4R CEMBRANOID

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The therapy of HIV/AIDS has expanded over the years to include 26 drugs. However, rapid development of drug resistantviral strains makes antiretroviral therapy for HIV infection transiently effective in some scenarios, in others the concept of stable quiescent CD4 T cell functioning as reservoirs of the latent infection represent the mayor hurdle towards the cure of the infection. The currently used regimens of Highly Active Antiretroviral Therapy (HAART) are generally expensive, relatively toxic and carry an inefficient bioavailability particularly to the brain tissue. We understand that there is a major need to identify a compound that can prevent or alleviate the damaging effects following HIV-l infection. Continued effort for the discovery of new anti-HIV drugs, especially for new small molecules with novel anti-HIV mechanisms, is still needed. We have identified a non-toxic cembranoid called 4R that readily penetrates into the brain and has both potent anti-HIV and anti-inflammatory activity. Therefore, this proposal aims to test 4R for its ability to suppress HIV-l replication in lymphocytes from HIV-infected individuals and to modulate the production of anti-inflammatory cytokines and chemokines in the infected cells. In an attempt to understand the mechanism of these activities, we will test the following hypothesis: the novel compound 4R inhibits HIV-l replication and inflammation by interacting with the alpha7 nicotinic acetylcholine receptor which results in modulation of the antiviral Wnt/beta-catenin and the anti-inflammatory STAn pathways. The following three specific aims are proposed to test this hypothesis, using PBMC from 20 HIV-negative healthy volunteers and 30 HIV-positive patients (PBMC from HIV-negative individual will be acutely infected with HIV-1SF2 (X4/RS) dual virus strain). Aim 1. To determine whether 4R can suppress HIV replication in PBMC from HIV-infected patients using HIV-l RT PCR. Aim 2. To determine whether 4R anti-HIV activity is mediated by the modulation of HIV replication-associated cytokines and chemokines using cytometric bead array (CBA). Aim 3. To determine whether 4R anti-HIV and antiinflammatory activities are mediated by the activation of the Wnt/beta-Catenin and STAn pathways, respectively using Western blot analysis. In addition, to test whether 4R activities are due to interaction with the alpha7 nicotinic acetylcholine receptor (a7nAchR) and subsequent modulation of the intracellular calcium levels, calcium imaging will be performed.