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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：
9039665
负责人：
MARCUS KAUL
金额：
$ 43.88万
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/9039665
关键词：
Ablation Acquired Immunodeficiency Syndrome Affect Allergic rhinitis Anabolism Animals Apoptosis Astrocytes Behavioral Brain Brain Injuries Breeding CASP3 gene CCR5 gene CXCR4 gene Cellular Stress 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 MAPK1 gene MAPK14 gene MAPK8 gene Macrophage Activation Mass Spectrum Analysis Mediating Medical Research Memory Methodology Microglia Microscopy Mitogen-Activated Protein Kinases Molecular Nerve Degeneration Neuraxis 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 cysteinyl leukotriene receptor cysteinyl-leukotriene env Gene Products extracellular signal-regulated kinase 3 follow-up glial activation immune activation improved in vitro Model in vivo in vivo Model inhibitor/antagonist knock-down lipid mediator macrophage mitogen-activated protein kinase p38 montelukast mouse model neurocognitive disorder neurogenesis neuron apoptosis neuron loss 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：HIV-1感染可诱发痴呆，目前尚无治疗方法。来自我们和其他实验室的实验证据强烈表明，HIV-1感染和脑中小胶质细胞和巨噬细胞（M？）的神经毒性刺激引发神经元损伤和神经发生障碍。我们最近观察到，丝裂原活化蛋白激酶（MAPK）p38在M β/小胶质细胞中诱导HIV/gp 120毒性和在神经元中启动M β毒素凋亡都是必需的。在后续的研究中，我们发现p38的敲除？MAPK通过特异性siRNA下调半胱氨酰白三烯合酶（LTC 4S）在M。我们还发现，阻断半胱氨酰白三烯受体1（CysLTR 1）可以保护大脑皮层神经元免受gp 120刺激或HIV感染的M β的毒性。因此，我们建议在体内研究LTC 4S的基因缺失或CysLTR 1的阻断如何影响HIV-1或其包膜gp 120引起的脑损伤。我们假设，CysLT生产或CysLTR 1抑制消融可以防止艾滋病毒和gp 120诱导神经元损伤和行为障碍。长期目标是找到新的保护策略，防止艾滋病毒感染造成的脑损伤。具体目的（SA）是：（1）在体内确定半胱氨酰-白三烯合酶（LTC 4S）的遗传缺失是否防止HIV/gp 120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CysLTR 1的药理学抑制是否改善HIV/gp 120转基因小鼠模型中的神经元损伤。(3)在体外评估CysLTR 1阻断或LTC 4S敲除（KO）如何在存在HIV诱导的巨噬细胞毒素的情况下使神经元存活。表达HIV/gp 120的转基因小鼠将与LTC 4SKO动物（SA 1）杂交，孟鲁司特将用于阻断CysLTR 1（SA 2）。将分别在LTC 4SKO与野生型以及孟鲁司特与溶媒处理的HIV/gp 120转基因小鼠中比较SA 1和2的基于记忆和认知的行为表现、神经元损伤和神经胶质细胞活化。对于SA 3，我们将使用药物抑制CysLTR 1，除了LTC 4SKO和野生型皮质神经元和星形胶质细胞暴露于HIV感染和未感染的原代人M β的条件培养基。所有三个SA都将采用基于多维质谱的鸟枪脂质组学来分析CysLT与大脑不同部位（SA 1和2）以及神经元、星形胶质细胞和M β（SA 3）中的其他细胞脂质和介质的关系。对于所有三种SA，在免疫标记神经元细胞和突触标记物并通过Hoechst染料染色核DNA后，通过去卷积显微镜分析神经元损伤和死亡。所有三个特定目的将测试CysLT产生的缺失或CysLTR 1的阻断预防有利于神经保护机制的炎症和损伤过程的前提，例如p38 MAPK和Caspase 3的活性降低和Akt的活性增加。我们还将在体内评估LTC 4SKO和CysLTR 1阻断是否可以在HIV/gp 120存在下保留记忆和认知并改善神经胶质增生。