Elucidating the mechanisms of intrinsic stem cell resistance to virus infection

阐明内在干细胞抵抗病毒感染的机制

• 批准号: 10002173
• 负责人: Xianfang Wu
• 金额: $ 9.31万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-02 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10002173
• 关键词: Advisory Committees; Affect; Antiviral resistance; Automobile Driving; Award; Basic Science; Binding; Bioinformatics; Biological Assay; Cell Differentiation process; Cells; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Complement; Counseling; Data; Data Set; Development; Development Plans; Epigenetic Process; Ethics; Evolution; Gene Expression; Genes; Genetic Transcription; Goals; Grant; Growth; Heart; Hematopoietic stem cells; Human; IRF1 gene; Immunology; Immunoprecipitation; In Vitro; Infection; Interferons; Knock-out; Laboratories; Maintenance; Malignant Neoplasms; Mediating; Mentors; Methodology; Methods; Mutagenesis; Natural regeneration; New York; Organ; Organism; Pathway interactions; Phase; Physiological; Post-Translational Protein Processing; Process; Regulation; Research; Research Personnel; Resistance; Resources; Rice; Signal Transduction; Techniques; Testing; Time; Tissues; Training; Transposase; Universities; Viral; Virus; Virus Diseases; Work; Writing; career development; cell type; combat; differential expression; epigenome; experience; experimental study; human embryonic stem cell; humanized mouse; in vivo; innovation; knockout gene; loss of function; mouse model; new technology; pathogen; permissiveness; promoter; response; skills; stem; stem cell biology; stem cell differentiation; stem cells; tissue repair; tissue stem cells; transcription factor; transcriptome; virology; virus host interaction

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.

项目概要 该独立之路提案的主要研究目标是理解和描述 干细胞中干扰素刺激基因（ISG）内在表达的机制。这 维持健康的干细胞对于生物体内的组织修复至关重要。与临终不同 然而，分化细胞不会产生同样强大的干扰素 (IFN) 反应来对抗 感染。干细胞有效阻止病毒感染的机制仍知之甚少。我们的 最近的发现表明，干细胞具有高基础水平的细胞类型特异性 ISG 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，候选人将接受以下方面的培训： 表观基因组技术和生物信息学可全局定义与本质相关的染色质可及性 在干细胞中表达 ISG。由于转录因子 (TF) 结合通常与这些可访问的 染色质结构域，对这种协调的分析将使他能够识别驱动 ISG 表达的 TF 干细胞。此外，候选人还将接受CRISPR基因敲除筛选培训，并进一步 通过这些功能丧失的补充实验来定义候选 TF。最后，候选人将是 指导新识别的 TF 以及 IRF1（一种专门的 TF）的详细机制表征 调节造血干细胞中内在的 ISG 表达。候选人的其他组成部分 全面的职业发展计划包括道德和资助写作方面的课程和研讨会。培训内容 该阶段将在洛克菲勒大学 (RU) 的 Charles Rice 博士的实验室进行，该大学是 世界领先的病毒学和免疫学研究实验室。除了大量的资源和 该实验室的基础科学专业知识，候选人还将受益于 RU 充满活力的研究 社区。职业发展的一个重要组成部分将是经验丰富的顾问的密切建议 咨询委员会，由 Charles Rice 博士、C. David Allis 博士和 Robert Roeder 博士（均为 RU）组成，以及 大卫·利维博士（纽约大学）。 K99中获得的创新技能和全面的数据集 训练阶段将为详细理解调节内在的转录网络奠定基础 ISG 表达，更重要的是干细胞的抗病毒耐药性。这将是年轻人成功的关键 这个竞争激烈的研究领域的独立研究者。总而言之，培训将满足 候选人的短期目标是在他的投资组合中添加新技术、技能和经验，以及他的长期目标 学期目标，成为一名独立调查员，研究重点是理解重要的 宿主与病毒相互作用的核心过程。这项研究将对我们产生重要影响 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。