RNAi for the Treatment of Viral Hepatitis

描述（由申请人提供）：这项工作的目标是推进我们关于开发基于RNAi的临床治疗所需的科学原理的原始研究。我们的计划是为安全有效的基于转录的RNA干扰（RNAi）提供一个平台，使其进入肝脏等重要器官。我们特别感兴趣的是开发一种治疗丙型肝炎病毒感染（HCV）的方法，这仍然是一个未满足的临床需求。直到最近，这一领域的研究一直受到阻碍，缺乏能力，研究复制从一个完整的病毒感染细胞培养和缺乏动物模型。因此，HCV复制子已成为研究病毒复制的主要手段。我们已经发现，一些RNAi靶标对复制子有效，但对共享相同靶序列的真正病毒感染相对惰性。我们计划根据我们的新模型和关于shRNA载体生物学的信息，重新评估HCV基因组的各个区域的RNAi敲除。此外，我们将试图明确评估靶向HCV负链的有效性，因为它在感染细胞中的产生量要低得多。我们将确定同时针对这两条链是否会提供更好的保护，并减少准物种形成的可能性。我们计划使用我们新开发的重组AAV载体的细胞培养和人源化小鼠肝脏模型，并确定RNAi作为防止HCV暴露以及在建立持续感染后消除病毒的手段的有效性。到授权期结束时，我们相信我们将能够提供关于体内有效的基于载体的RNAi所需参数的重要见解，并具有作为单一疗法和/或与其他非核酸疗法组合治疗HCV感染的稳健的临床相关手段。 公共卫生相关性：这项工作的目标是发展科学的原则，并铺平道路，开发一个安全有效的临床治疗丙型肝炎病毒感染的RNA干扰（RNAi）。RNAi是一种强大的新技术，可以关闭不需要的基因，例如病毒病原体中存在的基因。将在新的临床相关细胞培养和HCV感染的啮齿动物模型中测试新的抗HCV RNAi治疗剂消除和/或预防HCV感染的能力。

DESCRIPTION (provided by applicant): The goal of the work is to advance our original studies on developing the scientific principles required for RNAi-based clinical therapeutics. Our plan is to provide a platform for safe and effective transcription-based RNA interference (RNAi) into vital organs such as the liver. We are specifically interested in developing a therapeutic approach for Hepatitis C Virus Infection (HCV) for which there is still an unmet clinical need. Until recently, research in this area has been hampered by the lack of ability to study replication from an intact viral infection in cell culture and the paucity of animal models. As a result, HCV replicons have been the primary means to study viral replication. We have discovered that some RNAi targets are effective against replicons but relatively inert against bona-fide viral infection that shares identical target sequences. We plan to re-evaluate various regions of the HCV genome for RNAi knockdown based on our new models and information regarding shRNA-vector biology. In addition, we will attempt to definitively evaluate the effectiveness of targeting the HCV minus strand because it is produced in much lower amounts in infected cells. We will determine if targeting both strands simultaneously will provide better protection and lessen the likelihood of quasi-species formation. We plan to use cell culture and humanized mouse liver models with our newly developed recombinant AAV vectors and determine the validity of RNAi as a means to protect against HCV exposure as well as elimination of the virus after establishment of a sustained infection. By the end of the granting period, we believe we will be able to provide important insights into the parameters required for effective vector-based RNAi in vivo and have a robust clinically relevant means of treating HCV infection as a single therapy and/or in combination with other non-nucleic acid based therapeutics. PUBLIC HEALTH RELEVANCE: The goal of this work is to develop the scientific principles and pave the way to develop a safe and efficacious clinical therapy for Hepatitis C virus infection using RNA interference (RNAi). RNAi is a powerful new technology for turning off unwanted genes such as those present in a viral pathogen. The new anti-HCV RNAI therapeutics will be tested for their ability to eliminate and/or prevent HCV infection in newer clinically relevant cell culture and rodent models of HCV infection.