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: 9114548
• 负责人: Jason Richard Paris
• 金额: $ 11.27万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9114548
• 关键词: Acquired Immunodeficiency Syndrome; Address; Affect; Affective; Animal Model; Animals; Anxiety; Astrocytes; Attenuated; Award; Behavior; Behavioral; Behavioral Genetics; Brain; Brain Pathology; Cell Culture Techniques; Cells; Chronic; Clinical Trials; Coculture Techniques; Cognition; Cognition Disorders; Cognitive; Contracts; Corpus striatum structure; Cultured Cells; Data; Dendrites; Detection; Development; Dose; Drug usage; Endocrine; Enhancers; Enzymes; Estradiol; Estrogens; Etiology; Female; Finasteride; Formestane; Gender; General Population; Glial Fibrillary Acidic Protein; Goals; Gonadal Steroid Hormones; Griess reagent; HIV; HIV Envelope Protein gp120; HIV-1; Health; Homeostasis; Hormonal; Hormones; Human; Image; Imaging Techniques; In Situ Nick-End Labeling; Individual; Infection; Injecting drug user; Intractable Pain; Ions; Life; MK801; Maintenance; Mediating; Membrane; Membrane Potentials; Menopause; Mentors; Microglia; Mitochondria; Molecular; Moods; Morphine; Morphology; Motor; Mus; National Institute of Drug Abuse; Neurons; Nitric Oxide; Nitrites; Opiates; Opioid; Opioid Receptor; Outcome Measure; Paris, France; Pathogenicity; Pathology; Pathway interactions; Pharmacodynamics; Phase; Placebos; Process; Production; Progesterone; Progestins; Proteins; Reactive Oxygen Species; Regulation; Research; Rhodamine 123; Risk; Sex Bias; Source; Steroids; Structure; Techniques; Testing; Therapeutic; Tissues; Toxic Actions; Training; Transgenic Mice; Transgenic Organisms; Viral Proteins; Woman; Work; age group; aged; associated symptom; base; biocytin; cellular imaging; chemokine; connective tissue-activating peptide; cytokine; experience; fluorescence imaging; gender difference; genetic approach; immunocytochemistry; in vivo; inhibitor/antagonist; innovation; male; men; mitochondrial dysfunction; mitochondrial membrane; mouse model; neurobehavioral; neuronal cell body; neuropathology; neuropsychiatry; neurosteroids; neurotoxic; neurotoxicity; novel; opioid abuse; opioid use; pre-clinical; programs; protective effect; receptor; relating to nervous system; research study; sex; steroid hormone; tamoxifen receptor; tissue culture; translational approach; transmission process; virotoxins

 DESCRIPTION (provided by applicant): 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. The goal of this Pathway to Independence Award (K99/R00) is to provide training to Dr. Jason Paris to discern the cellular/molecular mechanisms by which sex steroid milieu may influence opiate/HIV interactions for neuroAIDS pathology. Dr. Paris will receive advanced training in murine and human neural cell culture and imaging techniques for detection of sub lethal neuronal pathogenicity, intracellular ions, and mitochondrial membrane integrity. In the K99 training phase, Dr. Paris will work under the tutelage of Dr. Kurt Hauser (Primary Mentor) to examine the potentially-protective effects that sex steroids may exert over synergistic actions between opiates and the HIV-1 virotoxins, Tat (Aim 1a) or gp120 (Aim 1b) in murine neural co-cultures. Viable/degenerating neurons, astrocytes, and microglia will be detected via immunocytochemistry, biocytin-filled striatal medium spiny neurons (MSNs) will be assessed for sub lethal changes in morphology, and supernatants will be assessed for production of cytokines/ chemokines, reactive oxygen species, and nitrites. In murine (Aims 2a and 2b) and human (Aim 2c) neural cultures, steroid protection against opiate interactions with Tat (2a), gp120 (2b), or X4-/R5-tropic HIV (2c) will be assessed for effects on MSN morphology, ion imaging for [Ca] i and [Na] i within MSN soma and dendrites, 2+ + and mitochondrial membrane destabilization (via Rhodamine 123 fluorescence imaging). In the R00 independent phase, Dr. Paris 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). Male and female mice will be assessed for neuroAIDS-like motor, affective, and cognitive behavior. Striatal tissue will be assessed for endpoints described in Aim 1. Lastly, the influence of pharmacodynamic targets that overlap between steroids, opiates, and HIV proteins will be examined in murine neural co-cultures obtained from Tat- or gp120-transgenic mice (Aim 3c). Cultures will be pretreated with pharmacological antagonists and examined as in Aims 1 and 2. This proposal is timely, provides novel training, and will enable the development of an innovative and independent research program.

