Elucidating the mechanisms of intrinsic stem cell resistance to virus infection

项目摘要 这条通往独立的道路提案的主要研究目标是理解和描述 干扰素刺激基因(ISGs)在干细胞中内在表达的机制。这个 维持健康的干细胞对于生物体内的组织修复是必不可少的。与终端不同 然而，分化后的干细胞不会产生同样强大的干扰素(干扰素)反应来对抗 感染。干细胞有效阻止病毒感染的机制仍然知之甚少。我们的 最近的发现表明，干细胞具有高基础水平的特定细胞类型的ISGs亚群， 提供针对多种病毒的有效保护。这种内在ISG表达的机制 仍然难以捉摸。为此，在本次K99奖项的指导阶段，候选人将接受 表观基因组技术和生物信息学全球定义染色质可及性 在干细胞中表达ISGs。因为转录因子(TF)结合通常与这些可访问的 染色质结构域，对这种配位的分析将使他能够识别驱动ISG表达的转录因子 干细胞。此外，候选人将接受CRISPR基因敲除筛查的培训，并进一步 用这些功能丧失的互补实验来定义候选的TF。最后，候选人将是 指导新发现的TF和IRF1的详细机械特性，这是一种专门的TF 调节造血干细胞内固有的ISG表达。候选人的其他组件 全面的职业发展计划是关于道德和拨款撰写的课程和研讨会。培训 这一阶段将在洛克菲勒大学查尔斯·赖斯博士的实验室进行，洛克菲勒大学是 世界领先的病毒学和免疫学研究实验室。除了重要的资源和 在这个实验室的基础科学专业知识，候选人也将受益于RU充满活力的研究 社区。职业发展的一个关键组成部分将是经验丰富的 咨询委员会，由查尔斯·赖斯博士、C.David Allis博士和Robert Roeder博士(均为RU)组成，以及 戴维·利维博士(纽约大学)。在K99中获得的创新技能和全面的数据集 培训阶段将为详细了解调节内在基因的转录网络奠定基础 ISG的表达，以及更重要的是，干细胞中的抗病毒耐药性。这将是年轻时取得成功的关键 在这一竞争激烈的研究领域中的独立调查者。总的来说，培训将满足两个方面的要求 候选人的短期目标是将新技术、技能和经验添加到他的投资组合中，以及他的长期目标 学期目标，成为一名独立的研究人员，专注于了解重要的 处于主机与病毒交互的核心的进程。这项研究将对我们的 了解干细胞生物学，发育和癌症的原始方面，以及干细胞的进化 脊椎动物病原体防御。

Project Summary The primary research goal of this Pathway to Independence proposal is to understand and characterize the mechanisms underlying intrinsic expression of interferon stimulated genes (ISGs) in stem cells. The maintenance of healthy stem cells is essential for tissue repair within an organism. Unlike terminally differentiated cells, however, stem cells do not produce the same robust interferon (IFN) response to combat infection. The mechanisms by which stem cells potently block viral infection are still poorly understood. Our recent discoveries demonstrate that stem cells have high basal levels of cell type-specific subsets of ISGs that confer potent protection against a number of viruses. The mechanisms underlying this intrinsic ISG expression remain elusive. To this end, in the mentored phase of this K99 award, the candidate will be trained in epigenome techniques and bioinformatics to globally define chromatin accessibility associated with intrinsically expressed ISGs in stem cells. As transcription factors (TFs) binding is often associated with these accessible chromatin domains, the analysis of such coordination would allow him to identify TFs driving ISG expression in stem cells. In addition, the candidate will receive training on CRISPR gene knockout screens, and further define candidate TFs with these loss-of-function complementary experiments. Finally, the candidate will be guided on detailed mechanistic characterization of newly identified TFs as well as IRF1, a TF specifically regulates intrinsic ISG expression in hematopoietic stem cells. Additional components of the candidate's comprehensive career development plan are courses and seminars in ethics and grant writing. The training phase will be carried out in the laboratory of Dr. Charles Rice at The Rockefeller University (RU), one of the world's leading laboratories in virology and immunology research. In addition to the significant resources and basic science expertise in this laboratory, the candidate will also benefit from RU's vibrant research community. A critical component of career development will be the close counsel of a highly experienced Advisory Committee, composed of Dr. Charles Rice, Dr. C. David Allis, and Dr. Robert Roeder (all RU), and Dr. David Levy (New York University). The innovative skills and comprehensive datasets obtained in the K99 training phase will set the stage for detailed understanding of the transcriptional networks that regulate intrinsic ISG expression, and more importantly, antiviral resistance in stem cells. This will be key to succeed as a young independent investigator in this highly competitive field of research. In all, the training will fulfill both the candidate's short-term goals of adding new technologies, skills, and experience to his portfolio, and his long- term goals, to become an independent investigator with a research focus on understanding important processes at the heart of host-virus interactions. This study will have important implications for our understanding of stem cell biology, primordial aspects in development and cancer, and the evolution of vertebrate pathogen defense.