Elimination of the persistent reservoir of resting latently HIV-infected cells is the greatest challenge to HIV eradication. Estimates indicate that complete clearance of HIV from the body would require >70 years of continuous fully suppressive antiretroviral treatment (ART). However, induction of HIV from its dormant state could accelerate the decline of the latent reservoir shortening the time to eradication. Initial efforts to induce HIV focused on latency reversing agents (LRAs) that stimulate HIV transcription by activating T cells. However, in clinical trials these agents failed to reduce the HIV reservoir and were toxic. Recent efforts have focused on polyclonal T cell stimulants at doses that induce minimal-to no-side effects in patients (e.g. PKC agonists) or agents that use alternative pathways to induce HIV transcription without T cell activation (e.g. HDAC inhibitors). However, while transient increases in HIV in peripheral blood (PB) have been detected in clinical trials, no LRA has reduced the latent HIV reservoir in vivo. Furthermore, previous attempts to induce HIV from patients and animal models focused on PB. Their systemic effect in tissues has been poorly described. Our long-term goal is to accelerate HIV eradication by identifying LRAs that are easy to administer, safe, have little-to-no immune stimulatory activity, induce HIV in PB and tissues, and effectively reduce the latent HIV reservoir. Recently, it was shown that induction of the non-canonical NF-B (ncNF-B) signaling pathway by AZD5582, a peptide mimetic with second mitochondrial activator of caspases (SMAC)-like activity, reverses HIV latency in vitro and ex vivo. Based on these data, we evaluated AZD5582 in vivo in BLT humanized mice and observed robust target cell engagement and HIV induction. Our preliminary data shows that AZD5582 administration to ART-suppressed BLT mice induced reproducible plasma HIV RNA. These results were confirmed in SIV-infected, ART suppressed non-human primates. Analysis of resting CD4+ T cells from AZD5582-treated BLT mice revealed robust (up to 24 fold) systemic HIV induction in virtually all tissues analyzed including lymph nodes, thymus, bone marrow, liver, and lung. Importantly, we observed no overt toxicity or T cell activation. To our knowledge, this is the first demonstration of widespread HIV induction in resting cells from tissues by a LRA. Based on our preliminary data, our hypothesis is that SMAC mimetics will induce HIV expression in resting latently infected CD4+ T cells in vivo thereby reducing the latent HIV reservoir. Our objective here is to evaluate and compare the in vivo LRA activity of a panel of SMAC mimetics by analyzing their: 1) PK profile, safety and target engagement in vivo, 2) effect on HIV RNA levels in plasma and in latently-infected resting CD4+ T cells from tissues, and 3) effect on the latent HIV reservoir in vivo. These data will be used to select the best candidates for further evaluation leading to...