Role of DNA sensing pathways in KSHV associated cancers

项目摘要/摘要 卡波西肉瘤相关疱疹病毒是人类几种恶性肿瘤的病原体 包括卡波西肉瘤(KS)、多中心性Castleman病(MCD)和原发性渗出性淋巴瘤(PEL)。 尽管先天免疫已被证明对清除病毒的全面抗病毒反应很重要 感染并限制病毒肿瘤的生长和增殖，这正是先天免疫系统感知肿瘤病毒的方式， 例如KSHV，在分子水平上还没有被很好地理解。研究先天免疫之间的相互作用 系统，KSHV和KSHV相关癌症将帮助我们开发更好的治疗方法，不仅治疗KSHV相关 恶性肿瘤，但也拓宽了我们对其他病毒癌症的认识。在此之前，我们已经发现cGAS-刺痛 途径负责触发KSHV感染后的先天免疫反应。然而，KSHV仍然可以 在cGAS-STING信号存在的情况下建立终身感染，提示KSHV病毒对cGAS- 抑制先天免疫的刺痛途径。为了识别潜在的病毒调节因子，我们开发了一种 筛选系统，并成功鉴定了多种负调控cGAS-STING信号的病毒蛋白。 我们还验证了我们的候选者之一vIRF1，并探索了vIRF1如何阻止cGAS- 刺痛途径促进KSHV复制。我们的结果表明，cGAS-STING途径的调制是 对病毒传播和伽马疱疹病毒在人类群体中的终生持久性很重要。 因此，K99/R00提案中的研究将建立在这些发现的基础上，以扩展我们对这些问题的了解 病毒蛋白调控cGAS-STING通路及其靶向治疗KSHV的研究进展 病毒蛋白。具体来说，我将集中在：1)验证我们的筛选结果，并探索详细的机制 我们的KSHV候选病毒的cGAS-STING信号调节，2)利用多种技术，包括 SiRNA、重组病毒和CRISPR来探索我们的候选病毒在KSHV致病中的作用 体外，3)利用CRISPR技术探索我们的候选病毒在基于PEL的小鼠模型中的作用， 4)筛选针对cGAS-STING通路的药物，并在我们基于PEL的小鼠身上测试它们的有效性 模特。凭借强大的导师和详细的培训计划，我的目标是为成功地 独立研究cGAS-STING通路与KSHV癌的相互作用 开发潜在的KSHV癌症治疗方法。

PROJECT SUMMARY/ABSTRACT Kaposi's sarcoma associated herpesvirus is the etiological agent for several malignancies in the human population including Kaposi's sarcoma (KS), Multicentric Castleman's disease (MCD), and primary effusion lymphoma (PEL). Although innate immunity has been shown to be important for comprehensive antiviral responses to clear viral infection and limit viral tumor growth and proliferation, exactly how the innate immune system senses tumor viruses, such as KSHV, is not well understood at the molecular level. Investigating the interplay between the innate immune system, KSHV and KSHV-associated cancers will help us develop better therapies to not only treat KSHV related malignancies, but also broaden our knowledge to other viral cancers. Previously, we have found that cGAS-STING pathway is responsible for triggering innate immune responses upon KSHV infection. However, KSHV can still establish lifelong infection in the presence of cGAS-STING signaling, suggesting KSHV viral regulation of cGAS- STING pathway to suppress innate immunity. In order to identify potential viral regulators, we have developed a screening system and successfully identified multiple viral proteins that negatively regulate cGAS-STING signaling. We have also validated one of our candidates, vIRF1, and explored the mechanism of how vIRF1 blocks cGAS- STING pathway to facilitate KSHV replication. Our results suggest that modulation of cGAS-STING pathway is important for viral transmission and the lifelong persistence of gammaherpesviruses in the human population. Therefore, studies in this K99/R00 proposal will build upon these findings to extend our knowledge of how these viral proteins regulate cGAS-STING pathway and how to develop potential KSHV cancer therapy by targeting these viral proteins. Specifically, I will focus on: 1) Validating our screening results and exploring detailed mechanisms of cGAS-STING signaling regulation by our KSHV viral candidates, 2) Utilizing multiple technologies, including siRNA, recombinant virus, and CRISPR to explore the role of our viral candidates on KSHV pathogenesis in vitro, 3) Utilizing CRISPR technology to explore the role of our viral candidates in our PEL based mouse model, 4) Screening for drugs that target cGAS-STING pathway, and testing their efficacy in our PEL based mouse model. With strong mentorships and a detailed training plan, I aim to build a solid foundation for a successful independent research career investigating the interaction of cGAS-STING pathway and KSHV cancers and developing potential KSHV cancer treatments.