Neuroscience
Permanent URI for this collectionhttps://hdl.handle.net/20.500.12503/21717
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Browsing Neuroscience by Author "Ghorpade, Anuja PhD"
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Item Alcohol and HIV-1 differentially regulate Toll Like Receptor (TLRs) expression and signaling in Primary Human Astrocytes(2016-03-23) Ghorpade, Anuja PhD; Pandey, RichaAbout 69% of human immunodeficiency virus-1 (HIV-1)-positive individuals exhibit some form of HIV-associated neurocognitive disorders (HAND). Several studies have reported that HIV-1 virions, viral proteins and alcohol, individually have direct or indirect effects on HAND pathophysiology. Recently, we showed that alcohol activates astrocytes and regulates inflammation via cPLA2 in HAND. Toll-like receptors (TLRs) are a family of innate immune system receptors that respond to pathogen-derived and tissue damage-related ligands. TLR signaling in immune cells, astrocytes, microglia and neurons may play roles in the pathogenesis of multiple diseases. TLRs are a “missing” link in alcohol-mediated astrocytic response in context of HAND since TLR stimulation by alcohol in glial cells induces secretion of pro-inflammatory molecules. Thus, we explored the role of TLRs in alcohol-induced inflammation and cytotoxicity in primary human astrocytes with HAND. TLRs signaling gene array was performed to screen altered profiles for all 10 TLR family members and 74 downstream signaling molecules. Ingenuity pathway analysis (IPA) was performed to identify potential signaling nodes. Data suggested that HIV-1 and/or EtOH led to differential TLRs expression in astrocytes. We confirmed all 10 TLRs by real-time PCR in four independent astrocyte donors. Alcohol alone and with HIV-1, significantly upregulated TLR1, 2, 3, 4, 5 and 9 as compared to controls and HIV-1 alone. We propose that TLRs regulation plays an important role in astrocytes inflammation upon HIV-1 and EtOH exposure.Item Astrocyte AEG-1 regulates ER stress responses in the context of HAND(2016-03-23) Ghorpade, Anuja PhD; Nooka, ShruthiEndoplasmic reticulum (ER) stress has recently been linked to neurological disorders, including HIV-associated neurocognitive disorders (HAND). We recently showed astrocyte elevated gene (AEG)-1, a multifunctional oncogene regulating astrocyte migration, proliferation and neuroinflammation. AEG-1 upregulation in Huntington’s disease model suggests its role in ER stress responses in aging and HAND. However, its involvement in ER stress responses during HIV-1 infection is not known. In this study, we investigated HIV-1 and anti-retroviral therapy (ART) drugs mediated ER stress i.e., unfolded protein response (UPR) pathway activation, and astrocyte AEG-1 expression, intracellular localization during ER stress. RT-PCR and western blot analysis revealed that HIV-1, IL-1β and ART drugs activated UPR pathway and autophagy in astrocytes. Moreover, astrocytes exposed to ER stress compounds upregulated AEG-1 expression. Confocal analysis and mPTP assay showed AEG-1 colocalization with calnexin and mitochondrial damage with ER stress. In addition, AEG-1 overexpression upregulated ER stress markers such as BiP, PERK, and CHOP that were further enhanced by IL-1β treatment. Immunocytochemical studies also showed increased autophagy markers i.e., LC3 and P62 in AEG-1 overexpressing astrocytes. In summary, our study highlights that HIV-infection and ART drugs induce ER stress in astrocytes that is further exacerbated by AEG-1. Therefore, elucidation of AEG-1 regulated UPR pathway could assist in targeting astrocyte-induced ER stress responses in HAND.