIV and gp120 from inducing neuronal injury and behavioral impairment. The long-term objectives are to find new protective strategies against brain injury by HIV infection. The specific aims (SA) are: (1) To determine in vivo whether genetic deletion of cysteinyl-leukotriene synthase (LTC4S) prevents neuronal injury and behavioral impairment in a HIV/gp120 transgenic mouse model. (2) To investigate in vivo whether pharmacological inhibition of CysLTR1 ameliorates neuronal damage in a HIV/gp120 transgenic mouse model. (3) To assess in vitro how the blockade of CysLTR1 or knockout (KO) of LTC4S enables neuronal survival in the presence of HIV-induced macrophage toxins. Transgenic mice expressing HIV/gp120 will be cross-bred with LTC4SKO animals (SA1) and Montelukast will be used to block CysLTR1 (SA2). Memory and cognition-based behavioral performance, neuronal injury and glial cell activation will be compared in LTC4SKO versus wild-type and Montelukast- versus vehicle-treated HIV/gp120-transgenic mice for SA1 and 2, respectively. For SA3, we will use pharmacological inhibition of CysLTR1 besides LTC4SKO and wild-type cerebrocortical neurons and astrocytes exposed to conditioned media of HIV-infected and un-infected, primary human M¿. All three SAs will employ multi-dimensional mass spectrometry-based shotgun lipidomics to profile CysLTs in comparison to other cellular lipids and mediators in different parts of the brain (SA1 and 2) and neurons, astrocytes and M¿ (SA3). For all three SAs neuronal injury and death will be analyzed by deconvolution microscopy after immunolabeling for neuronal cellular and synaptic markers and staining of nuclear DNA by Hoechst dye. All three Specific Aims will test the premise that deletion of CysLT production or blockade of CysLTR1 prevents inflammatory and injurious processes in favor of neuroprotective mechanisms, such as reduced activity of p38 MAPK and Caspase 3, and increased activity of Akt,. We will also assess in vivo whether LTC4SKO and CysLTR1 blockade can preserve memory and cognition and ameliorate gliosis in the presence of HIV/gp120.

描述：用HIV-1感染可以诱导目前尚无治疗的痴呆。来自我们和其他实验室的实验证据强烈表明HIV-1感染和神经毒性刺激小胶质细胞和巨噬细胞（M。）在脑部触发神经元损伤和神经发生损害。我们最近观察到，在M o型 /小胶质细胞中，需要有丝分裂原激活的蛋白激酶（MAPK）p38，以诱导HIV /GP120毒性和神经元在M毒素中启动凋亡的神经元中。在后续研究中，我们发现p38的敲低？ MAPK由特定的siRNA下调的白细胞三烯合酶（LTC4S）。我们还发现，囊这基林 - 白细胞受体1（CYSLTR1）保护了脑皮层神经元免受GP120刺激或HIV感染的M e毒性的毒性。因此，我们建议研究体内LTC4S或CYSLTR1的阻断遗传缺失如何影响由HIV-1或其包膜GP120引起的脑损伤。我们假设消融CYSLT产生或CYSLTR1抑制可以防止HIV和GP120因诱导的神经元损伤和行为障碍而防止HIV和GP120。长期目标是通过HIV感染找到针对脑损伤的新策略。具体目的（SA）是：（1）在体内确定伴半胱烷基列酮二烯合酶（LTC4）的遗传缺失是否可以防止HIV/GP120转基因小鼠模型中的神经元损伤和行为障碍。（2）在体内研究Cysltr1的药理抑制是否可以改善HIV/GP120转基因小鼠模型中的神经元损伤。（3）在体外评估cysltr1或敲除（KO）LTC4S的阻断如何在HIV诱导的巨噬细胞毒素的存在下使神经元存活。表达HIV/GP120的转基因小鼠将与LTC4SKO动物（SA1）交叉，并将使用Monterukast进行阻止Cysltr1（SA2）。 Memory and cognition-based behavioral performance, neuronal injury and glial cell activation will be compared in LTC4SKO versus wild-type and Monterukast- versus vehicle-treated HIV/gp120-transgenic mice for SA1 and 2, For SA3, we will use pharmaceutical inhibition of CysLTR1 besides LTC4SKO and wild-type cerebrocortical neurons and与其他基于质谱的shot弹枪脂肪组学的星形胶质细胞相比，与其他细胞脂质和介体相比，所有三个SA都将采用基于多维的质谱型shot弹式脂肪组学的原发性人，所有三个SA都将使用（SA1和2）和Neurons和Neurons和Neurons，Attrocytes和M.（Sa3）（Sa3）（SA3）（SA3）（SA3）（SA3）（SA3）（SA3）（SA3）（SA3）。对于所有三种SAS神经元损伤和死亡，将通过免疫标记后的反卷积显微镜分析神经元细胞和合成标记物以及Hoechst染料对核DNA的染色。所有三个具体目标都将检验以下前提：删除Cyslt产生或CYSLTR1的阻塞都可以防止炎症和受伤的过程有利于神经保护机制，例如p38 mapk和caspase 3的活性降低，以及Akt的活动增加。我们还将评估在HIV/GP120存在下LTC4SKO和CYSLTR1封锁是否可以保留记忆力，认知和改善神经胶质作用。