_ This program project application comprises a team of investigators with complimentary research strengths and resources that will be harnessed for the development of novel small molecules that antagonize the action of the HIV-1 vif protein. Project 2 (Virologic support for SAR and mechanisms of inhibitor resistance) will predominantly support drug discovery/SAR efforts in Project 1 as well as identifying mutations conferring inhibitor resistance in vitro and in vivo. Specific responsibilities of Project 2 include: ? Evaluation of antiviral activity of lead compounds and their analogs in permissive and non-permissive cells. The antiviral activities of analogs emerging from the SAR exercise of Project 1 will be compared in permissive cells (vif-independent replication) and non-permissive cells (vif-dependent replication) and will be used to drive the design of analogs with more potent antiviral activity. ? Evaluation of antiviral activity in primary cells relevant to CNS reservoirs of viral replication. Macrophage are the principle cellular targets in the CNS and infected macrophage drive the neuropathogenic manifestations of viral replication. Therefore, vif antagonists will be evaluated for antiviral activity in primary macrophage in vitro. ? Identification of mechanisms governing inhibitor resistance in vitro and in vivo. HIV-1 will be passaged in non-permissive cells in increasing concentrations of prioritized Vif analogs in order to derive inhibitor resistant virus. In addition, plasma viral RNA from inhibitor-treated macaques (Project 3) will be cloned and sequenced to identify mutations conferring resistance in vivo. Impact of vif-inhibitor resistance on sensitivity to other ARV classes will be conducted with Core B. The virologic studies outlined in Project 2 will guide the SAR studies and drive the prioritization of analogs that go forward for analysis of in vivo efficacy in macaques.
Our goal is to derive potent vif antagonists through SAR analysis. SAR will require a robust assay with which to identify analogs that inhibit viral replication in a vif-dependent manner. Project 2 will assess antiviral potency of analogs emerging from the SAR exercise of Project 1 using an assay that strictly gauges vif dependent antiviral antagonism. Project 2 will also assess antiviral activity in macrophage which are the most relevant cell in the CNS. These parameters will prioritize analogs that undergo in vivo evaluation.
|Sattentau, Quentin J; Stevenson, Mario (2016) Macrophages and HIV-1: An Unhealthy Constellation. Cell Host Microbe 19:304-10|
|Rainho, Jennifer N; Martins, Mauricio A; Cunyat, Francesc et al. (2015) Nef Is Dispensable for Resistance of Simian Immunodeficiency Virus-Infected Macrophages to CD8+ T Cell Killing. J Virol 89:10625-36|
|Stevenson, Mario (2015) Role of myeloid cells in HIV-1-host interplay. J Neurovirol 21:242-8|
|Yang, Chao-Shun; Rana, Tariq M (2013) Learning the molecular mechanisms of the reprogramming factors: let's start from microRNAs. Mol Biosyst 9:10-7|
|Shen, Yang; Altman, Michael D; Ali, Akbar et al. (2013) Testing the substrate-envelope hypothesis with designed pairs of compounds. ACS Chem Biol 8:2433-41|
|Silver, Nathaniel W; King, Bracken M; Nalam, Madhavi N L et al. (2013) Efficient Computation of Small-Molecule Configurational Binding Entropy and Free Energy Changes by Ensemble Enumeration. J Chem Theory Comput 9:5098-5115|
|Nalam, Madhavi N L; Ali, Akbar; Reddy, G S Kiran Kumar et al. (2013) Substrate envelope-designed potent HIV-1 protease inhibitors to avoid drug resistance. Chem Biol 20:1116-24|
|Mohammed, Idrees; Parai, Maloy K; Jiang, Xinpeng et al. (2012) SAR and Lead Optimization of an HIV-1 Vif-APOBEC3G Axis Inhibitor. ACS Med Chem Lett 3:465-469|
|Ali, Akbar; Wang, Jinhua; Nathans, Robin S et al. (2012) Synthesis and structure-activity relationship studies of HIV-1 virion infectivity factor (Vif) inhibitors that block viral replication. ChemMedChem 7:1217-29|