Molecular Programming of Immunological Memory in Human Natural Killer Cells

 描述(由申请人提供)：项目摘要最近的证据表明，自然杀伤(NK)细胞可以形成对病毒抗原的免疫记忆。我们广泛的初步数据表明，巨细胞病毒(CMV)感染与高分化、适应性NK细胞新群体的产生有关。这些NK细胞亚群显示了全基因组的DNA甲基化图谱，反映了效应性CD8T细胞的DNA甲基化图谱。在目前的提案中，申请人(Frank Cichocki博士)将使用高通量测序方法研究CMV诱导的适应性NK细胞的全球表观遗传学重构，并将确定驱动其增殖和分化的因素。Cichocki博士已经召集了一个由明尼苏达大学共济会癌症中心(MMC)和外部机构的专家组成的团队来指导拟议的研究活动，并在他向独立的过渡期间提供指导。除了来自导师和咨询委员会的直接支持外，奇科基博士还将完全访问MMC中可用的共享资源。其中包括流式细胞术、高通量测序和基因组学、生物统计学和信息学方面的核心设施。Cichocki博士将在他的指导岗位上继续工作两年，同时分析CMV诱导的适应性NK细胞的表观遗传重塑；这项工作将使他在高通量测序技术方面获得进一步的专业知识，这将是 他未来研究事业的成功。随着Cichocki博士进入其职业生涯的独立阶段，他将专注于假设驱动的实验，这些实验基于初步数据，表明IL-15、IL-21、IL-12和CD16受体的结合推动了CMV诱导的适应性NK细胞的扩张。拟议研究所产生的数据将构成3.5-4年前完成的R01申请的基础。建议的研究具有重要意义，因为它们解决了基础研究中的一个相当大的空白，在它们可以用于临床之前，需要充分描述适应性NK细胞亚群的特征，并了解驱动它们分化和增殖的机制。这些发现可能对设计激发NK细胞记忆的CMV疫苗以及在造血细胞移植的背景下具有重要的临床意义，在造血细胞移植中，CMV的重新激活与防止白血病复发相关。奇科基博士的长期目标是，作为一家学术机构的独立研究员，致力于推动基础和翻译NK细胞生物学的发展。作为一名对分子生物学和表观遗传学有着浓厚兴趣和成熟记录的免疫学家，他在回答这一建议中提出的问题并迅速推进对NK细胞记忆的机制理解方面具有独特的地位。

 DESCRIPTION (provided by applicant): Project Summary Recent evidence suggests that natural killer (NK) cells can develop immunological memory against viral antigens. Our extensive preliminary data show that cytomegalovirus (CMV) infection is associated with the generation of novel populations of highly differentiated, adaptive NK cells. These NK cell subsets display a genome-wide DNA methylation profile that mirrors that of effector CD8+ T cells. In the present proposal, the applicant (Dr. Frank Cichocki) will study global epigenetic remodeling in CMV-induced adaptive NK cells using a high-throughput sequencing approach and will identify the factors that drive their proliferation and differentiation. Dr. Cichocki has assembled a team of experts from the University of Minnesota's Masonic Cancer Center (MMC) and outside institutions to guide the proposed research activities and provide mentorship during his transition to independence. In addition to direct support from mentors and an advisory committee, Dr. Cichocki will have full access to the shared resources available in the MMC. These include core facilities in flow cytometry, high-throughput sequencing and genomics, biostatistics, and informatics. Dr. Cichocki will continue in his mentored position for two years while analyzing epigenetic remodeling in CMV- induced adaptive NK cells; this work will allow him to gain further expertise in high-throughput sequencing technologies, which will be critical to the success of his future research career. As Dr. Cichocki transitions into the independent phase of his career, he will focus on hypothesis-driven experiments that are based on preliminary data showing that the combination of IL-15, IL-21, IL-12, and CD16 receptor engagement drive the expansion of CMV-induced adaptive NK cells. The data generated from the proposed studies will form the basis for an R01 application to be completed by year 3.5-4. The proposed studies are significant because they address a considerable gap in basic research that is needed to fully characterize adaptive NK cell subsets and understand the mechanisms that drive their differentiation and proliferation before they can be utilized in clinicl settings. These findings may have major clinical implications for the design of CMV vaccines that elicit NK cell memory and in the context of hematopoietic cell transplantation where CMV reactivation is associated with protection from leukemia relapse. Dr. Cichocki's long-term goal is to dedicate his career to advancing basic and translational NK cell biology as an independent investigator at an academic institution. As an immunologist with a deep interest and proven track record in molecular biology and epigenetics, he is in a unique position to answer the questions set forth in this proposal and to rapidly advance the mechanistic understanding of NK cell memory.