PROJECT SUMMARY The main goal of the COBRE Center is to enable and strengthen research in the area of targeted therapeutics at the University of South Carolina (USC). This is attained by offering pilot research grants to junior and more senior faculty, by supporting junior faculty through funds and mentorship, and by supporting three research cores (Functional Genomics Core, Drug Design and Synthesis Core, and Microscopy and Flow Cytometry Core). The Center for Targeted Therapeutics (CTT) brings together junior and senior faculty with a common interest in the discovery of new molecular targets and prototype drugs for the treatment of common and serious diseases. The current project is focused on finding therapeutics for the treatment of HIV-associated neurocognitive disorder (HAND) and HAND with Substance Use Disorder comorbidities (SUD). Further, the project addresses urgent public needs addressing polysubstance use of People Living with HIV. According to the World Health Organization, more than 36 million people worldwide live with HIV; of those, more than 1.2 million are in the USA. Individuals living with HIV are more likely to suffer from substance use disorder, developing a clinical condition known as HIV-Associated Neurocognitive Disorder. Drugs of abuse, specifically cocaine, fentanyl, and fentanyl-contaminated cocaine, augment HAND's severity by synergistic neurotoxicity with HIV proteins and reemergent expression of latent HIV provirus. In conjunction with SUD, HIV- associated cognitive deficiencies decrease engagement in HIV care, which fuels a downward spiral of health status. Black/African American people account for a higher proportion of new HIV diagnoses and people with HIV, compared to other races and ethnicities, and therefore suffer from HAND and SUD. Despite intensive research, there is no approved therapy for HAND, particularly for the combined neurological effects of HIV and drugs of abuse. Previously through AI-based approaches, we determined several compounds, including FDA-approved drugs, as a candidate for the repurposing for treating HAND and HAND with SUD comorbidities. However, the complexity of the disease required new models and advanced molecular technologies to determine the mechanisms of the compounds' activities. Aberrant microglial activity is an emerging target for treating neurodegenerative diseases and drug addiction, with increasing appreciation for the management of HIV-associated pathologies. Combination therapies have had tremendous success in managing HIV-1 and converting it from a death sentence to a manageable chronic disease. Similarly, combination therapies are a promising way to manage microglia status and HIV-1 expression for the treatment of HAND worsened by SUD. Understanding the effects of microglia- specific drugs required the assessment of the effects on the tissue and organism level since microglia, specifically HIV-infected microglia involved in complex interaction with brain cells: neurons, astrocytes, oligodendrocytes, and non-infected microglia, through cell-cell contacts, secreted proinflammatory factors and extracellular vesicles. Recently, humanized animal models and iPS-differentiated cerebral organoids have been suggested as promising models of HAND and SUD. The models are complementary since they allow us to determine the unique aspects of HAND: the interaction with human neurons and astrocytes in the brain organoids and the neurocognitive outcome with the humanized animals. A multidimensional collaborative approach, which involves analysis of animal behavior, neuronal structure and morphology, local and distant effects of HIV toxicity, and interactions with drugs of abuse, is necessary to understand the complexity of the disease and to develop new management strategies. The current project proposes a collaboration between groups of Dr. Michael Shtutman, director of COBRE CTT functional genomics core and expert in transcriptomics approaches and AI-based drug repurposing, Dr. Rosemarie Booze, the expert in neuroinflammation, SUD, HAND, and HAND animal models (USC), and Dr. Shilpa Buch (U. of Nebraska) the expert in HAND, SUD and cellular interactions. In the project, we will focus on the molecular mechanism of HAND and SUD, emphasizing polysubstances (cocaine and fentanyl) use disorder comorbidity of HAND and testing of AI-identified compounds. Dr. Booze's group will focus on behavioral aspects and synaptodendritic effects, Dr. Buch's group will assess the intracellular interactions and NLRP3- inflammasome pathways, and Dr. Shtutman's group will focus on integrative OMICS analysis and compounds activities on iPS differentiated brain organoids The collaborative project will help to establish a foundation for repurposing small molecule candidates for HAND/SUD treatment and to accelerate their transition to clinical studies.

PROJECT NARRATIVE Public Health Relevance: HIV Associated Neurocognitive Disorder (HAND) is one of the most common and clinically important complications of patients living with HIV infection, particularly in an aging population, and unfortunately also known to be worsened by recreational drugs. We have exploited artificial intelligence-based biomedical literature mining to find new connections and test new treatment options by “repurposing” already approved drugs for new indications. In this application, we propose to test Senicapoc, the drug developed to treat sickle-cell anemia, to alleviate or reverse HAND with substance or polysubstance use disorder comorbidities.