Developing Transmissible Antivirals by Exploiting Gene-Expression Circuitry

描述（由申请人提供） 翻译后摘要：新兴和成熟的病毒性疾病对人类健康造成巨大损失。目前的治疗方法不太可能阻止许多病毒，特别是HIV-1的流行病传播，原因是治疗费用（即获得）过高，合规问题，病毒快速突变以及难以接触的高风险病毒“超级传播者”的影响。我们建议将治疗模式转向开发需要病原体复制的治疗性感染性假病毒（TIP）。TIPs会沿着病原体的正常传播途径传播，准确地到达那些最需要治疗的高危人群。TIP与野生型病毒包装元件协同作用，减少体内疾病进展并减少人群规模的疾病传播。我们已经证明，抗HIV TIP可以以相同的速度突变，并在进化选择下保持其与野生型病毒的寄生关系，从而克服病毒突变逃逸。由于TIP有条件地复制（即背负式），因此消除了治疗依从性和成本问题。口服小儿麻痹症疫苗（一种减毒活疫苗）是TIPs安全性的一个先例，该疫苗的传播有限，目前正用于根除小儿麻痹症运动。为了开发候选TIPs，我们将利用我们在HIV-1转录电路方面的专业知识。我们发现HIV-1利用随机基因表达来控制进入休眠状态（前病毒潜伏期）。通过靶向对病毒反馈至关重要的细胞基因（SirT 1），我们使HIV-1偏向于休眠和减少再激活。我们将利用我们的单细胞成像方法，对促进病毒潜伏期的治疗候选药物进行高通量成像筛选，从而利用这种迫使病毒进入休眠状态的创新策略。接下来，这些候选TIP将在新型微流体恒化器中进行分析，该恒化器保持稳态感染并允许病毒在体内样环境中进化。通过将这些方法与预测模型相结合，我们将开发出一种革命性的疗法，以阻止艾滋病毒/艾滋病和其他传染病的传播。 公共卫生相关性：新出现的和确定的病毒性疾病是主要的健康问题。许多病毒性疾病缺乏有效的治疗或预防性疫苗，即使可用，这些治疗也无法阻止流行病的传播，因为病毒突变逃逸和感染性超级传播者个体的存在。显然，需要新的和更有效的抗病毒策略，该提案提出了一种多管齐下的方法来确定和开发一种创新的新的抗病毒方法。

DESCRIPTION (Provided by the applicant) Abstract: Emerging and established viral diseases take an enormous toll on human health. Current treatment approaches are unlikely to halt epidemic spread of many viruses, notably HIV-1, due to prohibitive costs of treatment (i.e. access), compliance issues, rapid viral mutation, and the influence of hard-to-reach high-risk viral 'superspreaders'. We propose to shift the treatment paradigm toward developing Therapeutic Infectious Pseudoviruses (TIPs) that require the pathogen to replicate. TIPs would transmit along a pathogen's normal transmission route, reaching precisely those high-risk populations that most require therapy. TIPs co-opt wild-type virus packaging elements, decreasing disease-progression in vivo and reducing disease transmission on a population scale. We have demonstrated that an anti-HIV TIP could mutate with equal speed and under evolutionary selection to maintain its parasitic relationship with wild-type virus, thereby overcoming viral mutational escape. Since TIPs replicate conditionally (i.e. piggyback) treatment compliance and cost issues are eliminated. A precedent for the safety of TIPs exists in the oral polio vaccine (a live-attenuated vaccine) which exhibits limited spread and is being used in the polio eradication campaign. To develop candidate TIPs we will capitalize upon our expertise in HIV-1 transcriptional circuitry. We discovered that HIV-1 exploits stochastic gene-expression to control entry into a dormant state (proviral latency). By targeting a cellular gene (SirT1) essential for viral feedback, we have biased HIV-1 toward dormancy and diminished reactivation. We will exploit this innovative strategy of forcing viruses into dormancy by utilizing our single-cell imaging methods to conduct high-throughput imaging screens for therapeutic candidates that promote viral latency. Next, these candidate TIPs will be analyzed in novel microfluidic chemostats that maintain homeostatic infection and allow viral evolution in an in vivo-like setting. By integrating these approaches with predictive models, we will develop a revolutionary therapy to halt the spread of HIV/AIDS and other infectious diseases. Public Health Relevance: Emerging and established viral diseases are major health concerns. Many viral diseases lack effective treatments or preventative vaccines and even when available these treatments are unable to halt epidemic spread due to viral mutational escape and the presence of infectious superspreader individuals. Clearly, new and more effective antiviral strategies are needed and this proposal presents a multi-pronged approach to identify and develop an innovative new antiviral approach.