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

项目摘要 本项目的目标是全面表征多态性自然杀伤（NK）细胞的作用， 多发性硬化症（MS）中的受体。大量数据表明NK细胞参与MS发病机制，但其精确的 在介导风险和疾病过程中的功能知之甚少。鉴于一个庞大而多样化的高度 可变受体控制NK细胞活性，它们在MS中的特征有望产生重要的见解， NK细胞在疾病中的作用因此，我们将描述基因组关联的性质和程度， NK受体在一组不同的、已建立的和深度表型化的MS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细胞表型来了解这种基因组变异， 疾病最后，我们将探讨观察到的NK受体变异的功能意义， 机制解释NK受体表达的影响，以更充分地了解这些高度 总之，19号染色体上的白细胞受体复合物（LRC）和MS中的天然白细胞受体复合物（LRC）。 12号染色体上的杀伤复合物（NKC）编码90多种不同的NK细胞受体，包括极端的 多态性KIR（白细胞免疫球蛋白样受体）和LILR（白细胞免疫球蛋白样受体）。 在这里描述的工作中，我们通过几种关键模式来处理这些复杂的系统， MS中NK受体的基因组、表型和功能变异，以提供第一个全面的描述 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确定了一个 KIR变异与MS发生风险以及MS的临床和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人群，并描述KIR的特征。 在具有纵向临床和MRI数据的额外队列中， 研究所有已知的多态性NK受体及其相应配体的基因组变异。在 具体目标2，我们将更充分地解决这些受体在疾病过程中的作用，通过检查时间 通过纵向CyTOF分析MS个体中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技术来描述这些结果的影响， 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的关键空白， 由于NK细胞活性受到高度可变的 受体，我们在这项建议中调查，这项工作将提供重要的洞察力的调节 NK细胞在MS易感性和疾病过程中的影响机制。结果将承担 无论是我们对疾病发病机制的理解，还是未来开发基于NK细胞的治疗药物的努力， 选项.

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.