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

基本信息

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
项目类别：
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

项目摘要

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（由适用提供）：患有人类免疫缺陷病毒（HIV）的个体经历了与一般人群相比，与更多的运动/情绪/认知障碍相关的脑病理学（集体称为“神经辅助”），并且这些作用在操作施虐者之间加剧了。此外，与优化虐待男性相比，优化虐待妇女的虐待妇女存在性别差异。然而，有些女性可能会比一旦感染发生的男性症状较小。我们在小鼠中概括了这些作用，并已开始阐明与之相互作用的性别立体骑兵扩展以赋予神经助剂的保护。本提案将开始揭示性别立体相互作用与优化和艾滋病毒的效果和机制，该方法利用了培养的方法鼠和人类神经元细胞，以及转基因全动物鼠模型。该途径获得独立奖的目标（K99/R00）是向杰森·巴黎博士提供培训，以辨别性类固醇基环境可能影响神经辅助病理学的手术/HIV相互作用的细胞/分子机制。巴黎博士将接受有关鼠和人类神经元细胞培养和成像技术的高级培训，以检测杀致命的神经元致病性，细胞内离子和线粒体膜完整性。在K99培训阶段，巴黎博士将在Kurt Hauser博士（主要导师）的指导下进行工作，以检查性别立体定子可能对Murine Neuronal co-coultire的性行为和HIV-1 Virotoxins，TAT（AIM 1A）或GP120（AIM 1A）（AIM 1A）或GP120（AIM 1B）之间的协同作用所产生的潜在保护作用。将通过免疫细胞化学，生物环蛋白填充的纹状体培养基神经元（MSN）检测到可行/退化的神经元，星形胶质细胞和小胶质细胞，将评估形态学的杀伤性变化，并评估上清液的产生，以用于产生细胞因子/趋化因子，反应性氧气，反应性氧化物种，NITIR and NITRIT和NITITIR。 In murine (Aims 2a and 2b) and human (Aim 2c) neuronal cultures, steroid protection against active 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不稳定（通过Rhodamine 123荧光成像）。在R00独立阶段，巴黎博士将使用有条件表达表达的中央HIV-1 TAT（AIM 3A）或构造表达的中央gp120（AIM 3B）的全动物模型，研究中央立体构成对体内开放/HIV蛋白相互作用的受保护作用。将评估男性和雌性小鼠的神经辅助运动，情感和认知行为。将评估AIM 1中描述的端点的纹状体组织。最后，将在鼠神经元共培养中检查类固醇，操作和HIV蛋白之间重叠的药效靶标的影响，该靶标在鼠神经元共培养中检查了从TAT或GP120-GP120-TERANSGENIC小鼠获得的（AIM 3C）。培养物将与药理学拮抗剂进行预处理，并像目标1和2所示一样进行检查。该建议是及时的，提供了新颖的培训，并能够开发创新和独立的研究计划。