Project Summary Even in the era of combination antiretroviral therapy (cART), people living with HIV (PLWH) exhibit a broad spectrum of neurocognitive disorders collectively termed HIV-Associated Neurocognitive Disorders (HAND). Inflammation in the brain is a hallmark of HAND, yet the mechanisms that drive this inflammation in the cART era are not entirely clear. cART penetration into the brain is lower than in other tissues and can also lead to production of noninfectious particles. Under these two scenarios, viral innate sensing may be triggered in glial cells (microglia and astrocytes) to mediate persistent inflammation. We will assess mechanisms of innate sensing in glial cells. Based on our body of work, we hypothesize that the activation of the cGAS/STING innate sensing pathway is strong and lasting after detection of HIV particles or virus like particles in glial cells, leading to inflammatory cytokine production which, in turn, will result in persistent neuroinflammation. Specifically, we will determine the impact of HIV post-entry events on innate immune sensing in glial cells (aim 1) and determine the biologic consequence of HIV-mediated innate sensing on astrocytes, either directly or through cross-talk with microglia (aim 2). We will use iPSC-derived microglia and astrocytes in our studies as well as sophisticated molecular, biochemical, and imaging tools to address our overall hypotheses. Our work will define a novel pathway that may drive and/or contribute to persistent neuroinflammation in the era of cART, which can inform targeted strategies to ameliorate and/or reduce HIV-associated neuroinflammation.