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 感染可诱发痴呆症，目前尚无有效治疗方法。我们和其他实验室的实验证据表明，HIV-1 感染和大脑中小胶质细胞和巨噬细胞 (M�) 的神经毒性刺激会引发神经元损伤。我们最近观察到，丝裂原激活蛋白激酶（MAPK）p38在M/小胶质细胞中是诱导HIV/gp120毒性和在神经元中启动细胞凋亡所必需的。在后续研究中，我们发现特定 siRNA 敲低 p38? MAPK 会下调 M� 中的半胱氨酰白三烯合酶 (LTC4S)。我们还发现，阻断半胱氨酰白三烯受体 1 (CysLTR1) 可以保护大脑皮质。因此，我们建议在体内研究如何抵抗 gp120 刺激或 HIV 感染的 M 的毒性。 LTC4S 的基因缺失或 CysLTR1 的阻断会影响 HIV-1 或其包膜 gp120 引起的脑损伤，我们发现消除 CysLT 的产生或抑制 CysLTR1 可以防止 HIV 和 gp120 诱导神经元损伤和行为障碍。寻找新的针对HIV感染脑损伤的保护策略，具体目标（SA）是：（1）确定体内半胱氨酰白三烯合酶是否存在基因缺失。 (LTC4S) 可预防 HIV/gp120 转基因小鼠模型中的神经元损伤和行为障碍 (2) 体内研究 CysLTR1 的药物抑制是否可改善 HIV/gp120 转基因小鼠模型中的神经元损伤 (3) 体外评估如何改善。阻断 CysLTR1 或敲除 (KO) LTC4S 可使神经元在 HIV 诱导的表达巨噬细胞毒素的情况下存活。 HIV/gp120 将与 LTC4SKO 动物 (SA1) 杂交，孟鲁司特将用于阻断 CysLTR1 (SA2)，并将 LTC4SKO 与野生型和基于记忆和认知的行为表现、神经损伤和神经胶质细胞激活进行比较。孟鲁司特与媒介物处理的 HIV/gp120 转基因小鼠分别针对 SA1 和 2，对于 SA3，我们将使用药物抑制。 CysLTR1 除了 LTC4SKO 以及暴露于 HIV 感染和未感染的条件培养基的野生型脑皮质神经元和星形胶质细胞外，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 与其他药物的比较。大脑不同部分（SA1 和 2）以及神经元、星形胶质细胞和 M�（SA3）中的细胞脂质和介质。在对神经元细胞和突触标记进行免疫标记并用 Hoechst 染料对核 DNA 进行染色后，将通过反卷积显微镜对神经元损伤和死亡进行分析。所有三个具体目标将测试 CysLT 产生的删除或 CysLTR1 的阻断可预防炎症和损伤过程的前提。有利于神经保护机制，例如 p38 MAPK 和 Caspase 3 的活性降低，以及 Akt 的活性增加，我们还将评估体内是否存在。 LTC4SKO 和 CysLTR1 阻断可以在 HIV/gp120 存在的情况下保留记忆和认知并改善神经胶质增生。