Opiate abuse and sex steroids influence neuroAIDS pathology

携带人类免疫缺陷病毒(HIV)的人经历了与更大的 运动/情绪/认知障碍(统称为“神经艾滋病”)比一般人群和这些 阿片类药物滥用者的影响加剧。此外，滥用鸦片类药物的女性存在性别差异。 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更少的风险 神经艾滋病的症状比男性一旦发生感染。我们在小鼠身上重述了这些效应，并 开始阐明与鸦片类药物相互作用的性类固醇激素，以保护神经艾滋病。这个 本提案将开始揭示性激素与阿片类药物和 利用培养的小鼠和人类神经细胞以及转基因技术的翻译方法 全动物小鼠模型。这个独立之路奖(K99/R00)将开始辨别 性激素环境可能影响阿片类药物/HIV相互作用的细胞/分子机制 神经艾滋病病理学。在这笔赠款的R00独立阶段，我们将检查 体内阿片剂/HIV蛋白相互作用中中枢类固醇形成的全动物模型 有条件地表达中央HIV-1Tat(目标3a)或结构性表达中央gp120(目标3b)。神经类固醇 在雄性和雌性小鼠身上，药物将抑制或促进形成，受试者将被 评估类似神经艾滋病的运动、情感和认知行为。纹状体组织将被评估为 通过免疫细胞化学和纹状体培养液存活/退化的神经元、星形胶质细胞和小胶质细胞 刺状神经元(MSN)将被评估其形态的亚致死性变化。的影响 类固醇、阿片类药物、嗜X4/R5的HIV和HIV蛋白(TAT或 Gp120)将在获得的小鼠和分化的胎儿神经前体细胞的共同培养中进行检测。 单元格(目标3c)。培养物将用NMDA受体的药理拮抗剂mGluR1， 雌激素受体(阿尔法和贝塔)和单位阿片受体。与先前的K99项目(目标1-2)一样，文化 将评估MSN形态、MSN胞体内的细胞内离子浓度([钙]i和[Na+]i) 以及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、活性氧物种和亚硝酸盐的产生。这项建议使 开发创新和独立的研究计划。

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.