Abstract: Drugs of abuse are a significant comorbidity among people living with HIV. Methamphetamine (Meth), in particular, is a potent psychostimulant frequently abused in the HIV/AIDS population. Both HIV and Meth are risk factors for cognitive decline even in the era of combination antiretroviral therapy (cART). The mechanism(s) that drive and/or contribute to this cognitive decline, collectively known as HIV-Associated Neurocognitive Impairment (HAND), are not entirely clear nor is the impact of Meth on HIV reservoir. Meth itself enhances HIV replication. We will use two innovative humanized animal models to address the interface between glial cells, Meth and HIV reservoir. huAstro/HuPBMC mice, generated by engraftment of IPSC-astrocytes into NSG mice, can uniquely address the role of astrocytes as a reservoir for HIV and in HIV egress out of the brain to peripheral organs (Aim 1) and define the effect of Meth with or without HIV on prototypical functions of astrocyte and brain homeostasis (Aim 2). We focus on astrocytes because they constitute a significant resident brain cell population and perform vital functions to maintain brain homeostasis. The HuCD34/NPC model (CD34 humanized mice engrafted with neuronal progenitor cells (NPCs) will be used to assess the role of Meth on HIV evolution over time in the CNS and peripheral organs (Aim 3). Combining these two models with the resources of the Translational Methamphetamine AIDS Research Center (TMARC) and the NIDA center for genetic studies at Rutgers and cell repository (RUDCR) to reprogram lymphocytes from Meth/HIV donors to generate IPSC then NPC and/or IPSC-astrocytes as targeted for in vitro and in vivo studies provides a powerful tool to address our central hypothesis that Meth mediates a greater HIV reservoir in astrocytes and egress into peripheral organs (Aim 1), dysregulate astrocytes to disrupt brain homeostasis (Aim 2), and promote s greater extent of viral evolution within the CNS (Aim 3). Together, these studies are responsive to NIDA HIV research high priority areas and will advance our knowledge regarding the role of drugs of abuse on HIV reservoir, evolution, and neuropathogenesis to inform better strategies to uniquely address persistent HIV among the HIV positive drug abusing population.