Cysteinyl Leukotrienes in HIV Brain Injury

半胱氨酰白三烯在 HIV 脑损伤中的作用

• 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
• 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

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：感染HIV-1可导致痴呆症，目前尚无治疗方法。来自我们和其他实验室的实验证据有力地表明，HIV-1感染和对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和神经发生障碍。我们最近观察到，有丝分裂原激活的蛋白激酶(MAPK)p38在诱导HIV/gp120毒性的M？/小胶质细胞和M？毒素诱导的神经元中都是必需的。在后续研究中，我们发现p38？MAPK通过特异性siRNAs下调M的半胱氨酰白三烯合成酶(LTC4S)。我们还发现，阻断半胱氨酰-白三烯受体1(CysLTR1)可以保护大脑皮层神经元免受gp120刺激或HIV感染M的毒性。因此，我们建议在体内研究LTC4S的基因缺失或CysLTR1的阻断对HIV-1或其包膜gp120引起的脑损伤的影响。我们假设，消除CysLT的产生或抑制CysLTR1可以防止HIV和gp120诱导的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。其具体目的是：(1)在体内确定半胱氨酰白三烯合成酶(LTC4S)基因缺失是否能防止HIV/gp120转基因小鼠的神经元损伤和行为障碍。(2)在体内研究药物抑制CysLTR1是否能减轻HIV/gp120转基因小鼠的神经元损伤。(3)在体外评价阻断CysLTR1或敲除LTC4S基因(KO)如何在HIV诱导的巨噬细胞毒素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)杂交，孟鲁司特将用于阻断CysLTR1(SA2)。将分别比较LTC4SKO与野生型以及孟鲁司特与赋形剂治疗的HIV/gp120转基因小鼠SA1和2的基于记忆和认知的行为表现、神经元损伤和胶质细胞激活。对于SA3，除了LTC4SKO和暴露于HIV感染和未感染的原代人M的条件培养液中的野生型大脑皮层神经元和星形胶质细胞外，我们还将使用CysLTR1的药物抑制。所有三个SA都将使用基于多维质谱仪的鸟枪脂组学来分析CysLts，并与大脑不同部分(SA1和2)以及神经元、星形胶质细胞和M？(SA3)中的其他细胞脂和介体进行比较。对于所有三种SAS，神经元的损伤和死亡将在神经元细胞和突触标记的免疫标记和核DNA Hoechst染色后用去卷积显微镜进行分析。这三个特定的目的都将检验这样一个前提，即删除CysLT的产生或阻断CysLTR1可以防止炎症和损伤过程，有利于神经保护机制，如p38MAPK和Caspase 3活性降低，Akt，活性增加。我们还将在体内评估LTC4SKO和CysLTR1阻断是否可以保护记忆和认知，并在HIV/gp120存在的情况下改善胶质细胞增生。