NEUROPROTECTIVES FOR HIV-ASSOCIATED NEUROCOGNITIVE DISORDERS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 人类免疫缺陷病毒（HIV）相关的神经认知障碍（HAND）是年轻人痴呆的主要原因，其特征在于通过与氧化应激、线粒体损伤和细胞凋亡相关的级联事件增加神经元死亡。本研究的目的是研究雌激素和氟西汀在HAND期间的神经保护作用。目前还没有研究调查在HAND期间神经元蛋白质组的变化，这可能会导致更好地了解雌激素和氟西汀的作用机制。我们建议使用体外模型的神经毒性模拟手挑战细胞培养与脑脊液（CSF）样本来自西班牙裔-拉丁裔纵向妇女队列库。我们假设雌激素和氟西汀将通过增强细胞存活机制（如还原/氧化途径和线粒体完整性）介导神经保护作用。我们的第一个目标是在波多黎各大学医学院建立一个体外神经毒性模型。这将通过转移约翰霍普金斯大学医学院已经设置的技术来实现。我们将验证我们队列中诊断为HAND的女性CSF的神经毒性潜力。这将通过使用神经毒性试验测量神经元死亡来实现。然后，我们将测试雌激素和氟西汀的神经保护潜力，通过预处理的神经元细胞时，挑战与CSF的手患者。这也将通过在用神经保护剂预处理后使用神经毒性测定来测量神经元死亡来实现。来自非痴呆HIV血清阳性患者的CSF将用作对照。最后，在试图了解雌激素和氟西汀的神经保护机制，我们将调查这两种化合物介导的神经元蛋白质组的变化，使用新的蛋白质组学技术，如二维差异凝胶内表达和串联质谱。该提案的短期目标是开发神经毒性模型，用于表征这些和其他具有神经保护潜力的化合物在西班牙裔HAND女性中的作用。长期目标是为PI在转化研究中提交的国家卫生研究院（NIH）K奖提案生成初步数据。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Human Immunodeficiency Virus (HIV)-associated Neurocognitive Disorders (HAND) is the mayor cause of dementia in young adults and is characterized by increased neuronal death via a cascade of events related to oxidative stress, mitochondrial damage, and apoptosis. The objective of this proposal is to study the neuroprotective potential of estrogen and fluoxetine during HAND. No studies exist investigating the changes in the neuronal proteome during HAND that may lead to a better understanding of the mechanisms of action of estrogen and fluoxetine. We propose to use an in-vitro model of neurotoxicity simulating HAND by challenging cell cultures with cerebrospinal fluid (CSF) samples from the Hispanic-Latino Longitudinal Women Cohort repository. We hypothesize that estrogen and fluoxetine will mediate neuroprotection by enhancing mechanisms of cell survival, such as reduction/oxidation pathways and mitochondrial integrity. Our first objective is to establish an in-vitro model of neurotoxicity at University of Puerto Rico Medical Science Campus. This will be accomplished by transferring techniques already set in place at Johns Hopkins University School of Medicine. We will validate the neurotoxic potential of CSF from women in our cohort diagnosed with HAND. This will be accomplished by measuring neuronal death using neurotoxic assays. We will then test the neuroprotective potential of estrogens and fluoxetine by pretreatment in neuronal cells when challenged with CSF of HAND patients. This will also be accomplished by measuring neuronal death using neurotoxic assays after pretreatment with neuroprotectors. CSF from non-demented HIV-seropositive patients will be used as controls. Finally, in an attempt to understand the mechanisms of neuroprotection of estrogens and fluoxetine, we will investigate changes in the neuronal proteome mediated by these two compounds using novel proteomics techniques such as two-dimensional differential in-gel expression and tandem mass spectrometry. The short-term goal of this proposal is to develop neurotoxic models for the characterization of these and other compounds with potential for neuroprotection in Hispanic women with HAND. The long-term goal is to generate preliminary data for a National Institute of Health (NIH) K award proposal submission by the PI in translational research.