Opiate abuse and sex steroids influence neuroAIDS pathology

阿片类药物滥用和性类固醇影响神经艾滋病病理学

• 批准号: 9495796
• 负责人: Jason Richard Paris
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF MISSISSIPPI
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9495796
• 关键词: Acquired Immunodeficiency Syndrome; Address; Affect; Affective; Aging; Allopregnanolone; Animal Model; Animals; Anxiety; Astrocytes; Attenuated; Award; Behavior; Behavioral; Behavioral Genetics; Brain Pathology; Calcium; Cells; Chronic; Clinical Trials; Coculture Techniques; Cognition; Cognition Disorders; Cognitive; Contracts; Corpus striatum structure; Data; Dendrites; Dendritic Spines; Development; Dose; Drug usage; Endocrine; Enhancers; Enzymes; Estradiol; Estrogen Receptor alpha; Estrogens; Etiology; Female; Finasteride; Formestane; Gender; General Population; Gonadal Steroid Hormones; Grant; HIV; HIV Envelope Protein gp120; HIV-1; Hormonal; Hormones; Human; Individual; Infection; Injecting drug user; Intractable Pain; Ions; MK801; Maintenance; Mediating; Membrane Potentials; Menopause; Mentors; Microglia; Mitochondria; Molecular; Moods; Morphine; Morphology; Motor; Mus; N-Methyl-D-Aspartate Receptors; National Institute of Drug Abuse; Neurons; Nitrites; Nuclear; Opiates; Opioid; Outcome Measure; Pathogenicity; Pathology; Pathway interactions; Periodicity; Pharmacodynamics; Pharmacology; Phase; Physiological; Placebos; Pregnanes; Process; Production; Progesterone; Progestins; Proteins; Reactive Oxygen Species; Recording of previous events; Regulation; Research; Risk; Sex Bias; Source; Steroids; Tamoxifen; Techniques; Testing; Therapeutic Uses; Time; Tissues; Toxic Actions; Transgenic Mice; Transgenic Organisms; Woman; age group; aged; associated symptom; biocytin; chemokine; connective tissue-activating peptide; cytokine; density; experience; experimental study; fetal; fluorescence imaging; gender difference; genetic approach; hormone therapy; immunocytochemistry; in vivo; inhibitor/antagonist; innovation; male; men; mitochondrial dysfunction; mitochondrial membrane; mouse model; mu opioid receptors; nerve stem cell; neurobehavioral; neuronal cell body; neuropathology; neuropsychiatry; neurosteroids; neurotoxicity; novel; opioid abuse; opioid use; pre-clinical; prescription opioid; programs; protective effect; receptor; relating to nervous system; sex; steroid hormone; translational approach; transmission process

Individuals with human immunodeficiency virus (HIV) experience brain pathology associated with greater motor/mood/cognitive disorders (collectively termed "neuroAIDS") than the general population and these effects are exacerbated among opiate abusers. Moreover, gender differences exist with opiate-abusing women at greater risk to contract HIV compared to opiate-abusing men; yet, some women may experience lesser neuroAIDS symptomology than men once infection occurs. We recapitulated these effects in mice and have begun to elucidate the sex steroid hormones that interact with opiates to confer protection to neuroAIDS. The present proposal will begin to reveal the effects and mechanisms of sex steroid interactions with opiates and HIV in a translational approach that utilizes cultured murine and human neural cells, as well as transgenic whole-animal murine models. This Pathway to Independence Award (K99/R00) will begin to discern the cellular/molecular mechanisms by which sex steroid milieu may influence opiate/HIV interactions for neuroAIDS pathology. In the R00 independent phase of this grant, we will examine the protective effects of central steroid formation on opiate/HIV protein interactions in vivo using whole-animal models that conditionally-express central HIV-1 Tat (Aim 3a) or constitutively-express central gp120 (Aim 3b). Neurosteroid formation will be pharmacologically inhibited or facilitated in male and female mice and subjects will be assessed for neuroAIDS-like motor, affective, and cognitive behavior. Striatal tissue will be assessed for viable/degenerating neurons, astrocytes, and microglia via immunocytochemistry and viable striatal medium spiny neurons (MSNs) will be assessed for sublethal changes in morphology. The influence of pharmacodynamic targets that overlap between steroids, opiates, X4/R5-tropic HIV, and HIV proteins (Tat or gp120) will be examined in murine neural co-cultures obtained mice and differentiated fetal neural progenitor cells (Aim 3c). Cultures will be pretreated with pharmacological antagonists to NMDA receptors, mGluR1, estrogen receptors (alpha and beta), and mu opioid receptors. As in the prior K99 project (Aims 1-2), cultures will be assessed for MSN morphology, intracellular ion concentrations ([Ca2+]i and [Na+]i) within MSN soma and dendrites, and mitochondrial membrane potential (via JC-10 fluorescence imaging). Supernatants will be assessed for production of cytokines/chemokines, reactive oxygen species, and nitrites. This proposal enables the development of an innovative and independent research program.

人类免疫缺陷病毒 (HIV) 感染者的脑部病变与更大的相关性有关。 运动/情绪/认知障碍（统称为“神经艾滋病”）高于一般人群，并且这些 阿片类药物滥用者的影响更为严重。此外，滥用阿片类药物的女性也存在性别差异 与滥用阿片类药物的男性相比，感染艾滋病毒的风险更大；然而，一些女性可能会经历较少的 一旦感染发生，神经艾滋病的症状就多于男性。我们在小鼠身上重现了这些效果，并发现 开始阐明性类固醇激素与阿片类药物相互作用，为神经艾滋病提供保护。这 目前的提案将开始揭示性类固醇与阿片类药物相互作用的影响和机制 利用培养的小鼠和人类神经细胞以及转基因细胞的转化方法中的艾滋病毒 全动物小鼠模型。该独立之路奖 (K99/R00) 将开始辨别 性类固醇环境可能影响阿片类药物/艾滋病毒相互作用的细胞/分子机制 神经艾滋病病理学。在本次拨款的 R00 独立阶段，我们将检查以下项目的保护作用： 使用整体动物模型研究阿片/HIV蛋白体内相互作用的中心类固醇形成 条件表达中心 HIV-1 Tat（目标 3a）或组成型表达中心 gp120（目标 3b）。神经类固醇 雄性和雌性小鼠中的形成将受到药理抑制或促进，并且受试者将 评估神经艾滋病样运动、情感和认知行为。将评估纹状体组织 通过免疫细胞化学和活纹状体培养基获得活/退化神经元、星形胶质细胞和小胶质细胞 将评估多棘神经元（MSN）的形态学亚致死变化。的影响 类固醇、阿片类药物、X4/R5-tropic HIV 和 HIV 蛋白（Tat 或 gp120）将在小鼠神经共培养物中检查获得的小鼠和分化的胎儿神经祖细胞 细胞（目标 3c）。培养物将用 NMDA 受体、mGluR1、 雌激素受体（α 和 β）和 mu 阿片受体。与之前的 K99 项目（目标 1-2）一样，文化 将评估 MSN 体细胞内的 MSN 形态、细胞内离子浓度（[Ca2+]i 和 [Na+]i） 树突和线粒体膜电位（通过 JC-10 荧光成像）。上清液将是 评估细胞因子/趋化因子、活性氧和亚硝酸盐的产生。该提案使 开发创新和独立的研究计划。