Investigation On The Mechanism Of Daphne Diterpene NB6 And Resveratrol Activating Latent HIV

Although highly effective antiretroviral therapy(HAART)has a good effect on the control of HIV replication and transmission,patients need to receive HAART for life to ensure the suppression of the virus,for HAART cannot completely eliminate the HIV with replication ability but in a latent state."Shock and Kill"therapy aims to reactivate the latent HIV in cells through latency reverse agents,and then combines with HAART and HIV specific immune cells to kill the cells producing HIV virus,so as to completely remove the latent reservoir of HIV.Based on of"Shock and Kill"therapy,the first step is to find a highly effective and low-toxic HIV latency activator,which can effectively activate the latent HIV without causing global activation of T cells.However,most of the reported clinical trials on HIV latency activators are dissatisfactory.Therefore,for the practical application of"Shock and Kill therapy",it’s urgent to find and develop more safe and effective HIV latency activators.By testing daphne type diterpenes isolated from the herb daphne,we screened a compound NB6 that could effectively activate the expression of latent HIV at low concentrations(10 nM).Through the study of its mechanism,we found that NB6 activated latent HIV through the PKC-NF-κB signaling pathway,further research showed that NB6 could also promote ELL2 increases by PKC pathway,and promote ELL2-SEC complex assembly and Tat to recruit more ELL2-SEC,these findings gave us a more comprehensive understanding of the mechanism of PKC agonists on reactivating latent HIV virus.Further studies showed that NB6 did not cause the activation of human CD4+T cells at 10nM and promoted latent HIV transcription of T cells from HIV patients after HAART treatment.This result confirmed that NB6 also had an efifect on the activation of latent HIV in reality.Through the research on resveratrol(RES)in this project,we found that RES could relieve the inhibition of Tat by activating AKT/FoxO1 signaling pathway,which promoted the expression of Tat protein.In addition,we found that RES destroyed the formation of SEC complex in the absence of Tat.While in the presence of Tat,RES promoted Tat to recruit more SEC components,which has a positive impact on the transcription elongation of HIV genes.Similarly,we found that inhibition of AKT significantly inhibited the activation of RES in latent HIV while inhibition of FoxO1 significantly promoted the activation of RES in latent HIV of 2D 10 cells.All these above indicated that AKT/FoxO1 played an important role in RES’s ability of antagonizing latent HIV.Overall,through screening and discovering new HIV latent activators from traditional herbs,this study provides a new scheme and a research foundation for the search and development of drugs to treat HIV.