HIV/AIDS is a lifelong disease with global impact. Despite the development of effective therapies, there is an ongoing need to identify novel drug targets for improving the treatment of HIV infection. A poorly understood area of HIV biology is the stage in the virus infection process termed uncoating, which involves disassembly of the polymeric viral capsid from the viral core after entry into the cell. In collaboration with Pfizer Global Research and Development, we have begun to characterize the mechanism of novel small molecule HIV-1 inhibitors that target the viral capsid. Preliminary results indicate that these inhibitors block HIV-1 infection at an early stage by triggering premature uncoating of the virus in the target cell. This proposal includes a comprehensive plan involving the tools of structural biology, cell biology, and virology to define the molecular target and detailed mechanism of action of such capsid-targeting HIV-1 inhibitors.
The Specific Aims are: 1. To determine the structural consequences of PF-03450074 binding to the HIV-1 capsid. 2. To determine whether PF-03450074 promotes premature HIV-1 uncoating in target cells. 3. To determine the spectrum of mutations that confers resistance to PF-03450074. 4. To determine the mechanistic basis for the effects of the cyclophilin A-CA interaction on HIV-1 sensitivity to PF-03450074. 5. To determine the molecular basis of the late-stage inhibition by PF-03450074. The proposed research will identify a novel mechanism to inhibit HIV-1 infection, will reveal the structure of the target thereby facilitating the design of lead compounds for drug development, and will reveal new insights into the biology of HIV-1 infection.
Effective treatment of HIV/AIDS requires the development of novel antiviral compounds that can complement the existing drug arsenal. This research project will define the mechanism of antiviral compounds acting on a novel HIV-1 target-the capsid. The studies proposed herein will facilitate the development of novel therapies and help elucidate the stage of HIV-1 infection termed uncoating.
|Hulme, Amy E; Kelley, Z; Foley, Deirdre et al. (2015) Complementary Assays Reveal a Low Level of CA Associated with Viral Complexes in the Nuclei of HIV-1-Infected Cells. J Virol 89:5350-61|
|Shi, Jiong; Zhou, Jing; Halambage, Upul D et al. (2015) Compensatory substitutions in the HIV-1 capsid reduce the fitness cost associated with resistance to a capsid-targeting small-molecule inhibitor. J Virol 89:208-19|
|Zhou, Jing; Price, Amanda J; Halambage, Upul D et al. (2015) HIV-1 Resistance to the Capsid-Targeting Inhibitor PF74 Results in Altered Dependence on Host Factors Required for Virus Nuclear Entry. J Virol 89:9068-79|
|Halambage, Upul D; Wong, Jason P; Melancon, Bruce J et al. (2015) Microplate-based assay for identifying small molecules that bind a specific intersubunit interface within the assembled HIV-1 capsid. Antimicrob Agents Chemother 59:5190-5|
|Price, Amanda J; Jacques, David A; McEwan, William A et al. (2014) Host cofactors and pharmacologic ligands share an essential interface in HIV-1 capsid that is lost upon disassembly. PLoS Pathog 10:e1004459|
|Lamorte, Louie; Titolo, Steve; Lemke, Christopher T et al. (2013) Discovery of novel small-molecule HIV-1 replication inhibitors that stabilize capsid complexes. Antimicrob Agents Chemother 57:4622-31|
|Yang, Haitao; Ji, Xiaoyun; Zhao, Gongpu et al. (2012) Structural insight into HIV-1 capsid recognition by rhesus TRIM5?. Proc Natl Acad Sci U S A 109:18372-7|
|Du, Shoucheng; Betts, Laurie; Yang, Ruifeng et al. (2011) Structure of the HIV-1 full-length capsid protein in a conformationally trapped unassembled state induced by small-molecule binding. J Mol Biol 406:371-86|
|Shi, Jiong; Zhou, Jing; Shah, Vaibhav B et al. (2011) Small-molecule inhibition of human immunodeficiency virus type 1 infection by virus capsid destabilization. J Virol 85:542-9|