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.
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