Targeted CRISPR/Cas Editing to Eliminate SIV CNS Reservoir in Methamphetamine-Exposed Rhesus Macaques on ART - Summary In the ongoing battle against HIV, current antiretroviral therapies (ART) fall short of providing a cure, primarily due to latent reservoirs in the body, particularly in the central nervous system (CNS). Myeloid cells, including microglia and macrophages (MM), serve as key cellular reservoirs in the CNS, perpetuating the infection by releasing toxic viral proteins and inflammatory mediators. This persistent infection is further complicated by substance use disorders (SUD), with methamphetamine (METH) use disorder (MUD) being notably prevalent. METH exacerbates the situation by disrupting the blood-brain barrier (BBB), activating microglia, and triggering the NLRP3 inflammasome, which leads to increased neuroinflammation and neuronal damage. Recent advancements in CRISPR/Cas genome editing offer a glimmer of hope. This technology has shown promise in preclinical studies for eradicating HIV provirus from various tissues, including the brain. The proposed research aims to harness this potential by using a novel AAV serotype (R2Mac) capable of penetrating the BBB and targeting MM. This serotype will deliver multiplex Sso7d-mediated enOsCas12f (SOS12f) editors to eliminate HIV/SIV provirus and the NLRP3 inflammasome. The research will be conducted using a Rhesus Macaque model of SHIV infection on ART, which closely mimics human conditions. The study will leverage the combined expertise of Dr. Ling and Dr. Hu in nonhuman primate models, METH neurotoxicity, MM inflammasome, CRISPR/Cas HIV eradication, and targeted gene therapy. The proposal includes two specific aims: first, to determine the efficiency and specificity of the R2Mac-mediated multiplex SOS12f gene therapy in eliminating SHIV provirus and inflammasome in SHIV-infected MM; and second, to evaluate the therapeutic efficacy of this approach at analytical ART interruption (ATI) using the SHIV+ART+ model, both with and without METH administration. This innovative approach, utilizing the R2Mac serotype and multiplex editing, holds the potential to significantly advance our understanding and treatment of NeuroHIV and MUD, offering new hope for those affected by these challenging conditions.
