HSF1作为新型HIV潜伏感染激活剂药物靶点的研究

The persistence of HIV-1 latent reservoirs is a major barrier ofcombined antiretroviral therapy (cART) to achieve a complete cure for HIV. The strategy "shock and kill" using HIV latency reversing agents (LRAs) can achieve the functional cure of HIV by reactivating the HIV proviruses integrated in the host genome and eliminating latent viral reservoirs. In the previous study, we have demonstrated that heat shock factor 1 (HSF1) can bind to the HIV 5’-LTR and positively participate in latent HIV reactivation. In this project, we intend to further reveal the molecular mechanisms of HSF1 in regulating the reactivation of latent HIV, and explore the potential of HSF1 as a novel drug target of HIV latent reversing agents. Our major research contents are: 1) to investigate the role of HSF1 in latency reactivation by CRISPR/cas9; 2) to find out compounds which specifically target HSF1 by HEK-293T-HSF1Luciferase cells and SPR chip screening, then evaluating their abilities to reactivate latent HIV and clarifying the feasibility of HSF1 as a new LRAs drug target; 3) to synthesize a novel latent HIV reversing agent by structural optimization of a positive compound resveratrol; 4) to investigate the mechanism of the highly reactive compounds, expound its key steps and sites in activation of HSF1. This project aims to validate HSF1 as a novel drug target, and obtain new LRAs which targeting HSF1 for eradication the resevoir of HIV latency and achieving a final functional cure of HIV/AIDS.

HIV潜伏病毒储存库的存在是导致抗逆转录药物无法治愈艾滋病的主要原因。采用“激活并杀灭”的策略，通过活化HIV前病毒，清除潜伏病毒库，可望实现功能性治愈。本课题组此前发现HSF1可结合HIV 5’-LTR，参与潜伏活化过程。本项目拟深入揭示HSF1调控HIV潜伏感染活化的分子机制，并探讨其作为新型潜伏感染激活剂药物靶点的可能性。主要研究内容包括：1）采用基因敲除等手段明确HSF1在潜伏感染活化中的作用；2）采用高通量筛选，寻找特异性作用于HSF1的化合物，评价其激活潜伏HIV的能力，验证HSF1作为药物新靶点的可行性；3）对白藜芦醇进行结构优化，得到特异性靶向HSF1的高活性型潜伏感染激活剂；4）阐明高活性化合物作用于HSF1的环节及关键位点。本项目旨在确证HSF1为药物新靶点，获得以HSF1为靶点的原创性HIV潜伏感染激活剂，期望其清除HIV潜伏感染库，实现HIV功能性治愈。

艾滋病领域普遍认为使用“激活-杀灭”（Shock and Kill）策略能够实现艾滋病的功能性治愈。即使用潜伏感染激活剂（Latency reversing agents，LRAs）激活潜伏感染细胞中的HIV整合基因，使之重新转录和翻译，之后再联合cART阻止病毒的二次感染；同时活化的HIV颗粒诱导细胞病变效应和天然免疫系统对细胞特异性杀伤，实现HIV的功能性治愈。.我们此前研究发现HSF1可以通过发生磷酸化形成三聚体后入核，同时募集p300来自身乙酰化，进一步的与HIV LTR区的HSE结合，诱导转录起始。同时HSF1还募集p-TEFb产生转录延伸的作用。.本项目旨在确证HSF1作为新型HIV潜伏感染激活剂的药物作用靶点，开发出以HSF1为靶点的原创性高效安全的新型HIV潜伏感染激活剂。该项目执行期间我们发表署该项目批准号（81773787）文章10篇，其中SCI论文10篇。