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.

患有人类免疫缺陷病毒（HIV）的人会经历与更大的脑病理学 与一般人群相比，运动/情绪/认知障碍（统称为“神经辅助”） 鸦片滥用者之间的影响加剧。此外，性别差异与虐待的妇女存在 与滥用鸦片的男性相比，艾滋病毒的风险更大；但是，有些女性可能会少少 神经辅助症状比一旦发生感染的男性。我们在小鼠中概括了这些影响，并具有 开始阐明与阿片类药物相互作用的性类固醇激素，以赋予神经助剂保护。这 目前的建议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， 使用培养的鼠和人类神经细胞以及转基因的翻译方法中的艾滋病毒 全动物鼠模型。这项获得独立奖的途径（K99/R00）将开始辨别 性类固醇环境可能影响阿片类药物/HIV相互作用的细胞/分子机制 神经辅助病理学。在这笔赠款的R00独立阶段，我们将研究 使用全动物模型在体内的鸦片/HIV蛋白相互作用上的中央类固醇形成 有条件地表达中央HIV-1 TAT（AIM 3A）或组成表达的中央GP120（AIM 3B）。神经类固醇 在雄性和雌性小鼠中，形成将在药理抑制或促进，受试者将是 评估了神经辅助运动，情感和认知行为。将评估纹状体组织 通过免疫细胞化学和可行的纹状体培养基可行/退化神经元，星形胶质细胞和小胶质细胞 将评估多刺神经元（MSN）的形态学变化。影响 类固醇，阿片类药物，X4/R5-热带HIV和HIV蛋白（TAT或TAT或TAT或TAT或 GP120）将在获得小鼠的鼠神经共培养和分化的胎儿神经祖细胞中检查 细胞（AIM 3C）。培养物将通过药理学拮抗剂对NMDA受体MGLUR1， 雌激素受体（Alpha和beta）和MU阿片受体。与先前的K99项目一样（目标1-2），文化 将评估MSN的MSN形态，细胞内离子浓度（[Ca2+] I和[Na+] I） 和树突和线粒体膜电位（通过JC-10荧光成像）。上清液会 评估了细胞因子/趋化因子，活性氧和亚硝酸盐的产生。该建议可以实现 创新和独立研究计划的发展。