Methamphetamine Activation of Inflammasome and Altered immunity in HIV (MAIA) Study - PROJECT SUMMARY/ ABSTRACT Despite over four decades of research, we still do not have an HIV cure. While antiretroviral therapy (ART) is able to suppress virus to undetectable levels, it is not a cure; virus rapidly rebounds from latently-infected cells (“the HIV reservoir”) within weeks after ART interruption. Thus, a major goal is to eradicate the HIV reservoir in order to achieve clinical remission. However, high-risk individuals, such as those who use methamphetamine (MA), are most likely to benefit from HIV cure strategies and yet often have high rates of suboptimal ART adherence and poorer clinical outcomes. Recent preliminary work from our group showed for the first time that MA is associated with increased inflammation (e.g., via the NOD-like receptor family pyrin domain containing 3, NRLP3, inflammasome activation pathway) and residual HIV transcription. The proposed work now aims to identify how MA-induced inflammasome activation leads to immune dysfunction in PWH, even during ART suppression. Leveraging rare existing longitudinal blood samples from PWH on ART with and without MA (MA+/MA-) use from the UCSF EMRLHD (Effect of Methamphetamine on Residual Latent HIV Disease) study, as well as prospectively collected blood and cerebrospinal fluid (CSF), we will test whether MA promotes immune dysfunction in PWH on ART via a multi-step process: (1) alteration of metabolism, leading to (2) epigenetic changes, which then promotes (3) enhanced inflammasome-mediated signaling and (4) antiviral interferon (IFN) signaling, resulting in (5) altered immune dysfunction. Using advanced single cell methods (e.g., single cell sequencing of RNA and chromatin accessibility, high-dimensional flow cytometry), methods which are increasingly being used to prioritize targeted cell-based therapeutics, we will (a) directly test and validate that MA exposure leads to inflammasome activation and inflammation (Aim 1), and then subsequently promotes immune dysfunction in T cells (Aim 2) and B cells (Aim 3). In Aim 1 we will determine whether MA alters induces inflammasome activation and inflammation secondary to MA-induced metabolic (e.g., metabolomics) and epigenetic (e.g., scRNA/ATAC-seq) alterations, perform in vitro validation of our findings. In Aim 2, we will then determine whether MA confers antigen-specific T cell dysfunction and HIV reservoir activation, performing in vitro validation to determine the magnitude of MA-induced HIV gag-specific responses, cytokine production, histone modification, immune dysfucntion (PD-1, TOX, IRF4), and viral reservoir reactivation (LARA assay). In Aim 3, we will determine whether MA leads to MA-induced post-translational antibody modification (e.g., glycosylation, sialylation) and Fc dysfunction (e.g., ADCC, ADCP, ADNP), performing in vivo validation comparing the magnitude of antibody neutralization from pre- and post-vaccination blood and CSF samples. The proposed work will generate critical data informing how MA-induced activation of the inflammasome promotes altered immunity in PWH on ART, and provide a framework by which to study additional therapeutic targets for reducing inflammation and morbidity (and potentially, inducing HIV remission) in all PWH.
