Role of Natural Killer Cell Diversity in Multiple Sclerosis Risk and Disease Course

项目摘要 该项目的目标是全面描述多形性自然杀伤细胞的作用 多发性硬化症的受体。大量数据表明NK细胞参与了MS的发病机制，但其确切的 在调节风险和疾病过程中的作用还知之甚少。考虑到大量和多样化的高度 可变受体控制着NK细胞的活性，它们在多发性硬化症中的特征有望产生重要的见解 自然杀伤细胞在疾病中的作用。因此，我们将描述基因组联合的性质和程度。 NK受体在一组不同的、已建立的和深表型多发性硬化症队列中的变异。这一目标将 通过我们新颖的高通量、高分辨率的下一代测序(NGS)方法实现。 使用最先进的技术，我们将通过考虑NK细胞的表型来了解这种基因组变异 疾病。最后，我们将探索观察到的NK受体变异的功能含义，允许 从机制上解释NK受体表达的影响，以更全面地了解这些高度 MS中的可变受体一起，19号染色体上的白细胞受体复合体(LRC)和天然的 12号染色体上的杀伤复合体(NKC)编码90多种不同的NK细胞受体，包括极端的 KIR(杀伤细胞免疫球蛋白样受体)和LILR(白细胞免疫球蛋白样受体)的多态。 在这里描述的工作中，我们通过几个关键模式处理这些复杂系统，检查 首次全面描述多发性硬化患者NK受体的基因组、表型和功能变异 NK受体在疾病中的作用。我们在一个欧洲血统队列中的初步工作发现了一种 KIR变异与多发性硬化症的发病风险以及临床和MRI特征显著相关 疾病。在具体目标1中，我们将把这些发现扩展到不同的患者群体，并描述KIR的特征 在具有纵向临床和MRI数据的额外队列中，多发性硬化症在不同祖先之间的变异，以及 研究所有已知的多态NK受体及其相应配体的基因组变异。在……里面 具体目标2，我们将通过检查颞叶来更充分地解决这些受体在疾病过程中的作用 纵向CyTOF分析MS患者NK细胞受体的表达模式最后，在 具体目标3，我们将通过利用CRISPR技术来表征影响，从而将这些结果与背景联系起来 NK细胞受体表达水平对NK细胞功能的影响。这项工作有望填补我们在 了解NK细胞在多发性硬化症中的作用，因为NK细胞的活性受到高度变量的严格控制 我们在这项提案中调查的受体，这项工作将为调节 NK细胞在MS易感性和病程中的作用机制。结果将会产生 关于我们对疾病发病机制的理解和未来开发以NK细胞为基础的治疗方法的努力 选择。

Project Summary The goal of this project is to comprehensively characterize the role of polymorphic natural killer (NK) cell receptors in multiple sclerosis (MS). Substantial data implicate NK cells in MS pathogenesis, but their precise function in mediating risk and disease course is poorly understood. Given that a large and diverse set of highly variable receptors govern NK cell activity, their characterization in MS promises to yield important insights into the role of NK cells in disease. Thus, we will characterize the nature and extent of the association of genomic variation of NK receptors across a set of diverse, established and deeply phenotyped MS cohorts. This goal will be achieved via our novel high-throughput, high-resolution next-generation sequencing (NGS) methodology. Using state-of-art technologies, we will contextualize this genomic variation by considering NK cell phenotype in disease. Finally, we will explore the functional implications of the observed NK receptor variation, allowing a mechanistic explanation of the impact of NK receptor expression to more fully understand the role of these highly variable receptors in MS. Together, the leukocyte receptor complex (LRC) on chromosome 19 and the natural killer complex (NKC) on chromosome 12 encode more than 90 distinct NK cell receptors, including the extremely polymorphic KIR (killer-cell immunoglobulin-like receptor) and LILR (leukocyte immunoglobulin-like receptors). In the work described here, we approach these complex systems across several key modalities, examining genomic, phenotypic and functional variation of NK receptors in MS to provide the first comprehensive depiction of the role of NK receptors in disease. Our preliminary work in a cohort of European ancestry identified a significant association of KIR variation with risk for developing MS, as well as with clinical and MRI features of disease. In Specific Aim 1, we will extend these findings to diverse patient populations and characterize KIR variation in MS across ancestries, in an additional cohort with longitudinal clinical and MRI data, and by investigating genomic variation of all known polymorphic NK receptors and their corresponding ligands. In Specific Aim 2, we will more fully resolve the role of these receptors in disease course by examining temporal expression patterns of NK cell receptors in individuals with MS through longitudinal CyTOF analysis. Finally, in Specific Aim 3, we will contextualize these results by harnessing CRISPR technology to characterize the impact of NK cell receptor expression level on NK cell function. This work promises to fill critical gaps in our understanding of the role of NK cells in MS. Because NK cell activity is tightly governed by the highly variable receptors that we investigate in this proposal, this work will provide important insight into the regulatory mechanisms underlying the influence of NK cells in MS susceptibility and disease course. The results will bear both on our understanding of disease pathogenesis and future efforts to develop NK cell-based therapeutic options.