 描述（由申请人提供）：与一般人群相比，人类免疫缺陷病毒（HIV）感染者会经历与更严重的运动/情绪/认知障碍（统称为“神经艾滋病”）相关的脑部病理学，并且这些影响在阿片类药物滥用者中更为严重。此外，存在性别差异，与滥用阿片类药物的男性相比，滥用阿片类药物的女性感染艾滋病毒的风险更大；然而，一旦感染，一些女性可能会比男性经历更少的神经艾滋病症状。我们在小鼠身上重现了这些作用，并开始阐明与阿片类药物相互作用的性类固醇激素，从而为神经艾滋病提供保护。目前的提案将开始揭示性类固醇与阿片类药物和艾滋病毒相互作用的影响和机制，以利用培养的转化方法 小鼠和人类神经细胞，以及转基因全动物小鼠模型。该独立之路奖 (K99/R00) 的目标是为 Jason Paris 博士提供培训，以辨别性类固醇环境可能影响神经艾滋病病理学中阿片类药物/HIV 相互作用的细胞/分子机制。 Paris 博士将接受小鼠和人类神经细胞培养和成像技术方面的高级培训，以检测亚致死神经元致病性、细胞内离子和线粒体膜完整性。在 K99 培训阶段，Paris 博士将在 Kurt Hauser 博士（主要导师）的指导下研究性类固醇可能对阿片类药物和 HIV-1 病毒毒素、Tat (Aim 1a) 或 gp120 (Aim 1b) 之间在小鼠神经共培养物中的协同作用产生的潜在保护作用。将通过免疫细胞化学检测存活/退化神经元、星形胶质细胞和小胶质细胞，评估充满生物胞素的纹状体中型多棘神经元（MSN）的形态亚致死变化，并评估上清液细胞因子/趋化因子、活性氧和亚硝酸盐的产生。在小鼠（目标 2a 和 2b）和人类（目标 2c）神经培养物中，将评估类固醇对阿片类药物与 Tat (2a)、gp120 (2b) 或 X4-/R5-tropic HIV (2c) 相互作用的保护作用，以评估其对 MSN 形态、MSN 胞体和树突、2++ 和线粒体膜内 [Ca] i 和 [Na] i 的离子成像的影响 不稳定（通过罗丹明 123 荧光成像）。在 R00 独立阶段，Paris 博士将使用有条件表达中枢 HIV-1 Tat（目标 3a）或组成型表达中枢 gp120（目标 3b）的整体动物模型，研究中枢类固醇形成对阿片类药物/HIV 蛋白相互作用的保护作用。将评估雄性和雌性小鼠的神经艾滋病样运动、情感和认知行为。将评估纹状体组织的目标 1 中描述的终点。最后，将在从 Tat 或 gp120 转基因小鼠获得的小鼠神经共培养物中检查类固醇、阿片类药物和 HIV 蛋白之间重叠的药效学目标的影响（目标 3c）。将用药理学拮抗剂对培养物进行预处理，并按照目标 1 和 2 进行检查。该提案是及时的，提供了新颖的培训，并将有助于开发创新和独立的研究计划。