Although combinations of anti-HIV drugs can effectively suppress virus replication, a reservoir of cells harboring latent HIV-1 precludes a cure of infection. The best described latent reservoir consists of long-lived memory CD4+ T cells that contain a transcriptionally silent but replication-competent provirus. Multiple mechanisms are involved in the establishment and maintenance of latency, especially limiting levels of the key RNA Polymerase II elongation factor termed P-TEFb. Core P-TEFb consists of Cyclin T1 as a regulatory subunit and CDK9 as the catalytic subunit. The proposed research will optimize and perform an imaging- based automated screen for compounds that induce P-TEFb in resting CD4+ T cells. Compounds that score positive in this screen will be validated in immunoblots for the ability to induce P-TEFb in resting CD4+ T cells, as well as for their ability to function as latency reversal agents in a primary CD4+ T cell model of latency. Compounds identified by this novel screen have the potential to contribute to HIV cure strategies.
We will establish and perform an imaging-based automated screen for compounds that induce P-TEFb in resting CD4+ T cells from healthy blood donors. P-TEFb consists of Cyclin T1 and CDK9 and down- regulation of this RNA Polymerase II elongation factor occurs in resting CD4+ T cells. Because induction of P-TEFb in resting CD4+ T cells is required for reactivation of latent HIV, the goal of our screen is to identify novel compounds that act as HIV latency reversal agents and therefore have potential in HIV cure strategies.
